Available online at www.sciencedirect.

com

Journal of Ethnopharmacology 115 (2008) 173183

Review

Animal models to test drugs with potential antidiabetic activity

T.S. Fr ode a, , Y.S. Medeiros b
Department of Clinical Analysis, Center of Health Sciences, Federal University of Santa Catarina, Campus Universit ario, Trindade, 88040-970, Florian opolis, SC, Brazil b Department of Pharmacology, Center of Biological Sciences, Federal University of Santa Catarina, Campus Universit ario, Trindade, 88049-900, Florian opolis, SC, Brazil Received 1 March 2007; received in revised form 26 October 2007; accepted 26 October 2007 Available online 4 November 2007
a

Abstract Although medicinal plants have been historically used for diabetes treatment throughout the world, few of them have been validated by scientic criteria. Recently, a large diversity of animal models has been developed to better understand the pathogenesis of diabetes mellitus and new drugs have been introduced in the market to treat this disease. The aim of this work was to review the available animal models of diabetes and some in vitro models which have been used as tools to investigate the mechanism of action of drugs with potential antidiabetic properties. In addition, a MEDLINE/PUBMED search for articles on natural products, pancreatectomy and diabetes mellitus treatment published between 1996 and 2006 was done. In the majority of the studies, natural products mainly derived from plants have been tested in diabetes models induced by chemical agents. This review contributes to the researcher in the ethnopharmacology eld to designs new strategies for the development of novel drugs to treat this serious condition that constitutes a global public health. 2007 Elsevier Ireland Ltd. All rights reserved.
Keywords: Diabetes mellitus; Animal models; Genetic studies; Natural products

Contents
1. 2. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . In vivo animal models of diabetes mellitus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1. Pharmacological induction of diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.2. Surgical models of diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3. Genetic models of diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3.1. Animal strains that spontaneously develop diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.3.2. Genetically engineered diabetic mice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.4. Other models of type 2 diabetes to evaluate the reduction of pancreatic -cell mass . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . In vitro studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1. In vitro studies on insulin secretion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1.1. Studies using isolated pancreatic islet cell lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1.2. Studies using insulin-secreting cell lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.2. In vitro studies on glucose uptake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174 174 174 178 178 178 178 179 179 179 179 179 179

3.

Abbreviations: ADP, adenosine diphosphate; AE, aqueous extract; AT, acute treatment; ATP, adenosine triphosphate; BB rat, Bio Breeding Laboratories rat; BuOH, n-butanol fraction; Ca2+ , calcium; CH2 Cl2 , methylene chloride extract; CH3 Cl, chloroform extract; db/db mouse, monogenic model of obesity (leptin resistant); DNA, deoxyribonucleic acid; DPP-4, dipeptidyl peptidase-4; EtOAc, ethyl acetate fraction; EtOH, ethanolic extract; GK rat, Goto-Kazikasi rat; GLUT, glucose transporter; Hex, hexane fraction; i.p., intraperitoneal route; i.v., intravenous route; IGFs, insulin-like growth factors; MeOH, methanolic extract; NOD mouse, non-obese diabetic mouse; Ob/Ob mouse, monogenic model of obesity mouse (leptin decient); OLETL rat, Otsuka Long-Evans Tokushima Fatty rat; p.o., oral route; STZ, streptozotocin; ZK rat, Zucker rat. Corresponding author. Tel.: +55 48 99614846; fax: +55 48 32440936. E-mail addresses: saleh@ccs.ufsc.br, taniafrode@zipmail.com.br (T.S. Fr ode). 0378-8741/$ see front matter 2007 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2007.10.038

174

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183

4. 5. 6.

Models of diabetes accelerated atherosclerosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Potential effects of novel antidiabetic drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Concluding remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

179 180 180 180

1. Introduction The increasing worldwide incidence of diabetes mellitus in adults constitutes a global public health burden. It is predicted that by 2030, India, China and the United States will have the largest number of people with diabetes (Wild et al., 2004). By denition, diabetes mellitus is categorized as a metabolic disease characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. The vast majority of cases of diabetes fall into two broad etiopathogenetic categories. In one category, type 1 diabetes, the cause is an absolute deciency of insulin secretion. In the other, much more prevalent category, type 2 diabetes, the cause is a combination of resistance to insulin action and an inadequate compensatory insulin-secretory response (American Diabetes Association, 2005). Despite the great interest in the development of new drugs to prevent the burden of complications associated with this disease and the raised interest in the scientic community to evaluate either raw or isolated natural products in experimental studies (Fig. 1), few of them were tested in humans (Liu et al., 2004; Vuksan and Sievenpiper, 2005; Johnson et al., 2006; Jung et al., 2006; Matsui et al., 2006). Nevertheless, natural supplements are widely used around the world to treat diabetes, but medical research does not support their effectiveness. In these studies the low methodological quality, small sample size of tested patients, and limited number of trials deserve caution in the interpretation of the positive data and require further examination in high-quality trials (Liu et al., 2004). Nowadays, clinical treatment of diabetes targets both insulin deciency and resistance and more recently the prevention of pancreatic -cell function decline (Hansotia and Drucker, 2005) (Table 1). The aim of this work was to review the available animal models of diabetes in order to provide adequate tools to investigate the mechanism of action of drugs with potential antidiabetic properties. Considering that there is a marked overlap between type 1 and 2 diabetes mellitus either in human beings or in ani-

mal models, some classical models for each type of this disease cannot also be sustained (Chaparro et al., 2006; Donath and Ehses, 2006). However, it is cited, when appropriated along the text, which category (type) the described model ts better. The information in this review was obtained by searching MEDLINE/PUBMED for literature published in English from 1996 to 2006, using terms natural products, pancreatectomy and diabetes mellitus treatment. The decision of which model of diabetes to use for any particular protocol is mainly inuenced by local resources. Ideally, preclinical experiments should be initially carried out in vivo, and be complemented, when possible, with in vitro studies to explore and advance in the mechanism of action of a natural product. 2. In vivo animal models of diabetes mellitus Diabetes can be induced by pharmacologic, surgical or genetic manipulations in several animal species. Most experiments in diabetes are carried out on rodents, although some studies are still performed in larger animals. The classical model employed by Banting and Best was pancreatectomy in dogs (Bliss, 2000). It is also described prone strains to diabetes mellitus that have been employed in several researches (Chen and Wang, 2005; Rees and Alcolado, 2005; Masiello, 2006). Currently, the murine model is one of the most used due to the availability of over 200 well-characterized inbred strains and the ability to delete or over-express specic genes through knockout and transgenic technologies (Rees and Alcolado, 2005; Masiello, 2006). 2.1. Pharmacological induction of diabetes The majority of studies published in the eld of ethnopharmacology between 1996 and 2006 employed these models. Streptozotocin (STZ, 69%) and alloxan (31%) are by far the most frequently used drugs and this model has been useful for the study of multiple aspects of the disease. Both drugs exert their diabetogenic action when they are administered parenterally: intravenously, intraperitoneally or subcutaneously. The dose of these agents required for inducing diabetes depends on the animal species, route of administration and nutritional status. According to the administered dose of these agents, syndromes similar to either type 1, type 2 diabetes mellitus or glucose intolerance can be induced (Lenzen et al., 1996; Mythili et al., 2004). Protocols are available any where, being critical the pH and type of buffer employed as well as the preparation of the solution of either alloxan or streptozotocin in the day of the experiments (Yu et al., 2000; Gupta et al., 2005a,b; Lei et al., 2005; Babu et al., 2006; Miranda et al., 2006).

Fig. 1. Number of papers that tested natural products in experimental models of diabetes published in the period of 19962006 (source: PUBMED, English language, accessed in 06-11-2007).

