2.Induction of peripheral blood mononuclear cells to hepatocyte-like cells and preliminary study of cell response to injury under the effect of acetaminophen.
Ping LIU ; Biao ZHANG ; Quan ZENG ; Si Wen CHEN ; Chen GE ; Wei Hua WANG ; Chang Zheng WANG ; Wen YUE ; Jun WAN
Chinese Journal of Hepatology 2022;30(1):87-93
Objective: To establish a method for the induction of peripheral blood mononuclear cells to hepatocyte-like cells, and preliminarily investigate cell response to injury under the effect of acetaminophen (APAP). Methods: The surface marker CD45 of peripheral blood mononuclear cells wase detected cells by using flow cytometry and immunofluorescence methods. The cellular morphology of induced hepatocyte-like cells was observed under an inverted microscope. Real-time fluorescent quantitative PCR (RT-PCR) was used to detect the expression level of hepatocyte-specific genes, such as cytochrome (CY) P1A2, CYP3A4, CYP2C9, albumin (ALB), alpha-fetoprotein (AFP), and hepatocyte nuclear factor (HNF)4α mRNA. Immunofluorescence method was used to detect intracellular hepatocyte markers AFP, HNF4α, and ALB expression at the protein level. Biochemical analyzer was used to detect hepatocyte-specific secretory functions of AFP, ALB, and urea. Luciferase chemiluminescence method was used to detect the activity of key drug metabolizing enzyme CYP3A4. Colorimetric assay was used to detect the effect of the drug acetaminophen on hepatocyte-like cells, and alanine aminotransferase (ALT) was used as an indicator of liver cell injury. The statistical differences between the data were compared with t-test and rank-sum test. Results: The positive expression rate of CD45 cell surface markers isolated from peripheral blood mononuclear cells was about 98%, and hepatocyte-like cell morphology changes appeared on 15th day of induction. Compared with isolated mononuclear cells, CYP1A2, CYP3A4, CYP2C9, ALB, AFP and HNF4α mRNA was markedly elevated. The expression level of AFP, ALB and HNF4α protein were equally increased, and the secretory function of AFP, ALB and urea were enhanced. Compared with primary hepatocytes, CYP1A2, CYP2C9, AFP, HNF4α mRNA, and CYP3A4 mRNA did not decrease. The expression levels of AFP, ALB, and HNF4α proteins in the cells did not decrease, and the secretory function of AFP, ALB, and urea did not decrease. In addition, the CYP3A4 enzyme activity produced by hepatocyte-like cells was similar to that of primary hepatocytes. Compared with hepatocyte-like cells incubated without APAP, hepatocyte-like cells incubated with APAP had higher ALT level. Under the effect of APAP, the ALT level of hepatocyte-like cells was higher than isolated mononuclear cells. Conclusion: Peripheral blood mononuclear cells can be induced into hepatocyte-like cells with partial characteristics of hepatocytes, including the activity of CYP3A4, a key enzyme of hepatocyte drug metabolism. Additionally, preliminarily ALT secretory features reflect the hepatocytes injury under the effect of acetaminophen.
Acetaminophen/pharmacology*
;
Cell Differentiation
;
Cells, Cultured
;
Hepatocytes
;
Leukocytes, Mononuclear
;
RNA, Messenger
4.TProtective effect of chlormethiazole, a sedative, against acetaminophen-induced liver injury in mice.
Han Chu LEE ; Sung Ae JUNG ; Hye Kyung JUNG ; Sun Young YI ; Doe Young KIM ; Il Hwan MOON ; Sung Su PARK
The Korean Journal of Internal Medicine 1999;14(2):27-33
OBJECTIVES: The hepatotoxicity of acetaminophen is not a result of the parent compound but is mediated by its reactive metabolite N-acetyl-p-benzoquinone imine. Cytochrome P4502E1 (CYP2E1) is the principal enzyme of this biotransformation, which accounts for approximately 52% of the bioactivation in human microsomes. Recently, chlormethiazole a sedative drug, is reported to be an efficient inhibitor of CYP2E1 activity in human beings. In this study we wished to evaluate whether chlormethiazole, an inhibitor of CYP2E1, could prevent acetaminophen-induced liver injury in mice. METHODS: Acetaminophen, at doses ranging from 200 to 600 mg/kg, was injected into the peritoneum of female C57BL/6 inbred mice fasted for four hours. Chlormethiazole (60 mg/kg) or 5% dextrose water was given 30 min before or 2 h after acetaminophen. Serum aminotransferase activities, histologic index score, survival rate and hepatic malondialdehyde levels were compared. RESULTS: Pretreatment with chlormethiazole 30 min before 400 mg/kg of acetaminophen completely inhibited acetaminophen-induced liver injury (median 118.5 U/L, range 75 to 142 vs. 14,070 U/L, range 5980 to 27,680 for AST; 49 U/L, range 41 to 64 vs. 15,330 U/L, range 13,920 to 15,940 for ALT). In mice receiving chlormethiazole 2 h after acetaminophen, the mean AST and ALT levels were also less elevated, reaching only 20% of the value of acetaminophen-only group. These protective effects were confirmed histologically. Whereas more than 50% of mice died at 500 mg/kg of acetaminophen, all the mice pretreated with chlormethiazole survived at the same dose. CONCLUSION: Chlormethiazole effectively reduces acetaminophen-induced liver injury in mice. Further studies are needed to assess its role in humans.
