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Hepatoprotection in a Rat Model of Acute Liver Damage Through Inhibition of CY2E1 Activity by Total
Alkaloids Extracted From Rubus alceifolius Poir
Jiumao Lin, Jinyan Zhao, Tianjiao Li, Jianheng Zhou, Juan Hu and Zhenfeng Hong
International Journal of Toxicology 2011 30: 237 originally published online 11 January 2011
DOI: 10.1177/1091581810390711

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 International Journal of Toxicology
30(2) 237-243
Hepatoprotection in a Rat Model of ª The Author(s) 2011
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Acute Liver Damage Through Inhibition DOI: 10.1177/1091581810390711
http://ijt.sagepub.com
of CY2E1 Activity by Total Alkaloids
Extracted From Rubus alceifolius Poir

Jiumao Lin1, Jinyan Zhao1, Tianjiao Li2, Jianheng Zhou3,

Juan Hu4, and Zhenfeng Hong3

Abstract
We aimed to examine the effect of an alkaloid extract of the roots of Rubus alceifolius Poir on liver damage and cytochrome
enzymes, and underlying mechanism. Hepatotoxicity was induced in rats by treatment with carbon tetrachloride (CCl4). Rats
were then treated with the hepatoprotective drug bifendate, or with low, medium, and high doses of an alkaloid extract from
the roots of R alceifolius Poir. Both bifendate and alkaloid treatment decreased the increase in liver enzymes and cell damage
caused by CCl4. Carbon tetrachloride treatment alone caused a decrease in total cytochrome P450 content, an increase in
CYP2E1 and CYP3A1 messenger RNA (mRNA) levels, and an increase in CYP2E1 and a decrease in CYP3A1 enzymatic
activity. Alkaloid treatment brought these concentrations and activities back toward normal. In summary, these results
suggest that alkaloids from R alceifolius Poir may act to protect the liver through decreasing CYP2E1 enzymatic activity
through decreasing its mRNA.

Keywords
Rubus alceifolius Poir, total alkaloids, CYP2E1, CYP3A1, CYP450, drug metabolic enzyme, acute liver damage

Introduction of action and effect on drug metabolic enzyme concentrations in

Cytochrome P450 enzymes (CYP450) are a superfamily of
mixed function oxidases. In the liver, they are found in the
microsomal fraction and are the major enzymes in phase I
drug metabolism.1 Different isoforms of this enzyme family
have different substrate specificities and different impacts
on metabolism. CYP3A isoforms are the most important class
and are involved in the metabolism of about 50% of drugs.
CYP2E1, another isoform, is particularly important in toxi-
cology because it converts a number of small molecular
weight substances into toxins or carcinogens.2-4 These reac-
tive and toxic intermediates formed during its action increase
the formation of oxygen and hydroxyl free radicals5 that dam-
age cellular structure and cause hepatotoxicity.6,7 In addition,
liver damage has been shown in some studies to alter the
activity of CYP450 enzymes.8
Rubus alceifolius Poir belongs to the Rubus genus of the
Rosaceae plant family. Its roots and leaves have a plain flavor
and sweet taste, and it has been used as an herbal medicine in
applications such as diuresis, detoxification, and bruise healing.
It is also widely used to treat various types of hepatitis by the
local population in Anxi, Southern Fujian. Previous studies by
the present research group have shown R alceifolius Poir to have
very good hepatoprotective effects.9-12 However, its mechanism
the liver is unknown. The study reported here aims to investigate
the effect of total alkaloids extracted from R alceifolius Poir on
both the messenger RNA (mRNA) expression and the enzyme
activity of CYP2E1 and CYP3A1, and thus to provide insight
into the mechanism for the hepatoprotective effects of these
alkaloids. In this study, liver damage was produced with car-
bon tetrachloride (CCl4), and the effects of bifendate (a clini-
cally used hepatoprotective drug) and low-, medium-, and
high-dose alkaloid treatment on biological markers of liver
damage, on total CYP450 enzymes, and on CYP2E1 and
CYP3A1 mRNA and enzyme activity were compared.
1
Fujian Academy of Integrative Medicine, Fujian University of Traditional
Chinese Medicine, Fuzhou, Fujian, China
2
The Second Affiliated Hospital of Fujian University of Traditional Chinese
Medicine, Fuzhou, Fujian, China
3
Department of Integrative Medicine of Fujian University of Traditional
Chinese Medicine, Fuzhou, Fujian, China
4
Department of Pharmacy of Fujian University of Traditional Chinese
Medicine, Fuzhou, Fujian, China
Corresponding Author:
Zhenfeng Hong, Department of Integrative Medicine of Fujian University of
Traditional Chinese Medicine, Fuzhou, Fujian 350108, China
Email: zfhong1953@163.com