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183 Table 1 Drugs available to treat diabetes mellitus Insulin deciency Insulin and its analogs Sulphonylureas, glinides Insulin resistance Metformin Glitazones Alpha-glucosidase inhibitors

175

Prevention of -cell decline Amylin analogs Exenatide, DPP-4 inhibitors

The cytotoxic action of these diabetogenic agents is mediated by reactive oxygen species, but both drugs differ in their mechanism of action (Federiuk et al., 2004; Lei et al., 2005). Alloxan and the product of its reduction, dialuric acid, establish a redox cycle with the formation of superoxide radicals. These radicals undergo dismutation to hydrogen peroxide with a simultaneous massive increase in cytosolic calcium concentration, which causes rapid destruction of pancreatic -cells (Szudelski, 2001). The range of the diabetogenic dose of alloxan is quite narrow and even light overdosing may be generally toxic and may cause the loss of many animals. This loss is likely to stem from kidney tubular cell necrotic toxicity, in particular when too high doses of alloxan are administered (Lenzen et al., 1996). The most frequently used intravenous dose of alloxan in rats is 65 mg/kg, but when it is administered intraperitoneally (i.p.) or subcutaneously its effective dose must be higher (Federiuk et al., 2004). For instance, an intraperitoneal dose below 150 mg/kg may be insufcient for inducing diabetes in this animal species (Katsumata et al., 1992). In mice, doses vary among 100200 mg/kg by intravenous route (i.v.) (Machado et al., 2001; Miranda et al., 2006). Streptozotocin enters the pancreatic -cell via a glucose transporter-GLUT2 and causes alkylation of deoxyribonucleic acid (DNA). Furthermore, STZ induces activation of poly adenosine diphosphate ribosylation and nitric oxide release. As a result of STZ action, pancreatic -cells are destroyed by necrosis (Mythili et al., 2004). In adult rats, 60 mg/kg is the most common dose of STZ to induce insulin dependent diabetes (Patel et al., 2006), but higher doses are also used. STZ is also efcacious after intraperitoneal administration of a similar or higher dose, but single doses below 40 mg/kg may be ineffective (Katsumata et al., 1992). In general, rats are considered diabetic if tail blood glucose concentrations in fed animals are greater than 200300 mg/dl, 2 days after STZ injection. A model of type 2 diabetes can be induced in rats by either i.v. (tail vein) or i.p. treatment with STZ in the rst days of life. At 810 weeks of age and thereafter, rats neonatally treated with STZ manifest mild basal hyperglycemia, an impaired response to the glucose tolerance test, and a loss of pancreatic -cell sensitivity to glucose (Pascoe and Storlien, 1990). It has been observed that STZ at rst abolished the pancreatic -cell response to glucose, but a temporary return of responsiveness then appears which is followed by its permanent loss (Mythili et al., 2004). In adult mice, STZ given in multiple low doses (40 mg/kg, i.v. for 5 days) (Rees and Alcolado, 2005) induces an insulin dependent diabetes that is quite similar to the autoimmune forms (islet inammation and -cell death) of type 1 diabetes. On the other hand, a single dose between 60 and 100 mg/kg of STZ (Lei et al., 2005; Sharma et al., 2006), administered systemically can also cause insulin

dependent diabetes, but it lacks the autoimmune prole (Yu et al., 2000). The potential problem with STZ is that its toxic effects are not restricted to pancreatic -cells since it may cause renal injury (Valentovic et al., 2006), oxidative stress inammation and endothelial dysfunction (Lei et al., 2005). The destruction of pancreatic -cells by both drugs is associated with a huge release of insulin which makes animals more susceptible to severe hypoglycemia that may be lethal. Thus, following treatment with either STZ or alloxan, animals are fed with glucose solution (5%) for 1224 h. Afterwards, an increase of glucose levels is observed in comparison to control animals due to insulin deciency. It is also reported that fasted animals are more susceptible to alloxan effects (Katsumata et al., 1992; Federiuk et al., 2004) and increased blood glucose in fed animals provides partial protection (Federiuk et al., 2004). In general, experimental protocols recommend that administration of either STZ or alloxan must be done in the fasting period (812 h), followed by addition of glucose solution to avoid hypoglycemia. Besides rats, dogs and mice, other animal species such as rabbits and monkeys have been employed to induce diabetes by these protocols, but rabbits and pigs are more resistant to STZ (Rees and Alcolado, 2005). In general, by using these models of diabetes induced by chemical drugs, the majority of published studies report the amount of reduction of blood glucose that is always evaluated after a period of fasting following acute or chronic treatment with a specic natural product. Comparative studies are carried out with nondiabetic and/or diabetic animal groups treated with known antidiabetic drugs, but results do not permit to further explore the mechanism of action of the studied natural product. Glucose is measured by standard glucose-oxidase or dehydrogenase assays, mainly by means of commercial meters available everywhere. Insulin determination is available in experimental animals by different methodologies (radioimmunoassayRIA or immunometric assays) (Esmaeili and Yazdanparast, 2004). In chronic experiments, A1c glycated hemoglobin can be also measured (Gupta et al., 2005b; Sathishsekar and Subramanian, 2005; Sekar et al., 2005; Shirwaikar et al., 2005; Narendhirakannan et al., 2006; Santhakumari et al., 2006; Tanaka et al., 2006). Table 2 displays a list of plants and/or their active compounds tested in diabetic animals (induced by either STZ or alloxan) published in the past 2 years. It is necessary to reemphasize that natural products display several effects besides lowering blood glucose in these experimental models. In view of the lack of parallel studies of their toxicity, these models of diabetes induced by either alloxan or STZ are considered a screening step in the search for drugs for the treatment of diabetes (Kecskemeti et al., 2002).

176

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183

Table 2 List of studied natural products with putative antidiabetic effects tested in either in vivo or in vitro models in the period of 20052006 (source: PUBMED, English language) Plant (family) Aegle marmelos (Rutaceae) Material EtOH/leaves Treatment i.p., 14 d Drug-induced diabetes STZ-rat Results Glucose, glycosylated hemoglobin, C-peptide, glucose tolerance,glycogen, insulin Glucose Glucose, insulin Glycosylated hemoglobin A1c Glucose, lipids Gluose; insulin Glucose, lipids, lipidic peroxidation, glucose tolerance Glycosylated hemoglobin, glucose tolerance Glucose, lipids Glucose Glucose Glucose Glucose Glucose, glycosylated hemoglobin, glycogen, lipids, oxidative stress Glucose, lipid peroxidation, insulin Glucose, lipid, glucose tolerance Glucose, lipids, insulin Glucose tolerance, glucose, oxidative stress Glucose Glucose Glucose Glucose Glucose, oxidative stress, GLUT4, insulin Glucose Glucose Glucose Glucose, glycosylated hemoglobin, oxidative stress, glycogen References Narendhirakannan et al. (2006)

AE/seeds AE/fruits Aloe vera (Liliaceae) EtOH, isolated compounds/leaves EtOH/leaves Whole plant, oil fraction AE, EtOH/leaves

p.o., 14 d p.o., 30 d p.o., 28 d p.o., 21 d p.o., 21 d p.o., 1030 d

STZ-rat STZ-rat db/db mice STZ-rat STZ-rat STZ-rat

Kesari et al. (2006) Kamalakkannan and Prince (2005) Tanaka et al. (2006) Rajasekaran et al. (2004) Kim et al. (2006) Gupta et al. (2005a); Kaleem et al. (2006)

Amaranthus esculantus (Amaranthaceae) Annona squamosa (Annonaceae)

Fruit-pulp Averrhoa bilimbi (Oxalidaceae) Baccharis trimera (Myrtaceae) Bryophyllum pinnatum (Crassulaceae) Canarium schweinfurthi (Burseraceae) Chamaemelum nobile (Asteraceae) Coscinium fenestratum (Menispermaceae) Egyptian Morus alba (Moraceae) Eugenia jambolana (Asteraceae) Garlic (Allium sativum L.) (Liliaceae) AE, BuOH, EtOAc, Hex/leaves AE, EtOH, BuOH/leaves AE/leaves MeOH/CH2 Cl2 /stem barks AE/leaves AAE/stem barks

p.o., 10 d p.o., 15 d p.o., 14 d p.o., 7 d p.o./i.p., AT p.o.,14 d p.o., 15 d p.o., 12 d