Acetaminophen/toxicity*
;
Acetaminophen/metabolism
;
Acetaminophen/antagonists & inhibitors
;
Analgesics, Non-Narcotic/toxicity*
;
Analgesics, Non-Narcotic/metabolism
;
Analgesics, Non-Narcotic/antagonists & inhibitors
;
Animal
;
Chlormethiazole/pharmacology*
;
Cytochrome P-450 CYP2E1/antagonists & inhibitors
;
Enzyme Inhibitors/pharmacology
;
Female
;
Human
;
Liver/metabolism
;
Liver/injuries*
;
Liver/drug effects*
;
Mice
;
Mice, Inbred C57BL
;
Sedatives, Nonbarbiturate/pharmacology*
;
Support, Non-U.S. Gov't
5.Experimental study on the pharmacology of 999 ganmaoling, a compound recipe of Chinese and Western materia medica.
Yi-qing MAO ; Zhi-xin MU ; Yue-fei ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(8):726-730
OBJECTIVETo study the pharmacologic characteristics in synergism and complementation of 999 Ganmaoling (GML), a compound recipe composed of Chinese and Western materia medica (CMM & WMM), as well as its theoretical basis of matching of Chinese and Western materia medica.
METHODSThe torsion response induced by glacial acetic acid in mice, toe swelling induced by carrageenanin rats, delayed hypersensitive response in mice and fever induced by endotoxin in rats and rabbits were used to comparatively study the actions of CMM & WMM in GML. The effect of CMM in antagonizing liver damage caused by WD (acetaminophen) in mice was also studied. RT-PCR method was used to analyze the expression of related cytokines.
RESULTSGML showed a significant antipyretic and analgesic effect, it could inhibit the carrageenan induced inflammation, antagonize the endotoxin induced fever, and promote the amount for expression of cytokines in rats' splenic tissue with pneumococci infection to some extent. The CMM in GML showed certain protective effect on acetaminophen induced liver damage.
CONCLUSIONGML has a potent antipyretic, analgesic and anti-inflammatory effects, CMM & WMM in GML showed markedly synergism and complementation, and CMM in it has liver protective effect.
Acetaminophen ; pharmacology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Caffeine ; pharmacology ; Chlorpheniramine ; pharmacology ; Common Cold ; drug therapy ; Drug Combinations ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred ICR ; Rabbits ; Rats ; Rats, Sprague-Dawley
6.A novel class of anti-inflammatory and analgesic drugs--NO-donating NSAIDs.
Yi-hua ZHANG ; Hui JI ; Si-xun PENG
Acta Pharmaceutica Sinica 2007;42(4):352-357
Traditional non-steroidal anti-inflammatory drugs (NSAIDs) and COX-2 selective inhibitors are among the most widely used drugs. However, their significant side effects in gastrointestinal and cardiovascular systems limited the use of these drugs. Recently, research and development of NO-donating NSAIDs (NO-NSAIDs) have become one of the most important strategies to reduce these side effects. NO-NSAIDs may exert a broad range of positive effects in terms of NO-mediated gastrointestinal and cardiovascular safety as well as comparable or increased anti-inflammatory, analgesic properties relative to NSAIDs. This review briefly deals with chemistry of NO-NSAIDs, more details are focused on biological significance, mechanism of action, and therapeutic potential of this novel class of drugs.
Acetaminophen
;
analogs & derivatives
;
chemistry
;
pharmacology
;
Animals
;
Anti-Inflammatory Agents, Non-Steroidal
;
adverse effects
;
pharmacology
;
Aspirin
;
analogs & derivatives
;
chemistry
;
pharmacology
;
Cardiotonic Agents
;
pharmacology
;
Cyclooxygenase Inhibitors
;
adverse effects
;
pharmacology
;
Flurbiprofen
;
analogs & derivatives
;
chemistry
;
pharmacology
;
Gastrointestinal Diseases
;
chemically induced
;
prevention & control
;
Humans
;
Ibuprofen
;
analogs & derivatives
;
chemistry
;
pharmacology
;
Naproxen
;
analogs & derivatives
;
chemistry
;
pharmacology
;
Nitrates
;
chemistry
;
pharmacology
;
Nitric Oxide Donors
;
pharmacology
7.Protective effects of shark hepatic stimulator substance against acute hepatic injury induced by acetaminophen in mice.
