Study On The Relationship Of IL-18 With Immuno-pathological Lesion And The Regulation Of Chinese Herbs In Severe Acute Pancreatitis

PartⅠThe Relationship of Serum IL-18 with Immuno-Pathological Changes in Rats of Severe Acute PancreatitisObjective:To investigate the relationship of serum IL-18 with IL-1β,to study the relationship of serum IL-18 with splenic lymphocyte apoptosis rate and the expression of FasL and Bcl-2 mRNA of splenic lymphocyte,to analyze the relationship of serum IL- 18 with immuno-pathological changes in rats of severe acute pancreatitis.Methods:16 healthy Wistar rats were randomly divided into two teams:control group(6 rats)and SAP group(10 rats).SAP model was induced by retrograde infusion of 5%Sodium Cholate into the bile-pancreatic duct.Death rate of each group was recorded at 24 hour after the operation.Blood of all rats was taken from femoral artery to detect the levels of serum IL-18 and IL-1βat 24 hour after the operation. Lymphocytes were drawn off from spleen to detect apoptosis rate using flow cytometer and the expression of FasL and Bcl-2 mRNA by semi-quantitative RT-PCR.Pancreas,lung,liver and kidney were stored to perform histopathologic examination.Results:The levels of serum IL-18 and IL-1βand the pathological scores in SAP group were significantly higher than that in control group(P＜0.01),the levels of serum IL-18 had positive correlation with the levels of serum IL-1βand the pathological scores(r=0.886 and r=0.807,P＜0.01).The apoptosis rates of splenic lymphocyte and the expression of FasL mRNA in SAP group were significantly higher than that in control group(P＜0.01),while the expression of Bcl-2 mRNA in SAP group was significantly lower than that in control group(P＜0.01).In SAP group, the levels of serum IL-18 had positive correlation with the apoptosis rates of splenic lymphocyte and the expression of FasL mRNA(r=0.650 and r=0.713,P＜0.05),while the levels of serum IL-18 had negative correlation with the expression of Bcl-2 mRNA(r=0.836,P＜0.01).Conelusion:Both IL-18 and IL-1βby reciprocal induction participate systemic inflammation response and then aggravate the pathological lesion of multiple organ during severe acute pancreatitis;There is lymphocyte apoptosis increasing in the early stage of SAP,which may an important reason of immunosuppression in the early stage of SAP;An important pathway of splenic lymphocyte apoptosis increasing may be the expression increasing of FasL mRNA and the expression decreasing of Bcl-2 mRNA;IL-18 may effect on lymphocyte apoptosis increasing by up-regulating the expression of FasL mRNA and down-regulating the expression of Bcl-2 mRNA. PartⅡThe Regulation of Chinese Herbs to Immuno-Pathological Changes in Rats of Severe Acute PancreatitisObjective:To investigate the effect of LaiFuChengQiTang and ShenQiFuZhengZhuSheYe on serum cytokines,apoptosis rates of splenic lymphocyte,apoptosis related genes and multiple organ pathological changes;to analyze the regulation of these two Chinese Herbs to immuno-pathological changes in rats of severe acute pancreatitis.Methods:36 healthy Wistar rats were randomly divided into four teams:control group(K group,6 rats),SAP group(M group,10 rats),LaiFuChengQiTang group(L group,10rats)and ShenQiFuZhengZhuSheYe group(S group,10 rats).SAP model was induced by retrograde infusion of 5%Sodium Cholate into the bile-pancreatic duct in M group,L group and S group.In L and S groups,all rats were treated by intragastric administration of homologous Chinese herbs(1.25ml/100g)at 8 hour and 16 hour after operation,while in K and M groups,all rats were treated by intragastric administration of normal sodium(1.25ml/100g)at 8 hour and 16 hour after operation. Death rate of each group was recorded at 24 hour after the operation.Blood of all rats was taken from femoral artery to detect the levels of serum IL-18 and IL-1βat 24 hour after the operation.Lymphocytes were drawn off from spleen to detect apoptosis rate using flow cytometer and the expression of FasL and Bcl-2 mRNA by semiquantitative RT-PCR.Pancreas,lung,liver and kidney were stored to perform histopathologic examination.Results:The levels of serum IL-18 and IL-1β,the multiple organ pathological scores,the apoptosis rates of splenic lymphocyte and the expression of FasL mRNA in LaiFuChengQiTang group were lower than that in SAP group(P＜0.05 or P＜0.01) but remain higher than that in control group(P＜0.01),while the expression of Bcl-2 mRNA was higher than that in SAP group(P＜0.01)but remain lower than that in control group(P＜0.01).The levels of serum IL-18 and IL-1β,the multiple organ pathological scores,the apoptosis rates of splenic lymphocyte and the expression of FasL mRNA in ShenQiFuZhengZhuSheYe group were lower than that in SAP group (P＜0.01)but remain higher than that in control group(P＜0.05 or P＜0.01),while the expression of Bcl-2 mRNA was higher than that in SAP group(P＜0.01)but remain lower than that in control group(P＜<0.05).Conclusion:LaiFuChengQiTang can down-regulate the expression of serum IL-18 and IL-1βand significantly lessen multiple organ pathological lesion,which confirm the therapeutic effect of LaiFuChengQiTang on SAP.LaiFuChengQiTang can downregulate the expression of FasL mRNA and up-regulate the expression of Bcl-2 mRNA,which may be an important pathway of LaiFuChengQiTang to reduce lymphocyte apoptosis.ShenQiFuZhengZhuSheYe used in the early stage of SAP also can down-regulate the expression of serum IL-18 and IL-1βand lessen multiple organ pathological lesion,which tentative confirm the therapeutic effect of ShenQiFuZheng ZhuSheYe on SAP.ShenQiFuZhengZhuSheYe also can reduce lymphocyte apoptosis, which may come true by down-regulating the expression of FasL mRNA and upregulating the expression of Bcl-2 mRNA.But the effect of LaiFuChengQiTang and ShenQiFuZhengZhuSheYe on the immune function of SAP should be confirmed in clinical therapy.