Alloxan-rabbit STZ-rat, alloxan-rabbit STZ-rat STZ-mice STZ-rat STZ-rat STZ-rat STZ-rat

Gupta et al. (2005b) Tan et al. (2005) Oliveira et al. (2005) Ojewole (2005a) Kamtchouing et al. (2005) Eddouks et al. (2005a) Shirwaikar et al. (2005)

AAE, isolated compounds/stem barks AE, EtOH, isolated compounds/fruitpulp/seeds EtOH/bulbs Garlic oil, isolated compounds

p.o., 10 d

STZ-rat

Singap et al. (2005)

p.o., AT

STZ-rabbit

Sharma et al. (2006); Ravi et al. (2005) Eidi et al. (2006) Liu et al. (2005, 2006); El-Demerdash et al. (2005) Guerrero-Analco et al. (2005); Navarrete and Mata (2005) Ojewole (2006) Ojewole (2005b) Eddouks et al. (2005c) Zhao et al. (2005b); Wu et al. (2006) Andrade-Cetto et al. (2005) Ojewole (2005c) Shetty et al. (2005); Harinantenaina et al. (2006) Sekar et al. (2005); Reyes et al. (2006)

p.o., 14 d p.o., 21112 d p.o., AT

STZ-rat STZ-rat alloxan STZ-rat

Hintonia standleyan (Rubiaceae) Hypoxis hemerocallidea (Hypoxidaceae) Leonotis leonurus (Lamiaceae) Lepidium sativum (Brassicaceae) Lycium barbarum (Solanaceae) Malmea depressa (Annonaceae) Mangifera indica (Anacardiaceae) Momordica charantia (Cucurbitaceae)

MeOH, isolated compounds/stem barks AE/fruits AE/leaves AE/leaves Isolated compound/fruits AE, EtOH, BuOH/roots AE/stem barks MeOH, isolated compounds/gourd AE/leaves

p.o., AT p.o., AT p.o., AT, p.o.,15 d p.o., 2126 d p.o., AT i.p., AT p.o., AT p.o., 2730 d

STZ-mice, rat STZ-mice, rat STZ-rat STZ-rat STZ-rat SZT-rat SZT-mice SZT-rat STZ-rat, alloxan-rat

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183 Table 2 (Continued ) Plant (family) Material AE/seeds Treatment p.o., 30 d Drug-induced diabetes STZ-rat Results Glucose, glycosylated hemoglobin, oxidative stress, lipid peroxidation Glucose, lipids Glucose, glycosylated hemoglobin Glucose Glucose, lipids, insulin Glucose Glucose gluconeogenesis Glucose, lipids, 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase activity, oxidative stress, insulin Glucose Glucose, hepatic glycogen Glucose Glucose Glucose Glucose Glucose Glucose Glucose Glucose, lipidic peroxidation Glucose, insulin References Sathishsekar and Subramanian (2005) Yadav et al. (2005)

177

Fruits Piper betle (Piperaceae) AE, EtOH/leaves

p.o.,21 d p.o., 30 d p.o., AT

Alloxan-rat STZ-rat

Arambewela et al. (2005); Santhakumari et al. (2006) Ojewole (2005d) Taniguchi et al. (2006) Maghrani et al. (2005) Lima et al. (2006) Latha and Pari (2005); Pari and Latha (2005, 2006)

Psidium guajava Linn. (Myrtaceae) Raphanus sativus (Brassicaceae) Retama raetam (Fabaceae) Salvia ofcinalis (Lamiaceae) Scoparia dulcis (Scrophulariacae)

AE/leaves AE/whole plant AE/whole plant AE/leaves AE/whole plant

p.o., AT p.o., 21 d i.v., AT p.o., 14 d p.o., 2142 d

STZ-rat STZ-rat STZ-rat STZ-rat, mice STZ-rat-in vitro/in vivo

Strobilanthes crispus (Acanthaceae) Syzygium cordatum (Myrtaceae) Syzygium cumini (synonym Tamarindus indica (Caesalpiniaceae) Taxus yunnanensis (Taxaceae) Terminalia chebula (Combretaceae) Terminalia superba (Combretaceae) Trema orientalis (Ulmaceae) Tremella mesenterica (Combretaceae) Triticum repens P. Beauv. (Gramineae) Viscum album L. (Loranthaceae) Zizyphus spina-christi (Rhamnaceae)

AE/leaves AE/leaves AE, EtOH, BuOH fraction/leaves and seeds AE, MeOH, isolated compounds/woods CH3 Cl extract/seeds MeOH, CH2 CL2 /stem barks AE/stem barks Isolated compounds/fruits AE/rhizomes AE, EtOH/whole plant BuOH, isolated compounds/leaves

p.o., 21 d p.o., 28 d p.o., 714 d

STZ-rat STZ-rat STZ-mice, rat

Fadzelly et al. (2006) Musabayane et al. (2005) Maiti et al. (2005); Oliveira et al. (2005) Banskota et al. (2006) Rao and Nammi (2006) Kamtchouing et al. (2005) Dimo et al. (2006) Lo et al. (2006) Eddouks et al. (2005b) Orhan et al. (2005) Abdel-Zaher et al. (2005)

i.p., AT p.o., AT p.o., 14 d p.o., AT p.o., 14 d p.o., AT p.o., 7 d p.o., AT

STZ-rat STZ-rat STZ-rat STZ-rat STZ-rat STZ-rat STZ-rat STZ-rat

AAE: Alcoholic extract, AE: aqueous extract, AT: acute treatment, BuOH: n-butanol fraction, CH2 Cl2 : methylene chloride extract, CH3 Cl: chloroform extract, EtOAc: ethyl acetate fraction, EtOH: ethanolic extract, GLUT-4: glucose transporter, Hex: hexane fraction, i.p.: intraperitoneal route, MeOH: methanolic extract, p.o.: oral route, and STZ: streptozotocin.

In addition, considering the diversity of active principles found in the crude plant extracts some studies also include further analysis of lipids (cholesterol, and triglycerides) as well as additional evaluation of the antioxidant properties of the product. This can be exemplied with studies with the fruit-pulp, seeds and leaves extracted from Eugenia jambolona, whose antidiabetic properties are largely recognized by folk medicine in several countries (Grover et al., 2002). Despite of the lack of dose standardization, plant environment selection and toxicological studies, different studies have conrmed that the ethanolic extracts of either the fruit-pulp or seeds of this plant present hypoglycemic, hypolipemic and antioxidant properties (Sharma et al., 2003; Pepato et al., 2005). These results suggest that numerous active principles may be present in the raw extract of

the plant and highlight the need to further advance in their characterization. In line with this hypothesis, Tanaka et al. (2006) had isolated several compounds with antihyperglycemic properties from Aloe vera leaves (Tanaka et al., 2006). The identication of these products explains, in part, why these compounds present antioxidant, antihyperglycemic, antilypemic properties and even enhance the process of wound healing in diabetic and nondiabetic animals (Okyar et al., 2001; Sharma et al., 2003, 2006; Pepato et al., 2005; Rajasekaran et al., 2005; Tanaka et al., 2006). Due to the nonspecic action of compounds isolated from extracts of natural products, some studies have aggregated additional in vitro protocols to the in vivo studies such as liver perfusion to evaluate glucose inux inhibition (Chung et al., 2006), gastrointestinal absorption methodologies and antiox-