Zheng-bing LÜ ; Qian LI ; Bo-ping YE ; Shan BIAN ; Ying WANG ; Qi-ping RUAN ; Wu-tong WU
Acta Pharmaceutica Sinica 2004;39(1):17-21
AIMTo investigate the protective effects of shark hepatic stimulator substance (sHSS) against acute hepatic injury induced by acetaminophen (AAP) in mice.
METHODSAcute hepatic injury model of Balb/c mice was induced by a single intraperitoneal injection of AAP (200 mg.kg-1, i.p.). Serum ALT and AST activities were analyzed. The changes of microstructure and ultrastructure of hepatocyte were observed under optical and electronic microscope. The hepatocyte apoptosis was analyzed by flow cytometer and the expression level of Fas mRNA was determined by RT-PCR.
RESULTSThe activities of serum ALT and AST were significantly decreased and both necrosis and inflammatory infiltration were improved in the mice treated with sHSS 3.0 and 1.5 mg.kg-1. sHSS (3.0 mg.kg-1) prevented the ultrastructural changes of hepatocytes caused by AAP, decreased the percentage of apoptotic cells, and downregulated the expression level of Fas mRNA.
CONCLUSIONsHSS protected hepatocytes from AAP-induced injury, which might be associated with its protection of the mitochondria and inhibition of apoptosis and expression of Fas mRNA in hepatocytes.
Acetaminophen ; Animals ; Apoptosis ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; pathology ; Female ; Growth Substances ; isolation & purification ; pharmacology ; Mice ; Mice, Inbred BALB C ; Peptides ; isolation & purification ; pharmacology ; Protective Agents ; pharmacology ; RNA, Messenger ; genetics ; Random Allocation ; Sharks ; fas Receptor ; biosynthesis ; genetics
8.Hypothetical mode of action of earthworm extract with hepatoprotective and antioxidant properties.
Mariappan BALAMURUGAN ; Kasi PARTHASARATHI ; Lalpet Souri RANGANATHAN ; Edwin L COOPER
Journal of Zhejiang University. Science. B 2008;9(2):141-147
The hepatoprotective potential of earthworm extract (EE) (Lampito mauritii, Kinberg) was evaluated against paracetamol-induced liver injury in Wistar albino rat, in comparison with silymarin, the standard hepatoprotective drug. We observed a reduction in liver antioxidants, such as glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) and in serum total protein, and an increase in serum alkaline phosphatase (ALP), serum aspertate aminotranferase (AST), serum alanine aminotranferase (ALT), bilirubin and liver thiobarbituric acid reactive substances (TBARS) due to liver injury in the paracetamol-administered rats (2 g/kg). On the contrary, increased activities of liver GSH, SOD, GPx, CAT and serum total protein level, and decrease in the contents of serum ALP, AST, ALT, bilirubin and liver TBARS were observed in rats administered with different doses of EE (100, 200 and 300 mg/kg), which are similar to the activities of hepatoprotective drug silymarin (150 mg/kg). The mode of action of EE as evidenced by the above parameters may suggest that EE, on the one hand, prevents the formation of the reactive oxygen groups, or scavenges these groups, thereby preventing the damage on the hepatic cells, and, on the other hand, modulates the genes responsible for synthesis of antioxidant enzymes such as GPx, CAT and SOD in liver tissue and decreases the serum enzymatic activities such as ALP, AST and ALT.
Acetaminophen
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pharmacology
;
Animals
;
Antioxidants
;
metabolism
;
Glutathione
;
metabolism
;
Glutathione Peroxidase
;
metabolism
;
Lipid Peroxidation
;
Liver
;
drug effects
;
metabolism
;
pathology
;
Male
;
Models, Biological
;
Models, Statistical
;
Oligochaeta
;
Oxidative Stress
;
Rats
;
Rats, Wistar
;
Thiobarbituric Acid Reactive Substances
9.Effects of Ziziphus jujuba fruit extracts on cytochrome P450 (CYP1A2) activity in rats.