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238
Materials and Methods
Animals
In all, 60 Sprague Dawley rats 180 to 220 g (30 males and
30 females) were provided by Shanghai Experimental Animal
Co Ltd, China, and allowed to acclimate in our animal facility
for 5 days before use. The study was approved by the institu-
tional review board of the Laboratory Animal Center of Fujian
University of Traditional Chinese Medicine. All experiments
were carried out under the guidelines of the Laboratory Proto-
col of Animal Handling of our college.
Preparation of Alkaloids and Assay of
Total Alkaloid Content
The roots of R alceifolius Poir were collected from Anxi of
Fujian Province, identified and authenticated by experts in our
College, and extracted the alkaloids as follows.13 The herb pow-
der (1 g) was extracted with 50 mL chloroform:methanol:ammo-
nia solution (15:4:3) for 2 hours in an ice bath, sonicated for
30 min, brought to room temperature and filtered. The filteed
solution was collected and dessicated. The resultant residue was
dissolved by 2 mL of 2% sulfuric acid solution and filtered. The
filter paper and residue were re-washed with 2 mL of 2% sulfuric
acid solution and with buffer solution (pH 3.6). Buffer was then
added to make a final volume of 50 mL, and the solution saved
for future use. Acid dye colorimetry was used to measure total
alkaloid content. Total alkaloid content was 0.81 mg of alkaloid
per gram of initial herb powder.
Reagents and Equipment
Reagents. Carbon tetrachloride was obtained from
Chinese United Chemical Reagent Co, Ltd (Shanghai, China),
erythromycin (a CYT3A1 substrate) from Shenggong Bio-
Engineering Technology Co (Shanghai, China), aminopyrazo-
lone morpholine (a CYT2E1 substrate) from Shanghai Reagent
Factory (Shanghai, China), NADPH from Roche (Basel,
Switzerland); and Trizol from Invitrogen (Carlsbad, California).
Bifendate (dimethyl diphenyl bicarbonate) pills were purchased
from IMC Pharmaceutical Co, Ltd (Zhejiang, China) and diluted
to 1% concentration for application.
Alanine aminotransferase/Glutamic-pyruvic transaminase
(ALT/GPT) and aspartate aminotransferase/Glutamic-oxaloacetic
transaminase (AST/GOT) detection kits were obtained from Jian-
cheng Bio-engineering Institute (Nanjing, China), Bradford pro-
tein quantification and detection kit from Shenggong Bio-
Engineering Technology Co, Ltd (Shanghai, China), deoxyribo-
nucleoside triphosphate (dNTP) and reverse transcription (RT) Kit
from MBI Fermentas Inc (Glen Burnie, Maryland), and Taq
polymerase and RNAse inhibitor from TaKaRa Biotechnology
(Otsu, Shiga, Japan). Primers were synthesized by Shanghai Hand-
some Bio Co, Ltd (Shanghai, China). Terminal deoxynucleotidyl
transferase dUTP nick end labeling (TUNEL) staining kits and
related reagents were purchased from Maixin Ltd (Fujian, China).