178

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183

idant enzymatic systems, liver glycogen level, among others (Babu et al., 2006; Fadzelly et al., 2006; McAnuff-Harding et al., 2006). These protocols contribute to extend the analysis of the antidiabetic effects of a certain natural product. In this context, liver perfusion methodologies with the simultaneous measurement of glucose inux help to elucidate if the natural product exerts extra-pancreatic effects like metformin and glitazones. On the other hand, other studies suggest that inhibition of carbohydrate absorption may be linked to the antidiabetic properties of the natural product. Thus, inclusion of at least two different routes of treatment, for instance, i.p. and oral route (p.o.) can help in the analysis of the possible site of action of a studied natural product. Vacor, dithizone(diphenylthiocarbazone), and 8hydroxyquinolone may also cause experimental diabetes, but their use in research is restricted due to their level of toxicity (Clark et al., 1994). 2.2. Surgical models of diabetes Another technique used to induce diabetes is the complete removal of the pancreas. Few researchers have employed this model in the last years to explore effects of natural products with animal species such as rats, pigs, dogs and primates (Choi et al., 2004; Liu et al., 2004; Rees and Alcolado, 2005; Masiello, 2006). Limitations to this technique include (1) high level of technical expertise and adequate surgical room environment, (2) major surgery and high risk of animal infection, (3) adequate post-operative analgesia and antibiotic administration, (4) supplementation with pancreatic enzymes to prevent malabsorption and (5) loss of pancreatic counter regulatory response to hypoglycemia. More recently, partial pancreatectomy has been employed, but large resection (more than 80% in rats) is required to obtain mild to moderate hyperglycemia. In this case, small additional resection can result in signicant hypoinsulinemia (Masiello, 2006). Choi et al. (2004) investigated the action or relative glucose uptake in various tissues of 90% pancreatectomized rats by using either hyperglycemic or euglycemic hyperinsulinemic clamp methodologies. This experimental design permits to evaluate if the compound has some effect upon both resistance to and secretion of insulin. 2.3. Genetic models of diabetes 2.3.1. Animal strains that spontaneously develop diabetes These models permit the evaluation of the effect of a natural product in an animal without the interference of side effects induced by chemical drugs like alloxan and STZ reported above. Several recent publications summarized the major advances in this eld (for review see Rees and Alcolado, 2005; Masiello, 2006) and at the website http://www.informatics.jax. org). Similar to the human condition, these strains display complex and heterogeneous characteristics. In some of these models, insulin resistance predominates in association with obesity, dyslipidemia and hypertension, which provides valuable insights to

Fig. 2. Number of papers that tested natural products in prone strains to diabetes published in the period of 19962006 (source: PUBMED, English language, accessed in 06-11-2007).

study some events that are observed in human type 2 diabetes mellitus. Conversely, some strains like Ob/Ob mouse may maintain euglycemia due to a robust and persistent compensatory pancreatic -cell response, matching the insulin resistance with hyperinsulinemia. On the other hand, the db/db mouse rapidly develops hyperglycemia since their pancreatic -cells are unable to maintain the high levels of insulin secretion required throughout life. Thus, food intake is important in determining the severity of the diabetic phenotype and restriction of energy intake reduces both the obesity and hyperglycemia seen in this strain of mice. Another example is the spontaneously diabetic Goto-Kakizaki rat which is a genetic lean model of type 2 diabetes originating from selective breeding over many generations of glucose-intolerant nondiabetic Wistar rats (Chen and Wang, 2005). Regarding type 1 diabetes models, the NOD mouse typically presents hyperglycemia between 12 and 30 weeks of age, whereas in BB rats it occurs around 12 weeks of age. One great advantage of these models is that they can also be employed as model of atherosclerosis which represents the long-term complication of diabetes mellitus and tested against several natural products (Wu and Huan, 2007). As it is shown in Fig. 2, the NOD mouse followed by the ZK rat are the most used to test natural products. Other prone strains to type 1 diabetes mellitus include New Zealand white rabbit, Kreesbond dog, Chinese hamster and Celebes black ape. However, they have not been employed in studies to evaluate natural products to treat diabetes, except in preclinical trials of exenatide (incretin analog) (Rees and Alcolado, 2005). 2.3.2. Genetically engineered diabetic mice In this case, rodents may be produced to over (transgenic) or under (knockout)-express proteins thought to play a key part in glucose metabolism. For a sound review on this topic see Masiello (2006) and Clee and Attie (2006). Although signicant advances in this eld have arisen in recent years, especially with the advent of transgenic mice, there have been no studies carried out involving natural products and these models. Certainly, the high costs restrict their study in sophisticated protocols which explore mechanisms of potential therapeutic agents that either stimulate pancreatic -cell growth or inhibit pancreatic -cell death (Meiton, 2006).

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183

179

2.4. Other models of type 2 diabetes to evaluate the reduction of pancreatic -cell mass The importance of the progressive loss of pancreatic -cell reduction in the course of type 2 diabetes has been the focus of therapeutic targets in the development of novel and potential drugs acting by enhancing pancreatic -cell growth and/or survival (Hansotia and Drucker, 2005; Masiello, 2006). Experimental reduction of pancreatic -cell mass by either surgical or chemical means has been reported above, but these models are limited due to the variety amount of residual pancreatic -cell content. On the other hand, experimental studies carried out predominantly in rodents have demonstrated that incretins, IGFs, hepatocyte growth factor, pituitary adenylate cyclase-activating polypeptide are capable of enhancing pancreatic -cell mass in the eld of experimental diabetes (Stoffers et al., 2003). Hence, prolonged administration of these agents may expand pancreatic -cell mass, leading to increased insulin secretion and improved glycemic control (Hansotia and Drucker, 2005). A method to induce diabetes in adult rats is to mimic the unfavorable intrauterine environment, which in humans leads to low-birth weight and is supposed to confer high risk for the development of diabetes in adult age (Holemans et al., 2003). This model known as intrauterine growth retardation by uteroplacental insufciency in the rat is based on the premise that uterine malnutrition may also increase the risk of diabetes amongst offspring in later life (Simmons et al., 2001). This has been achieved by several means, including bilateral uterine artery ligation (Simmons et al., 2001; Boloker et al., 2002), at 19 days of gestation, i.e. 3 days before term. The diabetogenic effects of manipulating the intrauterine environment are probably mediated by a permanent programming of the developing offspring, e.g. by the mechanism of imprinting. It should also be pointed out that the increased risk of diabetes continues into subsequent generations, which in turn, suggests that changes also affect the germ cell line (Reusens and Remacle, 2001). Finally, animal models with increased pancreatic -cell apoptosis have also been developed, besides those cited in Section 2.3 (Matveyenko and Butler, 2006). 3. In vitro studies 3.1. In vitro studies on insulin secretion Conventional antidiabetic agents can affect several pathways of glucose metabolism such as insulin secretion, glucose uptake by target organs as well as nutrient absorption. Recently, incretins (Hansotia and Drucker, 2005), and transcription factors such as peroxisome proliferator-activated receptorsPPAR (Rosenson, 2007) are targets of modern therapy. Insulin receptor, glucose transporters, however, has not been yet the focus of antidiabetic therapy. Although few studies using natural products have been published (Iwashima et al., 2001; Storling et al., 2005; Zhao et al., 2005a), these methodologies may serve as complementary tools to explore ndings obtained in in vivo models.

