Compound Glycyrrhizin Tablets Ameliorate Liver Injury Induced by Tripterygium Glycosides Tablet by Regulating Cholesterol Metabolism
Objective:To investigate the mechanism of liver injury induced by tripterygium glycosides tablets(TG)and the molecular mechanism of compound glycyrrhizin tablets(CG)in alleviating the abnormalities of cholesterol metabolism caused by TG via cholesterol metabolism.Methods:According to the body weights,male Sprague-Dawley(SD)rats were randomly grouped as follows:control(pure water),low-dose TG(TG-L,189.0 mg·kg-1·d-1),high-dose TG(TG-H,472.5 mg·kg-1·d-1),TG-L+CG(189.0 mg·kg-1·d-1 TG+20.25 mg·kg1·d-1 CG),and TG-H+CG(472.5 mg·kg1·d-1 TG+20.25 mg·kg1·d-1 CG),with 6 rats in each group.Rats were administrated with corresponding drugs once daily for 3 weeks.At the end of the last administration,the mRNA and protein levels of liver X receptor-alpha(LXR-α),low-density lipoprotein receptor(LDLR),adenosine triphosphate-binding cassette transporter A1(ABCA1),adenosine triphosphate-binding cassette transporter G1(ABCG1),3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGCR),cholesterol 7α-hydroxylase(CYP7A1),cholesterol 12α-hydroxylase(CYP8B1),and sterol 27-hydroxylase(CYP27A1)in the liver tissue were determined by Real-time PCR and Western blotting,respectively.The level of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMG-CoAR),a regulatory enzyme of cholesterol synthesis,was measured by enzyme-linked immunosorbent assay(ELISA).HepG2 cells were used to observe the effect of TG on the cell proliferation in vitro.Specifically,HepG2 cells were grouped as follows:Low-dose TG(TG-1,15 mg·L-1),medium-dose TG(TG-m,45 mg·L-1),high-dose TG(TG-h,135 mg·L1),fenofibrate(FB,10 μmol·L-1),CG extract,TG-h+FB(135 mg·L-1 TG+10 μmol·L-1 FB),TG-m+FB(45 mg·L-1 TG+10 μmol·L-1 FB),TG-l+FB(15 mg·L-1 TG+10 μmol·L-1 FB),TG-h+CG(135 mg·L-1 TG+60 μmol·L-1 CG),TG-m+CG(45 mg·L-1 TG+60 μmol·L-1 CG),and TG-l+CG(15 mg·L-1 TG+60 μmol·L-1 CG).The mRNA and protein levels of LXR-α,ABCG1,LDLR,CYP7A1,CYP8B1,and CYP27A1 in HepG2 cells were determined by Real-time PCR and Western blotting,respectively.Results:The rat experiment showed that compared with the control group,the TG-H group showed down-regulated mRNA levels of CYP7A1,CYP8B1,and CYP27A1 in the liver tissue(P＜0.05,P＜0.01),which were up-regulated by the application of CG(P＜0.05,P＜0.01),and the TG-H+CG group showed up-regulated mRNA level of LDLR(P＜0.01).Compared with the control group,the TG-L and TG-H groups showed down-regulated protein levels of LDLR,CYP7A1,and CYP8B1 in the liver tissue(P＜0.05,P＜0.01).In addition,the protein levels of ABCG1 and LXR-α were down-regulated in the TG-H and TG-L groups,respectively(P＜0.05).Compared with TG alone,TG+CG up-regulated the protein levels of ABCG1 and LDLR(P＜0.05,P＜0.01),and the protein levels of CYP7A1 and CYP8B1 in the TG-H+CG group were up-regulated(P＜0.05,P＜0.01).The cell experiment showed that compared with the control group,the TG-h group presented up-regulated mRNA level of LXR-α(P＜0.01),and the TG-m and TG-h groups showcased down-regulated mRNA levels of LDLR and CYP7A1(P＜0.01)and up-regulated mRNA level of CYP27A1(P＜0.01)in HepG2 cells.The combination of CG with TG restored the above changes(P＜0.01).Western blotting results showed that compared with the control group,the TG-m and TG-h groups showed down-regulated protein levels of LXR-α,ABCG1,LDLR,CYP7A1,CYP8B1,and CYP27A1 in HepG2 cells(P＜0.01).Compared with the TG-h group,the TG-h+CG group showed up-regulated protein level of LDLR(P＜0.05).Compared with the TG-m group,the TG-m+CG group showcased up-regulated protein levels of LDLR,ABCG1,CYP7A1,and CYP27A1(P＜0.05,P＜0.01).Conclusion:The administration of TG at 189.0,472.5 mg·kg-1 for 3 weeks could modulate the signaling pathways associated with cholesterol efflux,endocytosis,and cholesterol biotransformation in hepatocytes,leading to the accumulation of cholesterol and subsequent liver injury in rats.CG could ameliorate the liver injury induced by lipid metabolism disorders caused by TG by up-regulating the expression of LXR-α,LDLR,ABCG1,CYP7A1,CYP8B1,and CYP27A1 to promote cholesterol biotransformation.
tripterygium glycosides tabletscompound glycyrrhizin tabletsliver injurytoxin reduction by compatibilitycholesterol metabolism
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