| ObjectiveJie Du Hua Yu Tang(JDHYT),as a compound of traditional Chinese herbs,are mainly composed of Isodon Lophanthoides var.gerardianus(Benth.) Hara,Oldenlandia diffusa(willd.) Roxb,Pora cocos(Schw.) Wolf,Hypericum japonicum rhumb,ex Murray,Paeonia veitchii lynch,Polygonum cuspidatum sieb.et Zucc and Glycyrrhiza uralensis Fish,have the function of clearing away the heat-evil,expelling superficial evils,promoting blood circulation and removing blood stasis,has been used for treating viral hepatitis with symptoms and signs of wetness-heat of liver and gallbladder,the therapeutic effect of anti-liver injury has been confirmed.To study the anti-experimental immune liver injury action and mechanism of JDHYT,We set up the animal model of Immune Liver Injury induced by Concanavalin A,to observe the effect on liver injury index in serum,hepatic histopathological change and the secretion of immune factors,to provide the pharmacology proof of JDHYT that be utilized in clinical treatment.Experimental contents1.Protective Effects of JDHYT on immune Hepatic Injury in Mice Induced by Concanavalin ASeventy-two male NIH mice were randomly divided into six groups:normal control group(group A),model group(group B),high dosage group of JDHYT(group C),middle dosage group of JDHYT(group D),low dosage group of JDHYT(group E) and bifendate group(group F).Excepts those in group A,mice in other groups were injected with Con A(dose of 20mg/kg) via the tail vein at the first day of experiment.Group C,Group D and group E were orally administered with JDHYT at does dose of 72.0g/kg,36.0g/kg and 18.0g/kg respectively.GroupF were orally administered with bifendate at the does dose of 150mg/kg.All the drugs were given once daily for 5 days consecutively.At the last time,when drugs were administered for 2 hours,mice were injected with Con A once again at the same dosage.Blood sample for determineing aminotransferase(ALT) and aspartate aminotransferase(AST) levels were collected at the 8th hour after Con A administration.Histopathological examination was performed for liver tissue.It showed that the serum levels of ALT and AST in all groups of JDHYT were obviously lower to that of group B(P＜0.01),Serum levels of ALT and AST in group C were obviously lower to that of group E(P＜0.01).Serum levels of ALT in all groups of JDHYT have no significant difference comparing with that of group F(P＞0.05),but serum level of AST in group C were obviously lower to that of group F(P＜0.05).hepatic histopathological change of necrosis were alleviated in all groups of JDHYT.It suggested that JDHYT possess the effects of protecting liver from immune injury in mice induced by Concanavalin A.2.Anti-Experimental immune liver injury action and mechanism of JDHYT in mice induced by Concanavalin A2.1 Effects of JDHYT on the expression of SOD,MDA,ICAM-1,Fas,FasL and IL-6 in immune liver injury mice induced by Concanavalin A.Sixty male NIH mice were randomly divided into 5 groups:normal control group(group A),model group(group B),high dosagegroup of JDHYT(group C), low dosage group of JDHYT(group D) and bifendate group(group E).Excepts those in group A,mice in other groups were injected with Con A(dose of 20mg/kg) via the tail vein at the first day of experiment.Group C and group D were orally administered with JDHYT at does dose of 72.0g/kg and 18.0g/kg, respectively.Group E were orally administered with bifendate at the does dose of 150mg/kg.All the drugs were given once daily for 5 days consecutively.At the last time,when drugs were administered for 2 hours,mice were injected with Con A once again at the same dosage,then the blood samples and liver tissue were collected for determining the level of ALT,IL-6,superoxide dismutase(SOD) and maiondiadehyde(MDA),the expression of ICAM-1,Fas and FasL were detected by immunohisttochemical method.It showed that the level of ALT and IL-6 in group C and group D were obviously lower to that of group B(P＜0.01,0.05),the level of ALT in group C was obviously lower to that of group D(P＜0.01),but no significant difference was observed comparing with that of group E(P＞0.05).The activity of SOD in liver tissue in group C and group D were obviously higher to that of group B(P＜0.01,0.05),but no significant difference was observed comparing with that of group E(P＞0.05),the activity of SOD in liver tissue in group C was obviously higher to that of group D(P＜0.05)The level of MDA in liver tissue in group C and group D were obviously lower to that of group B(P＜0.01),the level of MDA in group C was obviously lower to that of group D(P＜0.01),but no significant difference was observed comparing with that of group E(P＞0.05).Hepatic histopathological changes were alleviated in group C and group D comparing with that in group B(P＜0.05),the experssion of ICAM-1,Fas and FasL in liver tissue of group C and group D were obviously attenuated comparing with that of group B(P＜0.01,0.05).It suggested that JDHYT has a good effects of protecting liver from immune injury in mice by antioxidation and suppressing the expression of ICAM-1, Fas,FasL and IL-6.2.2 Effects of JDHYT on the production of TNF-αand the change of ultrastructure of liver cells in mice induced by Concanavalin AFifty male NIH mice were randomly divided into 5 groups:normal control group(group A),model group(group 8),high dosage group of JDHYT(group C), low dosage group of JDHYT(group D) and bifendate group(group E).Excepts those in group A,mice in other groups were injected with Con A(dose of 20mg/kg) via the tail vein at the first day of experiment.Group C and group D were orally administered with JDHYT at does dose of 72.0g/kg and 18.0g/kg, respectively.Group E were orally administered with bifendate at the does dose of 150mg/kg.All the drugs were given once daily for 5 days consecutively.In the 5th day,after the drugs were administered for 2 hours,mice were injected with Con A once again at the same dosage.After 4 hours,the blood of eight mice in all groups was obtained and the serum was separated fasting.After 8 hours,the liver tissue of other two mice in all groups were get to observe the change of ultrastructure of hepatocyte under electron microscope.It showed that contents of TNF-αin serum in group C and group D were obviously lower to that of group B(P＜0.01),Histopathological change of ultrastructure of hepatocyte were alleviated in both groups of JDHYT.It suggested that JDHYT possess the effects of protecting liver from immune injury in mice induced by Concanavalin A by suppressing the production of TNF-α.Conclusion1.JDHYT possess the effects of protecting liver from immune injury in mice induced by Concanavalin A.2.JDHYT has a good effects of protecting liver from immune injury in mice by antioxidation,suppressing the expression of ICAM-1,Fas and FasL,and by inhibiting the secretion of IL-6.3.JDHYT possess the effects of protecting liver from immune injury in mice induced by Concanavalin A by suppressing the production of TNF-α. |
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