The Mechanism Of Bushen Jiedu Decoction Against Immunological Liver Injury In Mice

Objective: To study the protective role of Bushen Jiedu decoction,disassembled decoction and single decoction dry extract by Con A-induced immunological liver injury in mice;To study the effect of the above herbal on the proliferation of human LO2 cell and splenocytes;To study the effect of the above herbal on the levels of cytotoxic in the primary splenic lymphocytes and peritoneal macrophages in mice.In order to explain the immune regulation mechanism of Bushen Jiedu decoction,which protects the liver injury.Method: 1.Establish a model of immunological liver injury with Con A in KM mice;2.Bushen Jiedu decoction was used to intervene in mice of the Con A model that observed liver histopathology by HE method,and detected the levels of ALT,AST in serum and SOD,MDA,CAT in liver homogenate supernatant by biochemical method,and detected the levels of IFN-? in splenocytes supernatant by ELISA.In order to study the protective effect of Bushen Jiedu Decoction on liver injury;3.Bushen Jiedu decoction was used to intervene in Human liver LO2 cells that tested its proliferation by MTT,in order to study the the regulation of Bushen Jiedu Decoction on hepatocytes proliferation;4.Bushen Jiedu decoction was used to intervene in the primary splenocytes and peritoneal macrophages in mice that tested its proliferation by MTT,and the levels of IFN-? in splenic lymphocytes supernatant and TNF-? in peritoneal macrophages supernatant.In order to study the regulation of Bushen Jiedu Decoction on immune cells.Result: 1.The protective role of Bushen Jiedu decoction on immunological liver injury in mice:(1)the effects of herbals on organ coefficient in model mouse: compared with the normal group,the index of liver significantly rises up in model group(P<0.05),and the index of thymus was decreased(P>0.05);compared with the model group,the whole decoction,Huoxue Jiedu decoction and Citrus Reticulate group significantly reduce the elevated liver index(P<0.05);(2)the effects of herbals on Pathology in mice:compared with the normal group,pathology showed obvious hepatocyte necrosis area with a large number of inflammatory cells aggregation and fibrocytes proliferation in the model group;compared with the model group,Pathology of treatment group showed that the hepatocyte necrosis area and inflammatory cell aggregation was reduced,binuclear or multinuclear hepatocytes appeared simultaneously;(3)the effects of herbals on liver function in mice: compared with the normal group,ALT and AST of the serum were significantly increased in the model group(P<0.05);compared with the model group,Huoxue Jiedu decoction group reduced the level of serum AST,Morinda Officinalis,Epimedium Brevicornu and Sophora Flavescentis group reduced the level of serum ALT(P<0.05);(4)the effects of herbals on liver antioxidant ability in mice: compared with the normal group,SOD and CAT of the liver homogenate supernatant showed a decreased trend in the model group(P>0.05);compared with the model group,Sophora Flavescentis,Salvia Miltiorrhiza,Ranunculus Ternatus,Citrus Reticulata group elevate the activity of SOD and CAT in liver tissues(P<0.05);(5)the effects of herbals on the expression levels of IFN-? in mice: compared with the normal group,the expression levels of IFN-? was increased in the model group(P<0.05);compared with the model group,the whole decoction one group,Bushen decoction group,Morinda Officinalis,Sophora Flavescentis group decreased the expression levels of IFN-?(P<0.05);2.The effects of Bushen Jiedu Decoction on human liver LO2 cells:(1)The whole decoction and three kinds of disassembled decoction have obvious proliferative activity in 10,50,100 and 250?g/m L(P<0.05),and they have dose-effect relationship;(2)250?g/m L Epimedium Brevicornu,Rehmannia Glutinosa and Ganoderma Lucidum has a proliferative effect(P<0.05);50,100 and 250?g/m L Salvia Miltiorrhiza and Citrus Reticulata has a proliferative effect(P<0.05);(3)50,100 and 250 ?g/m L whole decoction dry extract had an obvious inhibitory effect(P<0.05),and showed dose-effect relationship;(4)250 ?g/m L Bushen and Huoxue Jiedu dry extracts showed an obvious regulating effect on suppressing proliferation by H2O2(P<0.05);50?g/m L Salvia Miltiorrhiza and 100,250?g / m L Citrus Reticulata showed the regulating effect on suppressing proliferation by H2O2(P <0.05);3.The effects of Bushen Jiedu Decoction on immune cells:(1)The whole decoction and three disassembled decoction dry extracts have no obvious proliferative effect on splenocytes and antagonistic effect on Con Astimulated and LPS-stimulated splenocytes proliferation(P<0.05);(2)The whole decoction has no effect on the secretion of IFN-? in primary splenocytes,but it can reduce the expression level of TNF-? in peritoneal macrophages;(3)250?g/m L Bushen decoction,Jianpi decoction,Huoxue Jiedu decoction promoted the expression of IFN-? in splenic lymphocytes(P<0.05)and inhibited the expression of TNF-? in peritoneal macrophages.Conclusion: 1.Bushen Jiedu Decoction has therapeutic effects on mice with immunological liver injury mainly by reducing the levels of transaminase,increasing the activity of liver superoxide dismutase,and the effect of whole decoction is better than disassembled decoction and single decoction.2.The whole decoction,Bushen decoction,Jianpi decoction,Huoxue Jiedu decoction,Epimedium Brevicornu,Rehmannia Glutinosa,Ganoderma Lucidum,Salvia Miltiorrhiza,and Citrus Reticulata can promote the proliferation of hepatocytes and inhibit the oxidative damage and apoptosis of hepatocytes induced by H2O2.3.Bushen Jiedu Decoction regulates the immune function of primary splenic lymphocytes and peritoneal macrophages in mice.