| Objective The aim of this study is to provide an animal model of the hepatotoxicity of ethanol extracts from Polygonum multiflorum(PMT)in rat liver with similar characteristics shown in humans, based on LPS-induced animal model the influence of Poylgonum multiflorum Thunb on the activity of main subtypes of CYP450 s and the expression of those m RNA and protein during activated in the rats. Then study the infection immune inflammatory signal pathway TLR4 activation and the expression level of downstream signaling molecules My D88, TRIF m RNA and protein to investigate its potential mechanisms.Methods One hundred and thirty male SD rats were randomly divided into eight groups:control, LPS, acetaminophen, LPS+APAP, PMT-L, LPS+PMT-L, PMT-H and LPS+PMT-H group,iv injected weigjt LPS(4mg/kg). Two hours later, according to different groups were orally administered APAP(625mg/kg), PMT-L(6g/kg), PMT-H(12g/kg)respectively, once daily, continuous administration 7day. We observed the change in their weight every day and collected their abdominal aortic blood respectively 2h, 14 h, 5d and 8d for each group.Then detected the biochemical indicators and record the body weight ratio of liver, HE staining histopathological. Furthermore,we detected the activity of CYP1A2, CYP2E1 and CYP3A1 cytochrome in liver cells; reverse transcriptasepolymerase chain reaction(RT-PCR)method was used to analyze the expression of CYP1A2, CYP2E1, CYP3A1, My D88,NF-KB P65, TRIF and IRF-3Mrna. Western-blotting method was used to analyze the expression of CYP1A2, CYP2E1 and CYP3 A, TLR4, My D88, NF-KB P65, TRIF and IRF-3 protein.Results Rats induced by LPS, Compared with the control group, weight loss, liver index increased. First time 2h and 14 h, LPS group, LPS+APAP group and LPS + PMT-L/-H group ALT, AST and ALP increased significantly; histopathological examination revealed that As time went on, Liver injury of the rats in LPS group restored gradually. And on the eight day, the structure of liver tissue of rats in the LPS group was distinct, and the liver cytoplasm appeared slightly red degeneration, LPS + APAP group and LPS + PMT-L/-H focal necrosis of liver cells,with inflammatory cell infiltration, As time went on, LPS group histopathological examination was normal, but LPS+PMT-L/-H group showed significant liver cell degeneration,local chronic inflammatory lesions. Fluorescence assay found in rats induced by LPS in eight days, LPS+PMT-L/-H could significantly reduce rat liver CYP1A2,CYP2E1, CYP3A1 activit; RT-PCR method detected that Polygonum alcohol extract can significantly reduce the large rat hepatic CYP1A2 m RNA expression and significantly increased CYP3A1, My D88, NF-KB P65, TRIF, IRF-3 m RNA expression in rat liver, but no significant effect on CYP2E1 m RNA expression; Western-blotting method detected,Polygonum alcohol extract can significantly reduce rat liver CYP1A2 protein expression,but no significant effect on the expression of CYP2E1 and CYP3A1, significantly increased liver TLR-4, My D88 NF-KB P65, TRIF, IRF-3 protein expression in rat liver.Conclusion It can be well to simulate clinical Polygonum multiflorum Thunb idiosyncratic liver injury induced by LPS, significantly enhanced Polygonum multiflorum Thunb liver ingury in animals model. The occurrence of hepatotoxicity and LPS-induced immune suppression were related to suppression the role of CYP1A2, CYP2E1, CYP3A1 activity and inhibition the expression of CYP1A2 protein and Mrna, and also related to enhance the expression of CYP3A1 m RNA. And that activate TLR4/NF-k B and TLR4/IRF-3 signaling pathway were the mechanisms about the toxicity of PMT by LPS induced in rats liver. |
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