3.1.1. Studies using isolated pancreatic islet cell lines Several in vitro assays are available to study different steps of insulin secretion. It is known that insulin secretion occurs when pancreatic -cells utilize glucose to generate adenosine triphosphate (ATP) from adenosine diphosphate (ADP) (Affourtit and Brand, 2006). The resulting increase in cytoplasmic ATP/ADP ratio closes ATP-sensitive potassium channels, causing depolarization of the plasma membrane, which activates voltage-dependent Ca2+ channels. This results in elevation of the intracellular Ca2+ concentration which triggers insulin secretion (Ashcroft and Rorsman, 2004). In type 2 diabetes, pancreatic -cells exhibit atypical ion channel activity and an abnormal pattern of insulin secretion (Ashcroft and Rorsman, 2004). These pathways can be studied with isolated pancreatic -cells from either control or diabetic rat or mouse that can be obtained by collagenase digestion technique, followed by adequate separation and transference to appropriated culture medium (Zhao et al., 2005a; Storling et al., 2005). Afterwards, the experimental protocol is assayed. 3.1.2. Studies using insulin-secreting cell lines Bioengineered technologies have provided new opportunities to improve and establish more appropriate cultured cell lines to help to facilitate studies of mechanisms of both insulin secretion and -cell dysfunction, being also the target to the study of natural products. The most widely used insulin-secreting cell lines are RIN, HIT, beta-TC, MIN6 and INS-1 cells (Poitout et al., 1996). These cell lines release mainly insulin and small amounts of glucagon and somatostatin. Although the behaviour of none of these cell lines perfectly mimics primary -cell physiology, they are extremely valuable tools for the study of molecular events underlying -cell function (Poitout et al., 1996). 3.2. In vitro studies on glucose uptake Adipose tissue is considered a key link between obesity and type 2 diabetes by promoting the development of lipotoxicity, i.e. cell damage as a consequence of elevated intracellular lipid concentrations and insulin resistance (Lelliott and Vidal-Puig, 2004). Insulin resistance either at the adipocyte or skeletal muscle levels contribute to hyperglycemia. However, adipocytes from different sites of the body may have different biological or pathological effects. Pathways related to insulin resistance may be studied in cell lines of adipocytes such as murine 3T3-L1 cells (Karalee et al., 2001) and rat L6 muscle engineered to over-express GLUT4 (Maddux et al., 2001) and may be employed as tools to evaluate the effects of natural products upon glucose uptake. 4. Models of diabetes accelerated atherosclerosis Accelerated cardiovascular disease is a leading cause of both morbidity and mortality in diabetic patients (Wu and Huan, 2007). Aggressive therapy of dyslipidemia is necessary, since the risk of myocardial infarction is the same as in nondiabetic patients with previous myocardial infarction (Haffner et al., 1998). Currently, rats and mice are the most widely used mod-

180

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183 Babu, P.V., Sabitha, K.E., Shyamaladevi, C.S., 2006. Green tea impedes dyslipidemia, lipid peroxidation, protein glycation and ameliorates Ca2+ -ATPase and Na+ /K+ -ATPase activity in the heart of streptozotocin-diabetic rats. Chemico-Biological Interactions 162, 157164. Banskota, N.T., Nguyen, Y., Tezuka, T., Nobukawa, S., Kadota, S., 2006. Hypoglycemic effects of the wood of Taxus yunnanensis on streptozotocininduced diabetic rats and its active components. Phytomedicine 13, 109 114. Bliss, M., 2000. The Discovery of Insulin. University of Chicago Press, Chicago, USA, pp. 3211418. Boloker, J., Gertz, S.J., Simmons, R.A., 2002. Gestational diabetes leads to the development of diabetes in adulthood in the rat. Diabetes 51, 14991506. Breyer, M.D., Bottinger, E., Brosius, F.C., Coffman, T.M., Fogo, A., Harris, R.C., Heilig, C.W., Sharma, K., 2005. Diabetic nephropathy: of mice and men. Advances in Chronic Kidney Disease 12, 128145. Chaparro, R.J., Konigshofer, Y., Beilhack, G.F., Shizuru, J.A., McDevitt, H.O., Chien, Y.H., 2006. Nonobese diabetic mice express aspects of both type 1 and type 2 diabetes. Proceedings of the National Academy of Sciences of the United States of America 103, 1247512480. Chen, D., Wang, M.W., 2005. Development and application of rodent models for type 2 diabetes. Diabetes, Obesity and Metabolism 7, 307317. Choi, S.B., Park, C.H., Choi, M.K., Jun, D.W., Park, S., 2004. Improvement of insulin resistance and insulin secretion by water extracts of Cordyceps militaris, Phellinus linteus, and Paecilomyces tenuipes in 90% pancreatectomized rats. Bioscience, Biotechnology, and Biochemistry 68, 2257 2264. Chung, M.Y., Rho, M.C., Le, S.W., Park, H.R., Lee, I.A., Kim, D.H., Jeune, K.H., Lee, H.S., Kim, Y.K., 2006. Inhibition of diacylglycerol acyltransferase by betulinic acid from Alnus hirsute. Planta Medica 72, 267269. Clark, S.A., Borland, K.M., Sherman, S.D., Rusack, T.C., Chick, W.L., 1994. Staining and in vitro toxicity of dithizone with canine, porcine, and bovine islets. Cell Transplant 3, 299306. Clee, S.M., Attie, A.D., 2006. The genetic landscape of type-2 diabetes in mice. Endocrinology Reviews 28, 4883. Dimo, T., Ngueguim, F.T., Kamtchouing, P., Dongo, E., Tan, P.V., 2006. Glucose lowering efcacy of the aqueous stem bark extract of Trema orientalis (Linn) Blume in normal and streptozotocin diabetic rats. Die Pharmazie 61, 233236. Donath, M.Y., Ehses, J.A., 2006. Type 1, type 1.5, and type 2 diabetes: NOD the diabetes we thought it was. Proceedings of the National Academy of Sciences of the United States of America 103, 1221712218. Eddouks, M., Lemhadri, A., Zeggwagh, N.A., Michel, J.B., 2005a. Potent hypoglycaemic activity of the aqueous extract of Chamaemelum nobile in normal and streptozotocin-induced diabetic rats. Diabetes Research and Clinical Practices 67, 189195. Eddouks, M., Maghrani, M., Michel, J.B., 2005b. Hypoglycaemic effect of Triticum repens P. Beauv in normal and diabetic rats. Journal of Ethnopharmacology 102, 228232. Eddouks, M., Maghrani, M., Zeggwagh, N.A., Michel, J.B., 2005c. Study of the hypoglycaemic activity of Lepidium sativum L. aqueous extract in normal and diabetic rats. Journal of Ethnopharmacology 97, 391395. Eidi, A., Eidi, M., Esmaeili, E., 2006. Antidiabetic effect of garlic (Allium sativum L.) in normal and streptozotocin-induced diabetic rats. Phytomedicine: International Journal of Phytotherapy and Phytopharmacology 13, 624629. El-Demerdash, F.M., Yousef, M.I., El-Naga, N.I., 2005. Biochemical study on the hypoglycemic effects of onion and garlic in alloxan-induced diabetic rats. Food and Chemical Toxicology 43, 5763. Esmaeili, M.A., Yazdanparast, R., 2004. Hypoglycaemic effect of Teucrium polium: studies with rat pancreatic islets. Journal of Ethnopharmacology 95, 2730. Fadzelly, A.B., Asmah, R., Fauziah, O., 2006. Effects of Strobilanthes crispus tea aqueous extracts on glucose and lipid prole in normal and streptozotocininduced hyperglycemic rats. Plant Foods for Human Nutrition 61, 712. Federiuk, I.F., Casey, H.M., Quinn, M.J., Wood, M.D., Ward, W.K., 2004. Induction of type-1 diabetes mellitus in laboratory rats by use of alloxan: route of administration, pitfalls, and insulin treatment. Comparative Medicine 54, 252257.