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International Journal of Toxicology 30(2)
Equipment. The following instruments were used: Du650
Spectrophotometer and 64R High-Speed Refrigerated Bench-
top Centrifuge (Beckman, USA), BT 224 semiautomated spec-
trophotometer (Biotechnica, Italy), PCR-9600 (Perkon-Elmer,
USA);GEL DOC 2000 gel imaging system (Bio-Rad), Sanheng
multipurpose electrophoresis instrument (Beijing 61 Instru-
ment Factory, China).
Animal Protocol, Drug Treatment, and Tissue Preparation
Animal protocol. In all, 60 rats were randomly divided into the
following groups (5 males and 5 females in each group): saline,
liver damage, and bifendate groups, and low-, medium-, and
high-dose alkaloid treatment groups. Rats were given intraperi-
toneal (ip) injections of 0.2 mL saline (saline group), or 0.1 mL/
100 g body weight of 99.5% CCl4 ip (liver damage, bifendate,
and alkaloid groups) in order to induce liver damage. After a
24-hour period to allow the biochemical changes seen in CCl4-
induced liver damage to appear,8 rats were given intragastric
administration of saline (saline and liver damage groups),
0.1g/kg body weight of 1% bifendate (bifendate group),9,10 or
alkaloid in a dose of 0.36 g/kg per d, 0.72 g/kg per d, or 1.44
g/kg per d (low-, medium-, and high-dose alkaloid groups,
respectively). These doses were given once daily for 7 days.
Tissue and microsome preparation. After the last drug or saline
treatment, rats were fasted (but given water) for 24 hours and
then sacrificed. For measurement of indicators, 5 mL blood
samples were collected, centrifuged, and the serum removed
and stored at 20 C. The liver was removed, cleansed with
cold saline, divided into 2 pieces, and stored at 80 C.
A small piece of tissue from the central part of the right lobe
(about 0.5 cm3) was fixed in 10% formalin immediately for
routine sectioning, hematoxylin and eosin staining, and histo-
pathological examination.
Liver microsomes were prepared by differential centri-
fugation, suspended in Tris-HCl buffer (1 mL/g), and stored
at 70 C until further use.
Measurement Methods
Determination of transaminase activity. Serum AST and ALT
activities were measured with a BT 224 semiautomated spec-
trophotometer according to the instruction kit.
Histopathological examination of liver tissue. Morphological
change in liver tissue was examined under a light microscope.
Pathological changes in liver tissue were divided into 5 grades,
as follows (Figure 1): normal cells, ‘‘’’; spotty necrosis, ‘‘þ’’;
focal necrosis, ‘‘þþ’’; necrosis in <one third of the cells in a
lobule, ‘‘þþþ’’; and necrosis in >one third of the cells in a
lobule, ‘‘þþþþ.’’
Measurement of microsomal protein concentration. Microsomal
protein concentrations were measured with the Bradford Pro-
tein Assay Kit, according to the manufacturer’s instructions.
Lin et al 239

