To Study The Hepatotoxicity Effect And Mechanism Of Different Processed Products Of Evodia Rutaecarpa (Juss.) Benth

Objective:To compare the hepatotoxicity effect of different processed products of Evodia rutaecarpa（Juss.）Benth,and study the mechanism of Evodia rutaecarpa（Juss.）Benth in different processed ways.Methods:1.According to Chinese Pharmacopoeia（2015）,testing the content of the index component,water and extract of different processed products of Evodia rutaecarpa（Juss.）Benth.2.Taking L02 human normal liver cells as research object,and testing the effect to activities of L02 cells with the method of CCK-8 by processing Evodia rutaecarpa（Juss.）Benth in different ways.Observe the change of cell morphology with inverted microscope and determine the content of alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,lactate dehydrogenase（LDH）,superoxide dismutase（SOD）and malondialdehyde（MDA）in culture supernate.The ratio of SOD/MDA was calculated.Detection the apoptosis with the method of Hoechst fluorescence staining.3.KM mice were divided into control group and the group of different processed products of Evodia rutaecarpa（Juss.）Benth.The raw products,glycyrrhiza processed products and salt processed products of Evodia rutaecarpa（Juss.）Benth were divided into 3 different dose groups.The different dose groups were given in dose of 6.5,3.25,0.65 g·kg^-1 for 5 days,10 days,15 days and then sacrificed.All animals were determined the content of serum alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,lactate dehydrogenase（LDH）and alkaline phosphatase（AKP）.The liver and kidney index were calculated,and the liver histological changes were investigated.15 days animals were tested the content of liver tissue superoxide dismutase（SOD）,malondialdehyde（MDA）,Na⁺-K⁺-ATPase and Ca²⁺-ATPase.Testing the TNF-αand IL-1βlevel of liver tissue with the method of ELISA and determing the Bcl-2、Bax expression of liver tissue with the immunohistochemical.Results:1.The index component test showed that:The content of the index component,water and extract of different processed products of Evodia rutaecarpa（Juss.）Benth accorded with Chinese pharmacopoeia standards.2.The in vitro result showed that:Compared with the control group,the positive group and raw product group of Evodia rutaecarpa（Juss.）Benth can obviously reduce the activities of L02 cells（P<0.01）and morphology,increase the apoptosis of L02 cells.Additionally,the activities of ALT,AST,LDH and MDA were promoted markedly（P<0.05 or P<0.01）,and the activities of SOD were reduced obviously（P<0.05 or P<0.01）.Compared with the raw product group of Evodia rutaecarpa（Juss.）Benth,glycyrrhiza processed group and salt processed group can observably inhibit the proliferation of L02 cells（P<0.01）and the trend on morphological deterioration,decrease the apoptosis of L02 cells.Moreover,the activities of ALT,AST,LDH and MDA were obviously decreased（P<0.05 or P<0.01）,and the activities of SOD were promoted markedly（P<0.05 or P<0.01）.The toxic effects of different processed products of Evodia rutaecarpa（Juss.）Benth on L02 cells in vitro was:the raw product>the glycyrrhiza processed>the salt processed.3.The in vivo result showed that:Compared with the control group,the raw product group of Evodia rutaecarpa（Juss.）Benth can obviously increase the liver and kidney index of mice（P<0.05 or P<0.01）and the damage of liver tissue were increased obviously.Additionally,the activities of serum ALT,AST,LDH and AKP were promoted markedly（P<0.05 or P<0.01）and the activities of liver tissue Na⁺-K⁺-ATPase,Ca²⁺-ATPase and SOD were decreased markedly（P<0.05 or P<0.01）,MDA was increased obviously（P<0.01）.The most importantly,the expression of liver tissue TNF-αand IL-1βwere promoted obviously（P<0.01）,and the percentage of positive cells number of Bcl-2 were significantly decreased（P<0.05 or P<0.01）but Bax were markedly promoted（P<0.05 or P<0.01）.Compared with the raw product group of Evodia rutaecarpa（Juss.）Benth,glycyrrhiza processed group and salt processed group can observably decrease the liver and kidney index（P<0.01,P<0.05）and improved the liver histological.Moreover,the activities of ALT,AST,LDH and AKP were obviously reduced（P<0.01,P<0.05）,and the activities of liver tissue Na⁺-K⁺-ATPase,Ca²⁺-ATPase and SOD were increased obviously（P<0.05 or P<0.01）,MDA was decreased markedly（P<0.01）.Additionally,the expression of liver tissue TNF-αand IL-1βwere decreased obviously（P<0.01）,and the percentage of positive cells number of Bcl-2 were significantly increased（P<0.05 or P<0.01）but Bax were markedly reduced（P<0.05 or P<0.01）.Furthermore,the liver,kidney index and pathologic change showed time dependence.The toxic effects of different processed products of Evodia rutaecarpa（Juss.）Benth on hepatotoxicity in mice was:the raw product>the glycyrrhiza processed>the salt processed.Conclusion:The different processed products of Evodia rutaecarpa（Juss.）Benth were hepatotoxicity in vitro and in vivo,and hepatotoxicity can reduce by processed.The mechanism of Evodia rutaecarpa（Juss.）Benth processing after reducing liver toxicity may relate to reduce free radicals,inflammatory factors,oxidative stress damage and decline in cell apoptosis.