JOURNAL OF MEDICINAL FOOD J Med Food 9 (4) 2006, 587590 Mary Ann Liebert, Inc.

. and Korean Society of Food Science and Nutrition

Short Communication Antihypertensive Effects of Gynura procumbens Extract in Spontaneously Hypertensive Rats
Mi-Ja Kim,1 Hee Jae Lee,2 Sumali Wiryowidagdo,3 and Hye Kyung Kim4
1Department

of Obesity Management, Graduated School of Obesity Science, Dongduk Womens University, Seoul; 2Department of Pharmacology, College of Medicine, Kangwon National University, Chuchon; 4Department of Food and Biotechnology, Hanseo University, Chungnam, Seosan, Republic of Korea; and 3Center for Natural Product Medicine Studies, Department of Pharmacy, University of Indonesia, Depok, Indonesia

ABSTRACT Aqueous extracts of Gynura procumbens (Lour.) Merr. were orally administered to spontaneously hypertensive (SHR) rats for 4 weeks, and antihypertensive effects were determined. Oral administration of 500 mg/kg of G. procumbens (Lour.) Merr. extract (GPE) resulted in significantly lower blood pressure in SHR rats compared with SHR rats not given GPE (P .05). Furthermore, GPE-administered rats had significantly lower serum lactate dehydrogenase, creatine phosphate kinase, and increased nitric oxide (NO), a known vasodilator, compared with the nonGPE-administered SHR group (P .05). These results suggest that oral administration of aqueous GPE may be useful for prevention and treatment of hypertension through increasing NO production in blood vessels. KEY WORDS: creatine phosphate kinase systolic blood pressure

Gymura procumbens extract

lactate dehydrogenase

nitric oxide

INTRODUCTION

including hypertension, arteriosclerosis, heart disease, obesity, diabetes mellitus, and cancers, are increasingly serious public health problems. Circulatory system diseases such as hypertension, arteriosclerosis, and heart disease are especially prevalent in developed countries, with increasing numbers of people showing symptoms of hypertension or prehypertension.1 Hypertension can be caused by many factors, including increases in the volume of body fluid, resistance of the blood vessels, and other factors that elevate blood pressure.2 These factors induce abnormal increases in blood pressure, either alone or in combination. Antihypertensive effects have been noted with various food and natural products.35 Gynura procumbens (Lour.) Merr. is widely distributed in certain areas of Southeast Asia. The leaves of this plant are routinely used in Indonesia for the treatment of kidney diseases, eruptive fevers, rash, hypertension, diabetes mellitus, and hyperlipidemia.6 Zhang and Tan7 suggested that
Manuscript received December 23, 2005. Revision accepted May 25, 2006. Address reprint requests to: Hye Kyung Kim, Department of Food and Biotechnology, Hanseo University, Chungnam, Seosan, 356-706, Republic of Korea, E-mail: hkkim111@hanseo.ac.kr The first two authors contributed equally this study.

IFESTYLE-RELATED DISEASES,

extracts of G. procumbens reduced serum cholesterol and triglyceride levels in streptozotocin-induced diabetic rats. Iskander et al.8 reported that G. procumbens extract (GPE) exhibited anti-inflammatory activities. However, pharmacological mechanisms related to the therapeutic efficacy were not identified. Also, there are no reports in the literature on the hypotensive effect of GPE. Generally, the cytosolic enzymes lactate dehyrogenase (LDH) and creatine phosphate kinase (CPK) as well as lactate in the coronary effluent are important biomarkers for myocardial ischemia in hypertension.9 Therefore, the objective of the present study was to investigate the hypotensive effect of GPE in spontaneously hypertensive (SHR) rats using serum LDH, CPK, and nitric oxide (NO) as biomarkers.

MATERIALS AND METHODS Plant material

G. procumbens (Lour.) Merr. was purchased from KARYASARI (Jakarta, Indonesia). The taxonomy was confirmed by Professor I. Irawati, the chairman of Botanical Department at the Research Center for Biology of the Indonesian Institute of Sciences, Depok, Indonesia. A voucher specimen (number KU-62001) was preserved at the Department of Pharmacology, College of Medicine, Kyung Hee University, Seoul, Republic of Korea.
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Statistical analysis
All data were analyzed and expressed as mean SD. Comparisons were performed by Students t test to detect differences between the groups. A level of P .05 was considered as statistically significant.

RESULTS Effect on SBP

Figure 1 shows the effects of daily oral administration of GPE (500 mg/kg) for 4 weeks on SBP in SHR rats. The GPE group had significantly lower SBP (172.7 14.6 mm Hg) than the control group (191.7 13.2 mm Hg) (P .05).

FIG. 1. Effect of GPE (500 mg/kg) on SBP in SHR rats. Data are expressed as mean SD (bars) values (n 9 rats per group). *P .05 when compared with control.

Effects on LDH and CPK release Preparation of the aqueous extract

The air-dried leaves of G. procumbens were crushed, extracted with 5 volumes of distilled water for 24 hours, and evaporated to dryness in vacuo. The final extract was stored at 20C and diluted with filtered water for each experiment. The effects of GPE on serum activities of LDH and CPK are shown in Figures 2 and 3, respectively. High serum LDH and CPK activities are associated with high blood pressure levels in hypertension.10,11 The oral administration of GPE produced a marked decrease in serum LDH and CPK release by 34% and 48%, respectively, compared with the control group.