Figure 1. Grading system used to assess hepatic histopathological damage. (A) ‘‘’’ normal liver cells; (B) ‘‘þ’’ spotty necrosis; (C) ‘‘þþ’’ focal
necrosis; (D) ‘‘þþþ’’ necrosis in less than one third of the cells in a lobule; (E) ‘‘þþþþ’’ necrosis in more than one third of the cells in a lobule.

Quantification of total CYP450 enzyme levels. The concentra-
tion of total CYP450 enzymes was measured with a modified
method of Zheng et al.14 To 2 mL of buffer, 100 mL of
microsome suspension and 0.5 mL of 10% sodium dithionite
solution were added, mixed immediately, loaded into a colori-
metric cup, and filled with CO for 30 seconds. A second 100 mL
aliquot of the suspension was added to 2 mL of buffer, imme-
diately mixed, and loaded into another colorimetric cup to
serve as control. Optical density (OD) values at 450 nm and
490 nm wavelengths were recorded; and the CYP content cal-
culated according to the following formula:
Total CYP450 content ðnmol=mgÞ ¼ D OD ð450 nm  490 nmÞ
 10:99=protein concentration:
Measurement of CYP2E1 and CYP3A1 enzymatic activity. Ami-
nopyrazolone morpholine-N-demethylase activity was used to
measure CYP2E1 activity,15 and erythromycin demethylase
activity was used to measure CYP3A1 activity. To measure
CYP2E1 activity, 20 mg/mL aminopyrazolone morpholine-
N-demethylase was added to microsomes suspended in
0.02 mol/L Hepes buffer and the solution was kept at 37 C
in a water bath for 3 minutes; 0.01 mL of 0.1 M NADPH was
then added and the solution was kept in 37 C water bath for
Table 1. Primers for CYP3A1, CYP2E1, and b-Actin
Product
Name Primers (bp)
CYP3A1 Sense: 50 -ATCCGATATGGAGATCAC-30 579
Antisense: 50 -GAAGAAGTCCTTGTCTGC-30
CYP2E1 Sense: 50 -CTCCTCGTCATATCCATCTC-30 473
Antisense: 50 -GCAGCCAATGAGAATGTGG-30
b-actin Sense: 50 -GTTCGCCATGGATGACGATATC-30 265
Antisense: 50 -GCCAGATCTTCTCCATGTCGT-30
30 minutes. The next steps were the same as those performed
in making a formaldehyde standard curve. To measure erythro-
mycin demethylase activity, the same procedure was used
except for replacing aminopyrazolone morpholine-N-demethy-
lase with 0.004 mol/L erythromycin demethylase. Both activi-
ties were calculated as formaldehyde production compared to a
formaldehyde standard curve.
Expression of target gene mRNA. The expression of
CYP3A1mRNA and CYP2E1mRNA was measured by reverse
transcription ploymerase chain reaction (RT-PCR). The ampli-
fied product was separated by 1.5% agarose gel electrophor-
esis; and the band analyzed with a gel image processing

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240 International Journal of Toxicology 30(2)

Table 2. Summaries of Weights and Liver Function for the Rats in the 6 Groups
Groups

Control (n ¼ 10) CCl4 (n ¼ 10) Bifendate (n ¼ 10) TA 3.6% (n ¼ 10) TA 7.2% (n ¼ 10) TA 14.4% (n ¼ 10) P Value

Weight (g) 201.0 (198.0, 205.0) 201.0 (197.0, 202.0) 197.0 (188.0, 208.0) 201.5 (193.0, 206.0) 205.5 (192.0, 207.0) 201.5 (199.0, 206.0) .947
ALT (U/L) 22.2 (11.8, 22.9) 90.6 (83.1, 104.3)a 58.9 (45.7, 64.5)a,b 60.3 (45.4, 69.2)a,b 55.8 (46.4, 66.6)a,b 45.1 (42.2, 46.4)a,b,e <.001f
AST (U/L) 60.3 (50.3, 68.4) 102.1 (95.2, 111.1)a 78.8 (75.2, 88.8)a,b 89.3 (79.4, 101.2)a 59.2 (57.6, 73.7)b,c,d 61.1 (53.6, 76.3)b,c,d <.001f
Degree of pathology liver injury
 10 (100.0%) 0 (0.0%) 1 (10.0%) 0 (0.0%) 0 (0.0%) 1 (10.0%) <.001f
þ 0 (0.0%) 0 (0.0%) 4 (40.0%) 3 (30.0%) 3 (30.0%) 5 (50.0%)
þþ 0 (0.0%) 0 (0.0%) 3 (30.0%) 5 (50.0%) 6 (60.0%) 3 (30.0%)
þþþ 0 (0.0%) 1 (10.0%) 2 (20.0%) 2 (20.0%) 1 (10.0%) 1 (10.0%)
þþþþ 0 (0.0%) 9 (90.0%) 0 (0.0%) 0 (0.0%) 0 (0.0%) 0 (0.0%)