els to study diabetes and atherosclerosis. Albeit, diabetic mice do not exhibit a high degree of atherosclerosis unless hyperglycemia is associated with severe hyperlipidemia, a fat diet is also present in these protocols. Models of diabetic nephropathy, a microvascular complication have also been developed (Breyer et al., 2005; Wu and Huan, 2007). For a review of these models and other informations see the website: www.mmpc.org. 5. Potential effects of novel antidiabetic drugs At present no drug is able to arrest the progressive loss of pancreatic -cells which occurs in type 2 diabetes mellitus. According to the United Kingdom Prospective Diabetes StudyUKPDS results, at the time of type 2 diabetes mellitus diagnoses, 50% of pancreatic -cell function had already been lost (Kan, 2001). Thus, the efcacy and side effect of marketed oral antidiabetic drugs still need to be optimized. The recent introduction in the market of incretin analogs opened a new eld to evaluate drugs with putative properties that may cause both proliferation and maturation of human pancreatic -cells (Hansotia and Drucker, 2005). Currently, a standard model of experimental diabetes to study effects of drugs, which could help in preventing the progressive loss of pancreatic islet function remains to be established. 6. Concluding remarks In spite of the worldwide use of herbs and medicinal plants, the effective treatment of diabetes with phytochemicals has not been validated with scientic criteria which may support their substitution for the current therapy. Although some studies have been published with raw natural products they have neither shed light on the mechanisms of action of these products nor have they shown a potential to be employed as new therapeutic drugs. This implies that several models are necessary to called for, in addition to the demonstration that a putative natural product exerts antihyperglycemic activity. Thus, by focusing on other targets of pancreatic islet cell dysfunction, new models may help to elucidate effects of medicinal plants employed in the treatment of diabetes mellitus. References
Abdel-Zaher, A.O., Salim, S.Y., Assaf, M.H., Abdel-Hady, R.H., 2005. Antidiabetic activity and toxicity of Zizyphus spina-christi leaves. Journal of Ethnopharmacology 101, 129138. Affourtit, C., Brand, M.D., 2006. Stronger control of ATP/ADP by proton leak in pancreatic beta-cells than skeletal muscle mitochondria. The Biochemical Journal 393, 151159. American Diabetes Association, 2005. Diagnosis and classication of diabetes mellitus. Diabetes Care 28, 3742. Andrade-Cetto, A., Martinez-Zurita, E., Wiedenfeld, H., 2005. Hypoglycemic effect of Malmea depressa root on streptozotocin diabetic rats. Journal of Ethnopharmacology 100, 319322. Arambewela, L.S., Arawwawala, L.D., Ratnasooriya, W.D., 2005. Antidiabetic activities of aqueous and ethanolic extracts of Piper betle leaves in rats. Journal of Ethnopharmacology 102, 239245. Ashcroft, F.M., Rorsman, P., 2004. Molecular defects in insulin secretion in type-2 diabetes. Reviews in Endocrine and Metabolic Disorders 5, 135142.

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183 Guerrero-Analco, J.A., Hersch-Martinez, P., Pedraza-Chaverri, J., Navarrete, A., Mata, R., 2005. Antihyperglycemic effect of constituents from Hintonia standleyana in streptozotocin-induced diabetic rats. Planta Medica 71, 10991105. Gupta, R.K., Kesari, A.N., Murthy, P.S., Chandra, R., Tandon, V., Watal, G., 2005a. Hypoglycemic and antidiabetic effect of ethanolic extract of leaves of Annona squamosa L. in experimental animals. Journal of Ethnopharmacology 99, 7581. Gupta, R.K., Kesari, A.N., Watal, G., Murthy, P.S., Chandra, R., Tandon, V., 2005b. Nutritional and hypoglycemic effect of fruit pulp of Annona squamosa in normal healthy and alloxan-induced diabetic rabbits. Annals of Nutrition and Metabolism 49, 407413. Grover, J.K., Yadav, S., Vats, V., 2002. Medicinal plants of India with antidiabetic potential. Journal of Ethnopharmacology 81, 81100. Haffner, S.M., Lehto, S., R onnemaa, T., Py or al a, K., Laakso, M., 1998. Mortality from coronary heart disease in subjects with type 2 diabetes and in nondiabetic subjects with and without prior myocardial infarction. New England Journal of Medicine 339, 229234. Hansotia, T., Drucker, D.J., 2005. GIP and GLP-1 as incretin hormones: lessons from single and double incretin receptor knockout mice. Regulatory Peptides 128, 125134. Harinantenaina, L., Tanaka, M., Takaoka, S., Oda, M., Mogami, O., Uchida, M., Asakawa, Y., 2006. Momordica charantia constituents and antidiabetic screening of the isolated major compounds. Chemical and Pharmaceutical Bulletin 54, 10171021. Holemans, K., Aerts, L., Van Assche, F.A., 2003. Fetal growth restriction and consequences for the offspring in animal models. Journal of the Society for Gynecologic Investigation 10, 392399. Iwashima, Y., Abiko, A., Ushikubi, F., Hata, A., Kaku, K., Sano, H., Eto, M., 2001. Downregulation of the voltage-dependent calcium channel (VDCC) -subunit mRNAs in pancreatic islets of type 2 diabetic rats. Biochemical and Biophysical Research Communications 280, 923932. Johnson, L., Strich, H., Taylor, A., Timmermann, B., Malone, D., Teufel-Shone, N., Drummond, R., Woosley, R., Pereira, E., Martinez, A., 2006. Use of herbal remedies by diabetic Hispanic women in the southwestern United States. Phytotherapy Research 20, 250255. Jung, M., Park, M., Lee, H.C., Kang, Y.H., Kang, E.S., Kim, S.K., 2006. Antidiabetic agents from medicinal plants. Current Medicinal Chemistry 13, 12031218. Kaleem, M., Asif, M., Ahmed, Q.U., Bano, B., 2006. Antidiabetic and antioxidant activity of Annona squamosa extract in streptozotocin-induced diabetic rats. Singapore Medical Journal 47, 670675. Kamalakkannan, N., Prince, P.S., 2005. The effect of Aegle marmelos fruit extract in streptozotocin diabetes: a histopathological study. Journal of Herbal Pharmacotherapy 5, 8796. Kamtchouing, P., Kahpui, S.M., Dzeuet, P.D., Tedong, L., Asongalem, E.A., Dimo, T., 2005. Anti-diabetic activity of methanol/methylene chloride stem bark extracts of Terminalia superba and Canarium schweinfurthii on streptozotocin-induced diabetic rats. Journal of Ethnopharmacology 104, 306309. Kan, E.E., 2001. The importance of -cell failure in the development and progression of type 2 diabetes. The Journal of Clinical Endocrinology and Molecular Metabolism 86, 40474058. Karalee, J.J., Anderson, R.A., Graves, D.A., 2001. Hydroxychalcone derived from cinnamon functions as a mimetic for insulin in 3T3-L1 adipocytes. Journal of the American College of Nutrition 20, 327336. Katsumata, K., Katsumata Jr., K., Katsumata, Y., 1992. Protective effect of diltiazem hydrochloride on the occurrence of alloxan- or streptozotocin-induced diabetes in rats. Hormone and Metabolic Research 24, 508510. Kecskemeti, V., Bagi, Z., Pacher, P., Posa, I., Kocsis, E., Koltai, M.Z., 2002. New trends in the development of oral antidiabetic drugs. Current Medicinal Chemistry 9, 5371. Kesari, A.N., Gupta, R.K., Singh, S.K., Diwakar, S., Watal, G., 2006. Hypoglycemic and antihyperglycemic activity of Aegle marmelos seed extract in normal and diabetic rats. Journal of Ethnopharmacology 107, 374379. Kim, H.K., Kim, M.J., Cho, H.Y., Kim, E.K., Shin, D.H., 2006. Antioxidative and anti-diabetic effects of amaranth (Amaranthus esculantus) in streptozotocininduced diabetic rats. Cell Biochemistry and Function 24, 195199.