Animals
Male SHR rats (10 weeks old) were obtained from Samtako (Osan, Kyungi, Republic of Korea). Animals were fed a commercial rat chow and housed in a controlled environment with a room temperature of 23 1C, relative humidity of 55 10%, and 12-hour light/dark cycle (light from 8:00 to 20:00 h). Body weights of animals at the start of the experiments were 250 12 g. SHR rats were fed an AIN-93-based semipurified standard diet. After preliminary acclimation for 7 days, rats were assigned to two groups of nine rats each and treated as follows: the control group was administered distilled water, and the GPE group was orally administered 500 mg/kg of aqueous GPE one time per day for 4 weeks. Animals were sacrificed by decapitation, and cervical blood was collected. Experiments were conducted in accordance with the animal care guideline of the National Institutes for Health and the Korean Academy of Medical Sciences.

Effect on NO release
The effects of GPE on serum NO concentrations are shown in Figure 4. Concentrations of NO, which acts as a vasodilator, in control and GPE groups were 28.5 3.0 and 45.8 7.2 M, respectively, which indicates that GPE increased the NO concentration by 60.7%.

DISCUSSION
Medicinal herbs have been used as constituents of traditional medicines in Asia for many years.10,12 Most herbs are relatively inexpensive and easily available and have few ad-

Measurement of systolic blood pressure (SBP) and biochemical parameters

The blood pressure was measured noninvasively by a tailcuff method using the Softron BP system (BP-98A, Softron, Tokyo, Japan), after warming the animals at 37C for 10 minutes. At least three determinations were made in every session of SBP measurements, and the mean of three values within 5 mm Hg was taken as the SBP level. Serum LDH, CPK, and NO were determined in serum samples using an autoanalysis system (AU 400, Olympus, Paris, France).

FIG. 2. Effect of GPE (500 mg/kg) on serum LDH in SHR rats. Data are expressed as mean SD (bars) values (n 9 rats per group). *P .05 when compared with control.

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FIG. 3. Effect on oral administration of aqueous GPE (500 mg/kg) on serum lactate CPK in SHR rats. Data are expressed as mean SD (bars) values (n 9 rats per group). *P .05 when compared with control.

verse effects. Accordingly, there is growing interest in the use of herbs and their bioactive compounds. The current study showed a reduction in SBP of SHR following daily oral administration of GPE for 4 weeks, at a dose of 500 mg/kg of body weight. Pressure overload to the heart, such as from hypertension, results in pathological cardiac hypertrophy.13,14 Pathological cardiac hypertrophy induces a reduction of cardiac function,11 and pathological cardiac hypertrophy results in heart failure.14,15 It has been reported that glycolytic energy metabolism is accelerated in hypertensive cardiac hypertrophy induced by pressure overload due to aortic banding or pulmonary hypertension.1619 Furthermore, energy metabolism in hypertensive cardiac hypertrophy in SHR rats, which is a model of hypertension-induced pathological cardiac hypertrophy, shifts away from the use of fatty acid toward the use of glucose.20 The heart is known to rely mostly on fat metabolism for energy, but it can also derive energy from several other substances, including glucose and lactate.21 LDH is found in the cells of almost all body tissues.22 It catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD .23 Cellular injury in tissues containing LDH can result in its release into the bloodstream. Analysis of the different LDH isoenzyme levels in the blood facilitates the diagnosis of some diseases.24 Iemitsu et al.19 reported that mRNA expression of LDH in the glycolytic metabolic pathway in the heart was markedly higher in SHR rats compared with controls. LDH release has been associated with cardiac tissue damage. A higher concentration of LDH could be a symptom of heart damage. We showed that the serum LDH level was significantly lower in GPE (500 mg/kg)-treated SHR rats, suggesting that GPE may protect against cardiac tissue damage. CPK is associated with hypertension. Hropot et al.9 reported that activities of the cytosolic enzymes LDH and CPK significantly increased in rats treated with NO inhibitor, causing a significant increase in SBP. Osbakken et al.25

demonstrated diastolic dysfunction in hypertensive dogs using myocardial CPK enzyme kinetics. An epidemiological study showed that SBP and diastolic blood pressure levels in healthy black and Asian people in the highest serum CPK tertile were 9 and 5 mm Hg higher, respectively, than those in the lowest CPK tertile. In addition, high serum CPK activities and high blood pressure levels were positively correlated in black people.11 The results of the present study demonstrated that GPE significantly decreased serum LDH and CPK release by 34% and 48%, respectively, compared with the control group (P .05), which indicates that GPE may act to reduce blood pressure. Endothelial dysfunction represents an important pathophysiological abnormality that is associated with hypertensive disease.26 One of the important vasodilators released by endothelium is NO. NO is a small ubiquitous molecule that causes vasodilation, and NO concentrations are reduced in the hypertensive state.27 Park et al.28 showed that soy isoflavone supplementation improved SBP in SHR rats by preventing a decrease in NO. Also, Sanchez et al.29 reported that quercetin, a dietary flavonoid, increased endothelial NO synthase, thereby reducing the increase in blood pressure in SHR rats. Hagar et al.30 demonstrated that taurine markedly attenuated renal dysfunction and reduced elevated blood pressure, and increased serum NO levels in cyclosporine Atreated immunosuppressed rats. This is confirmed by findings of impaired NO release in the blood vessels of hypertensive patients, suggesting an important role of NO in the normal control of blood pressure.31 GPE administration caused a significant increase in serum NO concentration in hypertensive rats, indicating it had potent hypotensive effects. In conclusion, our study suggests that aqueous GPE has potential blood pressure-lowering effects in hypertensive rats, which might be mediated by increasing NO production in blood vessels.

FIG. 4. Effect of GPE (500 mg/kg) on serum NO in SHR. Data are expressed as mean SD (bars) values (n 9 rats per group). ***P .001 when compared with control.

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ACKNOWLEDGMENTS
This work was supported by a grant from Kyung Hee University (2002).
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