Abbreviations: ALT, alanine aminotransferase; AST, aspartate aminotransferase; TA, total alkaloid; , normal cells; þ, spotty necrosis shown in a few cells; þþ,
focal necrosis shown in a small area; þþþ, necrosis area shown in < one third of the cells in a lobule; þþþþ, necrosis area shown in > one third of the cells in a
lobule
a
Indicates a significant difference compared to the control group.
b
Indicates a significant difference compared to the CCl4 group.
c
Indicates a significant difference compared to the bifendate group.
d
Indicates a significant difference compared to the TA 3.6% group.
e
Indicates a significant difference to the compared to the TA 7.2% group.
f
P value <.05 indicates a significant difference among the 6 groups.

system. b-actin was used as a housekeeping gene. The primers
used are shown in Table 1.
Apoptosis index. The apoptosis index (AI) was determined by
TUNEL staining (in situ end labeling, performed with the
TUNEL detection kit, according to the kit instructions). The
proportion of apoptotic cells in each group was recorded to
determine the AI. After staining, 5 high-power fields (400)
were randomly selected in each slide and the average propor-
tion of positive cells in each field counted and classified into
5 grades: grade 0: <2%, grade 1: <3% to 9%, grade 2: <10%
to 19%, grade 3: <20% to 39%, and grade 4: >40%.16
Statistical Analysis
For continuous variables, data are shown as median and inter-
quartile range (IQR). The comparisons between the 6 groups
were performed using the Kruskal-Wallis test, and the compar-
isons between 2 groups by the Mann-Whitney U test. For cate-
gorical variables, data are presented by number and percentage.
The associations between categorical variables were tested
using Fisher exact test. All statistical hypothesis tests were set
with a significance level of .05. Statistical analyses were per-
formed using Statistical Package for the Social Sciences
(SPSS) 15.0 statistics software (SPSS Inc, Chicago, Illinois).
Results
Effect of CCl4, Bifendate, and Alkaloids on
Biochemical and Histological Liver Damage
Table 2 shows weights, liver enzymes, and degree of liver
pathology, and Figure 2A shows the AI, in the 6 groups of rats.
Liver enzymes. Serum ALT and AST levels in the liver injury
group (CCl4 treatment alone) were significantly elevated over
those of the control group (saline treatment). When bifendate was
given after CCl4 treatment, ALT and AST levels were signifi-
cantly reduced compared to those in the liver injury group, but
remained significantly higher than those of the control group.
Alkaloid administration caused a dose-dependent decrease in
both ALT and AST levels from those seen in the liver injury
group. Aspartate aminotransferase levels were decreased to con-
trol levels by both medium- and high-dose alkaloid. However,
ALT levels, although decreased from the levels in the liver
damage group, were still twice those of the control group at the
highest alkaloid dose used.
Liver histology. Histological examination showed CCl4 treat-
ment to produce severe acute liver damage (Table 2). While
no control rats showed histological signs of liver damage, 9 of
the 10 rats in the CCl4-treated, liver damage group were in the
highest damage category (> one third of the cells necrotic). No
bifendate or alkaloid treated rat had liver damage of this severity.
Both types of treatment lessened the liver damage caused by
CCl4, and the effect of alkaloid treatment was dose-dependent.
Apoptosis. The TUNEL assay records DNA fragmentation, an
indication of apoptosis and cell death. As expected, CCl4 treat-
ment produced a significant increase in the AI obtained from this
assay. Bifendate and alkaloid treatment both decreased the
degree of DNA fragmentation caused by CCl4, but not to control
levels. (Figure 2A)
Effect of CCl4, Bifendate, and Alkaloid on CYP450,
CYP2E1, and CYP3A1
The results described above show CCl4 treatment to cause sub-
stantial acute liver damage and both bifendate and alkaloid
treatment to alleviate this damage, although not completely.
Figures 2 and 3 show the effects of CCl4, bifendate, and alka-
loid on liver cytochromes.

