Prunella Vulgaris Ameliorates Liver Injury In Autoimmune Hepatitis Mice And Its Mechanism

Objective:1.To investigate the hepatorenal toxicity in mice with high dose of Prunella vulgaris(200 mg/kg).2.To observe the therapeutic effects of different doses(50,100,200)mg/kg Prunella vulgaris on liver injury in autoimmune hepatitis(AIH)mice.3.To investigate the effects of different doses of Prunella vulgaris on Th1,Th17,regulatory T cells(Treg)in AIH mice.4.To explore the expression of different doses of Prunella vulgaris on the expression of pro-inflammatory factors IFN-γ,IL-17 A and anti-inflammatory factor TGF-β in liver tissues of AIH mice.5.To explore the expression of different doses of Prunella vulgaris on the expression of pro-apoptotic proteins BAX and caspase-3 in liver tissues of AIH mice.Methods:1.Twelve C57BL/6 mice were randomly divided into normal group and high-dose(200mg/kg)Prunella vulgaris group.The normal group were gavaged with 0.2 m L normal saline per day,and the Prunella vulgaris group with 200 mg/kg of Prunella vulgaris instead per day.The diet,activity and mental state of mice were observed daily.30 days later,we detected the levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and urea nitrogen(BUN)by rate method,blood muscle plasma(CRE)by endpoint method,hepatic and renal histopathological changes by HE staining.2.Forty C57BL/6 mice were randomly divided into normal group,AIH model group and 50,100 and 200 mg/kg Prunella vulgaris-treated AIH groups with 8 mice each.AIH mice were established with mouse liver antigen S100 mixed with Freund’s complete adjuvant(S100 /FCA)by intraperitoneal injection.One week after modeling,the normal group and AIH model group were gavaged with 0.2 m L normal saline per day,and Prunella vulgaris-treated AIH groups with 50,100 and 200 mg/kg of Prunella vulgaris instead per day.After 30 days of intervention,we detected the levels of ALT,AST by Rate method,hepatic histopathological changes by HE staining,the protein expression of IFN-γ,IL-17 A and TGF-β by immunohistochemistry,and the protein levels of BAX and caspase-3 by Western blot analysis.Results:1.The high dose Prunella vulgaris had no hepatorenal toxicity in mice: Compared with the normal mice,the activities and the excitability of the mice immediately increased after gavaged the 200mg/kg prunella vulgaris,and the mice returned to normal afterwards,and there was no abnormality in diet and hair.The levels of ALT,AST,BUN,CRE were no difference(p>0.05).The hepatorenal histopathological changes were no obvious changes.2 The therapeutic effect of different doses of Prunella vulgaris on AIH mice:Compared with the normal mice,the liver of AIH mice was color dark,sticky serious,indistinct leaves;the liver lobule structure was disordered.the hepatocyte necrosis was more common,the lymphocyte infiltration was obvious;the levels of ALT,AST were significantly higher(p<0.01).After treatment with various doses of Prunella vulgaris,the liver were crimson,no adhesion,clear leaves;t he hepatic histopathological changes were better than those in the AIH mice,and were dose-dependent;the levels of ALT,AST were decreased in a dose-dependent manner.3.The results of different doses of Prunella vulgaris on Th1,Th17,Treg cells in AIH mice: compared with normal group,the relative percentage of Th1,Th17 cells in spleen of AIH mice were increased significantly(p<0.01),and the relative percentage of Treg cells was decreased significantly(p<0.01);after treatment with various doses of Prunella vulgaris,The relative percentage of Th1,Th17 cells were decreased and the relative percentage of Treg cells was increased.4.The expression of different doses of Prunella vulgaris on IFN-γ,IL-17 and TGF-β in AIH mice: Compared with the normal group,the expression of pro-inflammatory factors IFN-γ and IL-17 A in the AIH model group were increased,and the expression of anti-inflammatory factor TGF-β was decreased.After treatment with various doses of Prunella vulgaris,the expression of pro-inflammatory factors IFN-γ and IL-17 A were decreased,and the expression of anti-inflammatory factor TGF-β was increased,and the color gradient was dose-dependent.5.The expression of different doses of Prunella vulgaris on Bax,caspase3 in AIH mice: Compared with the normal group,the AIH model group had higher expressions of pro-apoptotic proteins Bax and caspase3 in liver tissue.After treatment with various doses of Prunella vulgaris,the expression of Bax and caspase3 were decreased in a dose-dependent manner.Conclusion:Prunella vulgaris can improve liver injury and no obvious hepatorenal toxicity in AIH mice.Its mechanism may be related to the regulation of cell balance Treg/Th17 Prunella vulgaris,anti-inflammatory and anti-hepatocyte apoptosis.The research results provide a new idea for the clinical treatment of AIH with Prunella vulgaris.