Xin-Yue JING ; Yun-Ru PENG ; Xin-Min WANG ; Jin-Ao DUAN
Chinese Journal of Natural Medicines (English Ed.) 2015;13(8):588-594
Drug-drug interactions have become a serious problem in the clinic, since plant-based medicines are extensively used. The present study investigated the effects of Ziziphus jujuba fruit (ZJ) extract on the pharmacokinetics of phenacetin, a typical substrate of a cytochrome P450 enzyme CYP 1A2, in rats. The rats were pretreated with the water extract (1.0 g · kg(-1)) or the ethanolic extract (3.6 g · kg(-1)) of ZJ for 10 days, and the pharmacokinetics of phenacetin was investigated after intravenous administration. In an in vitro assay, acetaminophen formation in the hepatic microsomes of ZJ-treated rats was investigated to assess CYP1A2 activity. Our results demonstrated that the treatment with the water and ethanolic extracts of ZJ decreased the plasma concentration of phenacetin and increased the plasma concentration of acetaminophen, resulting in a 43.2% and 15.5% reduction in the AUC0-120 of phenacetin, respectively, and a 53.2% and 64.9% increase in the AUC0-120 of acetaminophen, respectively after intravenous administration. The water or ethanolic extract of ZJ significantly increased the clearance of phenacetin and acetaminophen formation in hepatic microsomes. In conclusion, ZJ extracts displayed effects on the pharmacokinetics of phenacetin and increased the CYP1A2 activity in rats. Therefore, precaution on drug-drug interactions should be taken when ZJ is co-administered with drugs metabolized by CYP1A2, which may result in decreased concentrations of these drugs.
Acetaminophen
;
metabolism
;
Animals
;
Area Under Curve
;
Cytochrome P-450 CYP1A2
;
Cytochromes
;
metabolism
;
Fruit
;
Herb-Drug Interactions
;
Liver
;
drug effects
;
Male
;
Microsomes, Liver
;
Phenacetin
;
metabolism
;
pharmacokinetics
;
Plant Extracts
;
pharmacology
;
Rats, Sprague-Dawley
;
Ziziphus
10.Study on protective effect of water extract from Sabia parviflora on liver injury in mice induced by acetaminophen.
Jian-Qiao LI ; Wen-Feng HUANG ; Hai-Bo HE ; Dong-Xiao ZHAO ; Jian-Zhong HU ; Shun-Guang LU ; Jia-Xin YE ; Kun ZOU
China Journal of Chinese Materia Medica 2020;45(6):1433-1439
The aim of this study was to observe the protective effect of water extract from Sabia parviflora on mice with acute liver injury induced by acetaminophen, and investigate its possible mechanism. Fifty-eight Kunming mice were divided into 6 groups, 8 in the normal group, 10 in the model group, 10 in the biphenyl diester group, and 10 each in the low, medium and high dose groups. After adaptive feeding for one week, the mice in normal group were intragastrically administered with an equal volume of 0.5% sodium carboxymethylcellulose sodium(CMC-Na), and the mice in other groups were intragastrically administered with corresponding drugs at 20 mL·kg~(-1) once a day. Then acetaminophen(200 mg·kg~(-1)) was administered after the above drug administration except the normal group. The behavior and signs of the experimental animals were observed every day and the samples were taken for experiments on the next day of the final administration. The liver mass and mass index were calculated. The blood was collected from the abdominal aorta and centrifuged to obtain the serum for detecting aspartate aminotransferase(AST) activity and alanine aminotransferase(ALT) activity. The liver tissue homogenate was used to detect superoxide dismutase(SOD) activity, glutathione(glutathione, r-glutamyl cysteingl+glycine, GSH) activity and malondialdehyde(MDA) content. Liver tissue was analyzed for histological analysis. The results showed that S. parviflora could alleviate the lipid peroxidation damage in the liver caused by acetaminophen, reduce the ALT and AST activities in serum, increase the levels of SOD and GSH in liver tissue, decrease the content of MDA in liver tissue, and inhibit the apoptosis. S. parviflora could also improve the live histopathological profile, protect liver cells and restore liver function. Among them, the high dose had the most significant effect and showed dose-effect relationship. This study indicated that S. parviflora had a significant protective effect on acetaminophen-induced liver injury in mice, and its mechanism may be related to its anti-oxidation effect and inhi-bitory effect on apoptosis.
Acetaminophen/toxicity*
;
Alanine Transaminase/metabolism*
;
Animals
;
Aspartate Aminotransferases/metabolism*
;
Chemical and Drug Induced Liver Injury/drug therapy*
;
Liver/enzymology*
;
Malondialdehyde/analysis*
;
Mice
;
Oxidative Stress
;
Plant Extracts/pharmacology*
;
Superoxide Dismutase/metabolism*