| PART I The protection of the corilagin in treatment of myocardial inflammatory response of mice with Viral Myocarditis.Objective: To explore the protection of the crude extracts from the Heatclearing and Detoxifying Herbs corilagin in treatment of myocardial inflammatory response of the mice with viral myocarditis induced by Coxsackie virus B3.Methods: Totally 36 healthy male Balb/c mice were randomly divided into 6 group,including the control group,the model group,the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/k g),with 6 mice in each group.All of the groups were induced by intraperitoneal injecting with 0.1ml coxsackie group B type3(CVB3)virus to establish viral myocarditis mouse models except for the control group.Within 2 hours of virus induction,the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)were respectively treated by intraperitoneal injecting corilagin or dexamethasone,the other two groups were given Culture medium by intraperitoneal injection once daily for 7 days.The expression levels of aspartate aminotransferases(AST),lactate dehydrogenase(LDH),creatine kinase(CK),creatine kinase isoenzyme MB(CK-MB)and cardiac troponin I(c Tn I)were detected by using automatic biochemical analyzer.Then,their degree of myocardial pathological damage observed by HE staining.Results:1.The expression levels of AST,LDH,CK,CK-MB and c Tn I of the model group,the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)were significantly higher than those of the control group(P<0.05);The expression levels of AST,LDH,CK,CK-MB and c Tn I of the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)were significantly lower than those of the model group(P<0.05),In addition,the expression levels of myocardial zymogram were negatively correlated with the dose of corilagin.2.The pathological changes of myocardial tissue of mice in each group were observed by light microscope:Compared with the control group,the myocardial tissue of mice in the model group was seriously damaged,including the rupture of myocardial fibers,inflammatory cell infiltration,the Nuclear aggregation and unclear boundary of cardiomyocytes;Compared with the model group,the improvement effect of the damage of myocardial cell was positively correlated with the dose of corilagin and there was no significant difference between the high-dose corilagin group(40mg/kg)and the dexamethasone group.Conclusion(s): The crude extracts from the Heat-clearing and Detoxifying Herbs corilagin can reduce myocardial injury in VMC mice by decreasing the expression levels of Myocardial zymogram and reducing inflammatory reaction,and It was a protective effect on the inflammatory injury of myocardial tissue in mice with viral myocarditis induced by Coxsackie virus B3.PART II The intervention effect of the corilagin on TLR3 signaling pathway in myocardium of mice with Viral Myocarditis.Objective:To explore the intervention effect of the crude extracts from the Heat-clearing and Detoxifying Herbs corilagin on TLR3 signaling pathway in myocardial tissue of the mice with viral myocarditis induced by Coxsackie virus B3,and to explore the possible mechanism of corilagin intervention.Methods: Totally 36 healthy male Balb/c mice were randomly divided into 6 group,including the control group,the model group,the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/k g),with 6 mice in each group.All of the groups were induced by intraperitoneal injecting with 0.1ml coxsackie group B type3(CVB3)virus to establish viral myocarditis mouse models except for the control group.Within 2 hours of virus induction,the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)were respectively treated by intraperitoneal injecting corilagin or dexamethasone,the other two groups were given Culture medium by intraperitoneal injection once daily for 7 days.The m RNA levels of TLR3 、 TRIF、 TRAF6 and NEMO were measured by real-time fluorescence quantitative PCR(Real-time PCR);the protein levels of TLR3、TRIF、TRAF6 and NEMO were detected by Western blot,and the nuclear transcription levels of NF-κB from myocardial tissue of mice was detected by electrophoretic mobility shift assay(EMSA).Results:1.The results that the protein levels were detected by Western blot showed that the protein levels of TLR3、TRIF、TRAF6 and NEMO of the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)were significantly higher than those of the control group and the model group(P<0.05);the protein levels of TRAF6 and NEMO of the dexamethasone group(1mg/kg)were significantly lower than those of the high-dose corilagin group(40mg/kg)(P<0.05);and the protein levels of TRIIF was no significant differences between the high-dose corilagin group(40mg/k g)and the dexamethasone group(1mg/kg)(P>0.05).2.The results that the m RNA levels were measured by real-time fluorescence quantitative PCR(Real-time PCR)showed that the m RNA levels of TLR3、TRIF、TRAF6 and NEMO of the low-dose corilagin group(10mg/kg),the middle-dose corilagin group(20mg/kg),the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)were significantly higher than those of the control group and the model group(P<0.05),in addition,the m RNA levels of myocardial zymogram were positively correlated with the dose of corilagin;the m RNA levels of TRAF6 and NEMO of the dexamethasone group(1mg/kg)were significantly lower than those of the high-dose corilagin group(40mg/kg)(P<0.05);and the m RNA levels of TRIF was no significant differences between the high-dose corilagin group(40mg/kg)and the dexamethasone group(1mg/kg)(P>0.05).3.The results that the nuclear transcription levels of NF-κB was detected by electrophoretic mobility shift assay(EMSA)showed that the model group,the low-dose corilagin group(10mg/kg),the middle-do se corilagin group(20mg/kg),the high-dose corilagin group(40 mg/kg)and the dexamethasone group(1mg/kg)were significantly higher than those of the control group(P<0.05),and the nuclear transcription levels of NF-κB of the dexamethasone group(1mg/kg)were significantly lower than those of the high-dose corilagin group(40mg/kg)(P <0.05).Conclusion(s): 1.The crude extracts from the Heat-clearing and Detoxifying Herbs corilagin can reduce myocardial injury in VMC mice by inhibiting the activity of NF-κB and decreasing the expression of proinflammatory cytokines and It was a protective effect on myocardium in mice.2.Corilagin may have a synergistic effect in the signaling cascade of TLR3 signaling pathway,and it can promote the expression levels of TLR3、TRIF、TRAF6 and NEMO by up-regulating the expressions of TLR3. |
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