181

Latha, M., Pari, L., 2005. Effect of an aqueous extract of Scoparia dulcis on plasma and tissue glycoproteins in streptozotocin induced diabetic rats. Die Pharmazie 60, 151154. Lei, Y.C., Hwang, J.S., Chan, C.C., Lee, C.T., Cheng, T.J., 2005. Enhanced oxidative stress and endothelial dysfunction in streptozotocin-diabetic rats exposed to ne particles. Environmental Research 99, 335343. Lenzen, S., Tiedge, M., Jorns, A., Munday, R., 1996. Alloxan derivatives as a tool for the elucidation of the mechanism of the diabetogenic action of alloxan. In: Shafrir, E. (Ed.), Lessons from Animal Diabetes. Birkhauser, Boston, pp. 113122. Lelliott, C., Vidal-Puig, A.J., 2004. Lipotoxicity, an imbalance between lipogenesis de novo and fatty acid oxidation. International Journal of Obesity and related Metabolic Disorders 28, 2228. Lima, C.F., Azevedo, M.F., Araujo, R., Fernandes-Ferreira, M., Pereira-Wilson, C., 2006. Metformin-like effect of Salvia ofcinalis (common sage): is it useful in diabetes prevention? The British Journal of Nutrition 96, 326-233. Liu, C.T., Hse, H., Lii, C.K., Chen, P.S., Sheen, L.Y., 2005. Effects of garlic oil and diallyl trisulde on glycemic control in diabetic rats. European Journal of Pharmacology 516, 165173. Liu, J.P., Zhang, M., Wang, W.Y., Grimsgaard, S., 2004. Chinese herbal medicines for type-2 diabetes mellitus. Cochrane Database of Systematic Reviews 3, 3642. Liu, C.T., Wong, P.L., Lii, C.K., Hse, H., Sheen, L.Y., 2006. Antidiabetic effect of garlic oil but not diallyl disulde in rats with streptozotocin-induced diabetes. Food and Chemical Toxicology 44, 13771384. Lo, H.C., Tsai, F.A., Wasser, S.P., Yang, J.G., Huang, B.M., 2006. Effects of ingested fruiting bodies, submerged culture biomass, and acidic polysaccharide lucuronoxylomannan of Tremella mesenterica Retz: Fr. on glycemic responses in normal and diabetic rats. Journal of Life Science 78, 1957 1966. Machado, A.F., Zimmerman, E.F., Hovland Jr., D.N., Weiss, R., Collins, M.D., 2001. Diabetic embryopathy in C57BL/6J mice. Altered fetal sex ratio and impact of the splotch allele. Diabetes 50, 11931199. Maddux, B.A., See, W., Lawrence Jr., J.C., Goldne, A.L., Goldne, I.D., Evans, J.L., 2001. Protection against oxidative stress-induced insulin resistance in rat L6 muscle cells by micromolar concentrations of alpha-lipoic acid. Diabetes 50, 404410. Maghrani, M., Michel, J.B., Eddouks, M., 2005. Hypoglycaemic activity of Retama raetam in rats. Phytotherapy Research 19, 125128. Maiti, R., Das, U.K., Ghosh, D., 2005. Attenuation of hyperglycemia and hyperlipidemia in streptozotocin-induced diabetic rats by aqueous extract of seed of Tamarindus indica. Biological and Pharmaceutical Bulletin 28, 11721176. Masiello, P., 2006. Animal models of type-2 diabetes with reduced pancreatic -cell mass. The International Journal of Biochemistry and Cell Biology 38, 873893. Matsui, T., Ogunwande, I.A., Abesundara, K.J.M., Matsumoto, K., 2006. Antihyperglycemic potential of natural products. Mini Reviews in Medicinal Chemistry 6, 109120. Matveyenko, A.V., Butler, P.C., 2006. Islet amyloid polypeptide (IAPP) transgenic rodents as models for type 2 diabetes. ILAR Journal/National Research Council, Institute of Laboratory Animal Resources 47, 225 233. McAnuff-Harding, M.A., Omoruyi, F.O., Asemota, H.N., 2006. Intestinal disaccharidases and some renal enzymes in streptozotocin-induced diabetic rats fed sapogenin extract from bitter yam (Dioscorea polygonoides). Life Sciences 78, 25952600. Meiton, D.A., 2006. Reversal of type 1 diabetes in mice. The New England Journal of Medicine 355, 8990. Miranda, M., Muriach, M., Roma, J., Bosch-Morell, F., Genoves, J.M., Barcia, J., Araiz, J., Diaz-Llospis, M., Romero, F.J., 2006. Oxidative stress in a model of experimental diabetic retinopathy: the utility of peroxynitrite scavengers. Archivos de la Sociedad Espa nola de Oftalmolog a 81, 2732. Musabayane, C.T., Mahlalela, N., Shode, F.O., Ojewole, J.A., 2005. Effects of Syzygium cordatum (Hochst.) [Myrtaceae] leaf extract on plasma glucose and hepatic glycogen in streptozotocin-induced diabetic rats. Journal of Ethnopharmacology 97, 485490.

182

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183 Ravi, K., Rajasekaran, S., Subramanian, S., 2005. Antihyperlipidemic effect of Eugenia jambolana seed kernel on streptozotocin-induced diabetes in rats. Food and Chemical Toxicology 43, 14331439. Rees, D.A., Alcolado, J.C., 2005. Animal models of diabetes mellitus. Diabetic Medicine 22, 359370. Reusens, B., Remacle, C., 2001. Intergenerational effect of an adverse intrauterine environment on perturbation of glucose metabolism. Twin Research 4, 406411. Reyes, B.A., Bautista, N.D., Tanquilut, N.C., Anunciado, R.V., Leung, A.B., Sanchez, G.C., Magtoto, R.L., Castronuevo, P., Tsukamura, H., Maeda, K.I., 2006. Anti-diabetic potentials of Momordica charantia and Andrographis paniculata and their effects on estrous cyclicity of alloxan-induced diabetic rats. Journal of Ethnopharmacology 105, 196200. Rosenson, R.S., 2007. Effects of peroxisome proliferator-activated receptors on lipoprotein metabolism and glucose control in type 2 diabetes mellitus. The American Journal of Cardiology 99, 96104. Santhakumari, P., Prakasam, A., Pugalendi, K.V., 2006. Antihyperglycemic activity of Piper betle leaf on streptozotocin-induced diabetic rats. Journal of Medicinal Food 9, 108112. Sathishsekar, D., Subramanian, S., 2005. Benecial effects of Momordica charantia seeds in the treatment of STZ-induced diabetes in experimental rats. Biological and Pharmaceutical Bulletin 28, 978983. Sekar, D.S., Sivagnanam, K., Subramanian, S., 2005. Antidiabetic activity of Momordica charantia seeds on streptozotocin induced diabetic rats. Pharmazie 60, 383387. Sharma, S.B., Nasir, A., Prabhu, K.M., Murthy, P.S., 2006. Antihyperglycemic effect of the fruit-pulp of Eugenia jambolana in experimental diabetes mellitus. Journal of Ethnopharmacology 104, 367373. Sharma, S.B., Nasir, A., Prabhu, K.M., Murthy, P.S., Dev, G., 2003. Hypoglycaemic and hypolipidemic effect of ethanolic extract of seeds of Eugenia jambolana in alloxan-induced diabetic rabbits. Journal of Ethnopharmacology 85, 201206. Shetty, A.K., Kumar, G.S., Sambaiah, K., Salimath, P.V., 2005. Effect of bitter gourd (Momordica charantia) on glycaemic status in streptozotocin induced diabetic rats. Plant Foods for Human Nutrition 60, 109 112. Shirwaikar, A., Rajendran, K., Punitha, I.S., 2005. Antidiabetic activity of alcoholic stem extract of Coscinium fenestratum in streptozotocin-nicotinamide induced type 2 diabetic rats. Journal of Ethnopharmacology 97, 369374. Simmons, R.A., Templeton, L.J., Gertz, S.J., 2001. Intrauterine growth retardation leads to the development of type 2 diabetes in the rat. Diabetes 50, 22792286. Singap, A.N., El-Beshbishy, H.A., Yonekawa, M., Nomura, T., Fukai, T., 2005. Hypoglycemic effect of Egyptian Morus alba root bark extract: effect on diabetes and lipid peroxidation of streptozotocin-induced diabetic rats. Journal of Ethnopharmacology 100, 333338. Stoffers, D.A., Desai, B.M., DeLeon, D.D., Simmons, R.A., 2003. Neonatal exendin-4 prevents the development of diabetes in the intrauterine growth retarded rat. Diabetes 52, 734740. Storling, J., Zaitsev, S.V., Kapelioukh, I.L., Karlsen, A.E., Billestrup, N., Berggren, P.O., Mandrup-Poulsen, T., 2005. Calcium has a permissive role in interleukin-1 -induced c-jun N-terminal kinase activation in insulinsecreting cells. Endocrinology 146, 30263036. Szudelski, T., 2001. The mechanism of alloxan and streptozotocin action in cells of the rat pancreas. Physiology Research 50, 536546. Tan, B.K., Tan, C.H., Pushparaj, P.N., 2005. Anti-diabetic activity of the semi-puried fractions of Averrhoa bilimbi in high fat diet fedstreptozotocin-induced diabetic rats. Journal of Life Science 76, 2827 2839. Tanaka, M., Misawa, E., Ito, Y., Habara, N., Nomaguchi, K., Yamada, M., Toida, T., Hayasawa, H., Takase, M., Inagaki, M., Higuchi, R., 2006. Identication of ve phytosterols from Aloe vera gel as antidiabetic compounds. Biological and Pharmaceutical Bulletin 29, 14181422. Taniguchi, H., Kobayashi-Hattori, K., Tenmyo, C., Kamei, T., Uda, Y., SugitaKonishi, Y., Oishi, Y., Takita, T., 2006. Effect of Japanese radish (Raphanus sativus) sprout (Kaiware-daikon) on carbohydrate and lipid metabolisms in normal and streptozotocin-induced diabetic rats. Phytotherapy Research 20, 274278.