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Lin et al 241

Figure 2. Box plots for apoptosis index (TUNEL assay), CYP450 concentration, CYP2E1 and CYP3A1 enzymatic activity, and mRNA
expression in the 6 experimental groups. (A) apoptosis index (TUNEL assay); (B) CYP450 concentration; (C) CYP2E1 activity; (D) CYP2E1
mRNA expression; (E) CYP3A1 activity; (F) CYP3A1 mRNA expression. TA indicates total alkaloid; TUNEL, terminal deoxynucleotidyl
transferase dUTP nick end labeling; a, significant difference compared to control group; b, significant difference compared to CCl4 group; c,
significant difference compared to bifendate group; d, significant difference compared to TA 3.6% group; e, significant difference compared
to TA 7.2% group.

CYP450 concentration. Carbon tetrachloride treatment although at the low dose did not change CYP450 levels over
decreased total CYP450 enzyme levels (Figure 2B). Bifendate those seen with CCl4-treatment alone, at the high dose raised
increased the levels of this enzyme in CCl4-treated animals, these levels completely back to those seen in normal, saline-
but not completely back to control levels. Alkaloid treatment, treated rats.

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242
Figure 3. Expression of CYP2E1 and CYP3A1 mRNA, shown by
electrophoresis, is increased in liver damage and decreased by alkaloid
treatment. A indicates control group; B, liver damage group;
C, biphendate group; D, alkaloid low-dose group; E, alkaloid
medium-dose group; F, alkaloid high-dose group
CYP2E1 enzymatic activity and mRNA levels. Although CCl4
treatment decreased total CYP450 protein levels, this treatment
increased the enzymatic activity of CYP2E1 (Figure 2C).
Bifendate had no effect on this increase. Alkaloids, in contrast,
reversed the increase in a dose-dependent manner to such an
extent that CYP2E1 enzymatic activity was not only reduced
to levels lower than those of the liver injury group, but at the
highest dose, to levels significantly lower than those seen in
the control group. CYP2E1 mRNA expression showed a pat-
tern similar to that seen in enzymatic activity (Figure 2C and
D, Figure 3). Carbon tetrachloride treatment significantly
increased mRNA; bifendate had no effect; and alkaloid treat-
ment caused a dose-dependent decrease to levels, at the high-
est dose, that were similar to control (Figures 2C, Figure 3).
CYP3A1 enzymatic activity and mRNA levels. Although
CYP2E1 enzymatic activity and mRNA levels changed in a sim-
ilar direction during CCl4, bifendate, and alkaloid treatment,
CYP3A1 enzymatic activity and mRNA levels changed in oppo-
site directions (Figure 2D and E, Figure 3). Carbon tetrachloride
treatment significantly increased CYP3A1 mRNA levels, but
significantly decreased its enzymatic activity. Alkaloid treat-
ment completely reversed the changes made by CCl4, that is,
alkaloid increased enzymatic activity and decreased mRNA, and
in doing so brought both parameters back to normal levels.
Bifendate caused a similar, but smaller, reversal of CCl4-induced
changes in enzymatic activity and mRNA.
Discussion
We have shown in this study that an extract of alkaloids from
R alceifolius Poir blocks the increase in CYP2E1 mRNA and
enzyme activity that occurs in liver injury caused by CCl4, a clas-
sical model used for drug screening and the study of hepatitis.
This action is likely to be the major mechanism by which these
alkaloids produce their known hepatoprotective effects.
Reactive intermediates formed during the CYP2E1-induced
metabolism of CCl4 are responsible for the increase in reactive
oxygen species (ROS) that causes the liver damage. In the
present study, we found, as have others,4,17,18 that although
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International Journal of Toxicology 30(2)
total CYP450 protein was decreased during CCl4-induced liver
damage, CYP2E1 activity was increased. Decreased antioxi-
dant capacity also occurs in CCl4-induced liver injury. Thus,
both an increased ability to generate ROS and a decreased abil-
ity to detoxify these species are involved.
Drug-induced liver injury is usually caused by excessive
formation of ROS. These ROS cause apoptosis, an inflamma-
tory response, and oxidative stress. Oxidative stress, among
