Protective Effect Of Gardenia Jasminoides Ellis Aqueous Extract On Acute Chemical Injury And Drug-induced Liver Injury In Mice

Objective:To investigate the protective effect and molecular mechanism of Gardenia jasminoides Ellis aqueous extract（SZZJYA）on chemical injury and drug-induced liver injury in mice.Methods:（1）Protective effect of SZZJYA in mice acute CCl₄-Liver injury:The model of acute liver injury was established by intraperitoneal injection of CCl₄.The Kunming mice was stochastically divided into six groups:normal control group（NC）,model group（MC）,high-,medium-and low-doses of SZZJYA group,and positive control group（Biphenyl,DDB）.NC and MC groups were treated with peanut oil,the positive control group and all doses of SZZJYA groups were treated with DDB（150mg/kg）and SZZJYA（5.00,2.50,1.25g/kg）respectively.All the mices were intragastrically administered with drugs or peanut oil once a day for 7 days.NC group was injected with peanut oil,acute chemical injury in the other groups were induced by intraperitoneal injected 0.18%CCl₄（0.18%CCl₄,0.018ml/kg）in peanut oil 1 hour after the last administration.Then all the mices were weighed,took the blood and draw the materials after 20h.Then the coefficient of the liver was determined.The activation of ALT,AST,ALP,the concentration of TBIL,TP,ALB,GLB,A/G in serum,the activation of SOD,GSH-PX and the concentration of MDA,GSH in liver tissue were measured by spectrophotometry.HE stain was used to determine pathological changes of hepatic injury.The immunohistochemical method was used to check the protein expressions of PNCA in liver tissue.（2）Protective effect of SZZJYA in mice acute D-Gal N-Liver injury:The model of acute liver injury was established by intraperitoneal injection of D-GalN.The Kunming mice was stochastically divided into six groups:NC group,MC group,SZZJYA_H group,SZZJYA_M group,SZZJYA_L group and Biphenyl group.NC and MC groups were treated with NS,Biphenyl group and all doses of SZZJYA groups were treated with the corresponding drug respectively.All the mices were intragastrically administered with drugs or NS once a day for 7 days.NC group was injected with NS,acute chemical hepatic injury in the other groups were induced by intraperitoneal injected in D-GalN（700mg/kg）1 hour after the last administration.Then all the mices were weighed,took the blood and draw the materials after 20h.The coefficient of the liver was determined.The activation of ALT,AST,ALP,the concentration of TBIL,TP,ALB,GLB,A/G in serum,the activation of SOD,GSH-PX and the concentration of MDA,GSH in liver tissue were measured by spectrophotometry.HE stain was used to determine pathological changes of hepatic injury.The immunohistochemical method was used to check the protein expressions of Bax and Bcl-2 in liver tissue.（3）Protective effect of SZZJYA in mice acute APAP-Liver injury:The model of acute liver injury was established by intraperitoneal injection of APAP.The Kunming mice was stochastically divided into six groups:NC group,MC group,SZZJYA_H group,SZZJYA_M group,SZZJYA_L group and Biphenyl group.NC and MC groups were treated with NS,Biphenyl group and all doses of SZZJYA groups were treated with the corresponding drug respectively.All the mices were intragastrically administered with drugs or NS once a day for 7 days.NC group was injected with NS,acute chemical hepatic injury in the other groups were induced by intraperitoneal injected in APAP（120mg/kg）1 hour after the last administration.Then all the mices were weighed,took the blood and draw the materials after 20h.The coefficient of the liver was determined.The activation of ALT,AST,ALP,the concentration of TBIL,TP,ALB,GLB,A/G in serum,the activation of SOD,GSH-PX and the concentration of MDA,GSH in liver tissue were measured by spectrophotometry.HE stain was used to determine pathological changes of hepatic injury.The immunohistochemical method was used to check the protein expressions of TNF-αand iNOS in liver tissue.Results:（1）Protective effect of SZZJYA on acute CCl₄-Liver injury in mice:Compared with model group,different doses of SZZJYA groups could increase weight and decrease the coefficient of the liver,spleen and thymus（P<0.01 or P<0.05）,SZZJYA_M,SZZJYA_H groups could significantly decreased the coefficient of the liver,spleen and thymus（P<0.01）,The activation of ALT,AST,ALP and the concentration of TBIL in serum were reduced.The concentration of TP,ALB,GLB and the ratio of A/G in serum were raised（P<0.01 or P<0.05）,SZZJYA_M,SZZJYA_H groups could significantly decreased the activation of ALT and AST（P<0.01）,Different doses of SZZJYA groups could reduce the concentration of MDA in hepatic homogenate and raise the activation of SOD,GSH-PX,the concentration of GSH in liver tissue（P<0.01 or P<0.05）,SZZJYA_M,SZZJYA_H groups could have the most of protective effect（P<0.01）,Different doses of SZZJYA groups could alleviate liver injury（P<0.01 or P<0.05）,Immunohistochemical result showed that different doses of SZZJYA groups could raise the expression of PCNA protein（P<0.01）.（2）Protective effect of SZZJYA on acute D-GalN-Liver injury in mice:Compared with model group,different doses of SZZJYA groups and could increase weight and decrease the coefficient of the liver,spleen and thymus significantly（P<0.01）,The activation of ALT,AST,ALP and the concentration of TBIL in serum were reduced.The concentration of TP,ALB,GLB and the ratio of A/G in serum were raised（P<0.01 or P<0.05）.SZZJYA_M,SZZJYA_H groups could significantly decrease the activation of ALP and the concentration of TBIL,and raise the ratio of A/G（P<0.01）,The concentration of MDA in hepatic homogenate was reduced,and the activation of GSH-PX and the concentration of GSH were raised in liver tissue by SZZJYA_M,SZZJYA_H groups treatment（P<0.01）,Different doses of SZZJYA groups could alleviate liver injury（P<0.01 or P<0.05）,Immunohistochemical result showed that different doses of SZZJYA groups could increase the expression of Bax and Bcl-2 protein（P<0.01）.（3）Protective effect of SZZJYA on acute APAP-Liver injury in mice:Compared with model group,different doses of SZZJYA groups could increase weight and decrease the coefficient of the liver,spleen and thymus（P<0.01or P<0.05）.The weight of the mice and the coefficient of the liver,thymus could be affected by SZZJYA_M,SZZJYA_H groups treatment significantly（P<0.01）,The activation of ALT,AST,ALP and the concentration of TBIL in serum were reduced.The concentration of TP,ALB,GLB and the ratio of A/G in serum were raised（P<0.01 or P<0.05）.SZZJYA_M,SZZJYA_H groups could have the most of protective effect（P<0.01）,Different doses of SZZJYA groups could reduce the concentration of MDA in hepatic homogenate and raise the activation of SOD,GSH-PX,the concentration of GSH in liver tissue（P<0.01 or P<0.05）,Different doses of SZZJYA groups could alleviate liver injury（P<0.01 or P<0.05）.Immunohistochemical result showed that different doses of SZZJYA could increase the expression of TNF-αand iNOS（P<0.01）.Conclusion:（1）SZZJYA shows protective effect on CCl₄ induced chemical hepatic injury in mice.The mechanism may relate to anti-lipid peroxidation,attenuating oxide free radical.（2）SZZJYA shows protective effect on D-GalN induced chemical hepatic injury in mice.The mechanism may relate to repairing protein and DNA synthesis and anti-apoptosis.（3）SZZJYA shows protective effect on APAP induced drug-induced hepatic injury in mice.The mechanism may relate to the inhibition of cytokine expression,and the regeneration of liver cells.