Mythili, M.D., Vyas, R., Akila, G., Gunasekaran, S., 2004. Effect of streptozotocin on the ultrastructure of rat pancreatic islets. Microscopy Research and Technique 63, 274281. Narendhirakannan, R.T., Subramanian, S., Kandaswamy, M., 2006. Biochemical evaluation of antidiabetogenic properties of some commonly used Indian plants on streptozotocin-induced diabetes in experimental rats. Clinical and Experimental Pharmacology and Physiology 33, 11501157. Navarrete, A., Mata, R., 2005. Antihyperglycemic effect of constituents from Hintonia standleyana in streptozotocin-induced diabetic rats. Planta Medica 71, 10991105. Ojewole, J.A., 2005a. Antinociceptive, anti-inammatory and antidiabetic effects of Bryophyllum pinnatum (Crassulaceae) leaf aqueous extract. Journal of Ethnopharmacology 99, 1319. Ojewole, J.A., 2005b. Antinociceptive, antiinammatory and antidiabetic effects of Leonotis leonurus (L.) R. BR. [Lamiaceae] leaf aqueous extract in mice and rats. Methods and Findings in Experimental and Clinical Pharmacology 27, 257264. Ojewole, J.A., 2005c. Antiinammatory, analgesic and hypoglycemic effects of Mangifera indica Linn. (Anacardiaceae) stem-bark aqueous extract. Methods and Findings in Experimental and Clinical Pharmacology 27, 547554. Ojewole, J.A., 2005d. Hypoglycemic and hypotensive effects of Psidium guajava Linn. (Myrtaceae) leaf aqueous extract. Methods and Findings in Experimental and Clinical Pharmacology 27, 689695. Ojewole, J.A., 2006. Antinociceptive, anti-inammatory and antidiabetic properties of Hypoxis hemerocallidea Fisch. and C.A. Mey. (Hypoxidaceae) corm [African Potato] aqueous extract in mice and rats. Journal of Ethnopharmacology 103, 126134. Okyar, A., Can, A., Akev, N., Baktir, G., Sutlupinar, N., 2001. Effect of Aloe vera leaves on blood glucose level in type I and type II diabetic rat models. Phytotherapy Research 15, 157161. Oliveira, A.C., Endringer, D.C., Amorim, L.A., Das Grac as, L., Brandao, M., Coelho, M.M., 2005. Effect of the extracts and fractions of Baccharis trimera and Syzygium cumini on glycaemia of diabetic and non-diabetic mice. Journal of Ethnopharmacology 102, 465469. Orhan, D.D., Aslan, M., Sendogdu, N., Ergun, F., Yesilada, E., 2005. Evaluation of the hypoglycemic effect and antioxidant activity of three Viscum album subspecies (European mistletoe) in streptozotocin-diabetic rats. Journal of Ethnopharmacology 98, 95102. Pari, L., Latha, M., 2005. Antidiabetic effect of Scoparia dulcis: effect on lipid peroxidation in streptozotocin diabetes. General Physiology and Biophysics 24, 1326. Pari, L., Latha, M., 2006. Antihyperlipidemic effect of Scoparia dulcis (sweet broomweed) in streptozotocin diabetic rats. Journal of Medicinal Food 9, 102107. Pascoe, W.S., Storlien, L.H., 1990. Inducement by feeding of basal hyperglycemia in rats with abnormal -cell function. Diabetes 39, 226233. Patel, R., Shervington, A., Pariente, J.A., Martinez-Burgos, M.A., Salido, G.M., Adeghate, E., Singh, J., 2006. Mechanism of exocrine pancreatic insufciency in streptozotocin-induced type 1 diabetes mellitus. Annals of the New York Academy of Sciences 1084, 7188. Pepato, M.T., Mori, D.M., Baviera, A.M., Harami, J.B., Vendramini, R.C., Brunetti, I.L., 2005. Fruit of the jambolan tree (Eugenia jambolana Lam.) and experimental diabetes. Journal of Ethnopharmacology 96, 43 48. Poitout, V., Olson, L.K., Robertson, R.P., 1996. Insulin-secreting cell lines: classication, characteristics and potential applications. Diabetes Metabolism 22, 714. Rajasekaran, S., Sivagnanam, K., Ravi, K., Subramanian, S., 2004. Hypoglycemic effect of Aloe vera gel on streptozotocin-induced diabetes in experimental rats. Journal of Medicinal food 7, 6166. Rajasekaran, S., Sivagnanam, K., Subramanian, S., 2005. Antioxidant effect of Aloe vera gel extract in streptozotocin-induced diabetes in rats. Pharmacological Reports 57, 9096. Rao, N.K., Nammi, S., 2006. Antidiabetic and renoprotective effects of the chloroform extract of Terminalia chebula Retz seeds in streptozotocin-induced diabetic rats. BMC Complementary and Alternative Medicine 6, 17.

T.S. Fr ode, Y.S. Medeiros / Journal of Ethnopharmacology 115 (2008) 173183 Valentovic, M.A., Alejandro, N., Betts Carpenter, A., Brown, P.I., Ramos, K., 2006. Streptozotocin (STZ) diabetes enhances benzo(alpha)pyrene induced renal injury in Sprague Dawley rats. Toxicology Letters 164, 214220. Vuksan, V., Sievenpiper, J.L., 2005. Herbal remedies in the management of diabetes: lessons learned from the study of ginseng. Nutrition, Metabolism, and Cardiovascular Diseases 15, 149160. Wild, S., Roglic, G., Green, A., Sicree, R., King, H., 2004. Global prevalence of diabetes: estimates for 2000 and projections for 2030. Diabetes Care 27, 10471053. Wu, H., Guo, H., Zhao, R., 2006. Effect of Lycium barbarum polysaccharide on the improvement of antioxidant ability and DNA damage in NIDDM rats. Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan 126, 3653671. Wu, K.K., Huan, Y., 2007. Diabetic atherosclerosis mouse models. Atherosclerosis 191, 241249.

183

Yadav, U.C., Moorthy, K., Baquer, N.Z., 2005. Combined treatment of sodium orthovanadate and Momordica charantia fruit extract prevents alterations in lipid prole and lipogenic enzymes in alloxan diabetic rats. Molecular and Cellular Biochemistry 268, 111120. Yu, W.J., Juang, S.W., Chin, W.T., Chi, T.C., Chang, C.J., Cheng, J.T., 2000. Insulin restores neuronal nitric oxide synthase expression in streptozotocininduced diabetic rats. Life Science 68, 625634. Zhao, Y.F., Keating, D.J., Hernandez, M., Feng, D.D., Zhu, Y., Chen, C., 2005a. Long-term inhibition of protein tyrosine kinase impairs electrophysiologic activity and a rapid component of exocytosis in pancreatic -cells. Journal of Molecular Endocrinology 35, 4959. Zhao, R., Li, Q., Xiao, B., 2005b. Effect of Lycium barbarum polysaccharide on the improvement of insulin resistance in NIDDM rats. Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan 125, 981988.