other actions, increases lipid peroxidation.19,20 Our previous
studies have shown that extracts of R alceifolius Poir effec-
tively inhibit the formation of ROS, inflammatory cytokines,
and lipid peroxidation.9-12,21
We have also reported previously that alkaloid extracts of this
plant decrease both histological and biochemical signs of hepatic
injury,22 and more recently have reported that alkaloid extracts
decrease the mRNA of 2 P450 isoforms, CYP 2E1 and CYP
3A1.23 In the current study, we wished to examine the enzyme
activity of CYP2E1 and CYP3A1 in addition to mRNA level and
the evidence of alkaloid protective action against liver damage,
and thus provide a comprehensive picture of Rubus alkaloid
actions and the underlying mechanism responsible for them.
In searching for a causative mechanism, we found Rubus
alkaloids to prevent both the increases in CYP2E1 mRNA
and the increases in CYP2E1 activity seen in CCl4-induced
liver damage, thus providing a likely explanation for their
hepatoprotective action. An effect of Rubus alkaloids on
CYP2E1 activity has not been previously reported. A recent
report on treatment of rats with iridoid glucosides from
Boschniakia rossica, an antioxidant and anti-inflammatory
herbal medicine, stated that CCl4 treatment decreased
CYP2E1 protein and activity and iridoid glucosides increased
both.24 And, negundeside, another herbal extract, has been
reported to have no effect on CYP2E1, but instead, to
increase antioxidant activity, and thus to act by an alternate
mechanism for hepatoprotection.25
Our data suggest that the Rubus alkaloids are hepatoprotec-
tive through their action in decreasing CYP2E1 mRNA and
therefore CYP2E1 activity. But the actions of CCl4 and of these
alkaloids on the other isoform studied, CYP3A1, are a puzzle,
for CCl4 increased CYP3A1 mRNA but decreased its activity,
and Rubus alkaloids decreased CYP3A1 mRNA and increased
its activity. One possible explanation is that some toxic product
from the increased CYP2E1 metabolic activity or perhaps from an
initially increased CYP3A1 activity is an inhibitor of CYP3A1.
This difference between the effect of CCl4 on CYP2E1 and
CYP3A2 should be investigated further, keeping in mind that
compounds that change specific CYP isoform activity in rats may
not do so in humans, or may do so by different mechanisms.
In interpreting our results, one must remember that a change
in the synthesis of cytochrome mRNA is not the only way that
liver damage due to ROS is controlled. Stabilization of the exist-
ing mRNA can play a role. Alteration in 1 of the 2 pathways
for cytochrome degradation (the slow lysosomal pathway or the
faster proteosomal pathway) may also change the level of cyto-
chrome activity.26 In addition, changes in the levels of antioxidant
substances are also involved in control of damage from ROS.
Lin et al
The results of the present study may be meaningful to guide
proper drug treatment and to understand drug interactions and
their metabolism inside the body. It helps to clarify the mole-
cular mechanisms and targets for traditional Chinese medicine
and provides new ideas and methods for the modernization of
Chinese medicine study. We have recently performed a study
to isolate and identify the active constituents of R alceifolius
Poir extracts and found the constituent 1b-hydroxyeuscaphic
acid to have the strongest effect against CCl4-induced hepato-
toxicity.27 But no investigation as to the mechanism has yet
been performed. And drug metabolism and processing of bio-
logical effects in the body is a very complex subject and
requires much further study.
Declaration of Conflicting Interests
The author(s) declared no potential conflicts of interests with respect
to the authorship and/or publication of this article.
Funding
The author(s) disclosed receipt of the following financial support for
the research and/or authorship of this article: The project was sup-
ported by the Nature Science Foundation of Fujian Province of China
(No.2006J0109); The Project Sponsored by Open Fund of Fujian Key
Laboratory of Integrative Medicine on Geriatrics (Fujian University of
TCM)(No. 2008J1004-19 and 2008J1004-17); Developmental Fund
of Chen Keji Integrative Medicine (CKJ2008054 and CKJ2008056).
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