| Objective:To explore the protective effect of Xinan granule on viral myocarditis mice and its possible molecular mechanisms basing on the nuclear factor-?b(NF-?B)signal pathway and cell apoptosis.Methods:120 BALB/c mices according to random number table method is divided into the normal group,the model group,the ribavirin group,the Xinan granule high,medium and low dose group,20 mices of each group.The mice in model and each medic ine groups were inoculated with 0.2ml cultures containing 103TCID50 Coxsackievirus B3(CVB3)intraperitoneally;and those in normal control group were inoculated with 0.2mL cultures without CVB3.After the mices were inoculated CVB3 2h later,administrating the ribavirin group with ribavirin 0.2mg/(g·d),The Xinan granule high,medium and low dose group respectively with Xinan granules 10,20,40g/(kg·d),the normal group and model group with the same amount of pure water for 7days.The mice were killed on day 8 after weighing body mass and taking blood by removalling eyeball.Serum was separated.Serum c TnI and NO concentration were detected using ELISA.The heart was quickly removed and weighed to calculate heart index.Histological sections of heart were stained with HE and myocardial histopathologic scores were counted under microscope.The expression of NF-?B p65,P-I?Ba and Caspase-3 were examined by Western blotting.The expression level of TNF-?,IL-6,IFN-?and CD68 positive macrophage were detected by immunohistochemistry staining.Results: 1)mortality of mice:There were no death cases in normal group.The mortality of Xinan granule high,medium dose group and ribavirin group decreased significantly(P<0.01)to compare with model group.There was no statistical difference between low dose group and model group;Compared with low dose group,the mortality of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with ribavirin group,the mortality of medium dose group decreased,high dose group had no obvious change,but there were no statistical differences(P>0.05).2)HM/BM: Compared with normal group,the HM/BM of model group and each medic ine group increased(P<0.05);Compared with model group,the HM/BM of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with low dose group,the HM/BM of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with ribavirin group,the HM/BM of medium dose group had no obvious change,high dose group decreased,there were no statistical differences(P>0.05).3)Myocardial histopathologic scores:There were no obvious pathological changes in normal group.;Compared with model group,the histopathologic score of ribavirin group,medium,high dose group decreased significantly(P<0.01).The histopathologic score of low dose group had no obvious change(P>0.05);Compared with low dose group,the histopathologic score of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with ribavirin group,the histopathologic score of Xinan granule medium,high dose group increased,but there were no statistical differences(P>0.05).4)Expression level of CVB3 mRNA:There was no expression of CVB3 mRNA in normal group;Compared with model group,the CVB3 mRNA expression level of ribavirin group,medium,high dose group decreased significantly(P<0.01),low dose group had no obvious change(P>0.05);Compared with low dose group,the CVB3 mRNA expression level of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with ribavirin group,Xinan granule medium,high dose group decreased(P<0.05).5)Serum c TnI and NO detection:(1)Compared with normal group,the c TnI concentration of model group and each medic ine group increased(P<0.05);Compared with model group,the cTnI concentration of each medicine group decreased(P<0.05 or P<0.01);Compared with low dose group,the cTnI concentration of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with ribavirin group,Xinan granule medium,high dose group of c TnI concentration increased,but there were no statistical differences(P>0.05).(2)Compared with normal group,the NO concentration of model group and each medicine group increased(P<0.05);Compared with model group,the NO concentration of ribavirin group,medium,high dose group decreased significantly(P<0.01),but low dose group of NO concentration had no obvious change(P>0.05);Compared with Xinan granule low dose group,the NO concentration of ribavirin group,medium,high dose group decreased significantly(P<0.01);Compared with ribavirin group,the Xinan granule medium,high dose group had the trend to decrease,but with no statistical difference(P>0.05).6)TNF-?,IL-6,IFN-? protein and CD68 macrophages detection:(1)Compared with normal group,the TNF-? protein MOD of model group and each medic ine group increased(P<0.05);Compared with model group,the TNF-? protein MOD of ribavirin group,medium,high dose group decreased(P<0.05).But there was no statistical difference between low dose group and model group(P>0.05);Compared with Xinan granule low dose group,the TNF-? protein MOD of ribavirin group,medium,high dose group decreased(P<0.05);Compared with ribavirin group,the TNF-? protein MOD of high dose group slightly increased,but without statistical difference(P>0.05).(2)Compared with normal group,the IL-6 protein MOD of model group and each medic ine group increased(P<0.05);Compared with model group,the IL-6 protein MOD of ribavirin group,medium,high dose group decreased(P<0.05);Compared with Xinan granule low dose group,the IL-6 protein MOD of ribavirin group,medium,high dose group decreased(P<0.05);Compared with ribavirin group,the IL-6 protein MOD of high dose group had the trend to decrease,but with no statistical difference(P>0.05).(3)Compared with normal group,the IFN-? protein MOD of model group and each medic ine group increased(P<0.05);Compared with model group,the IFN-? protein MOD of ribavirin group,medium,high dose group decreased(P<0.05),low dose group slightly decreased,but there was no statistical difference(P>0.05);Compared with low dose group,the IFN-? protein MOD of ribavirin group,medium,high dose group decreased(P<0.05);Compared with ribavirin group,the IFN-? protein MOD of medium,high dose group had the trend to decrease,but there was no statistical difference(P>0.05).(4)There was no CD68 macrophage infiltration in normal group.Compared with model group,the CD68 macrophage numbers of ribavirin group,medium,high dose group decreased significantly(P<0.01).The CD68 macrophage numbers of low dose group slightly decreased,but without statistical difference(P>0.05);Compared with low dose group,the CD68 macrophage numbers of ribavirin group,medium,high dose group significantly decreased(P<0.01);Compared with ribavirin group,medium dose group of CD68 macrophage numbers increased(P<0.05).There was no statistical difference between high dose group and ribavirin group(P>0.05).7)NF-?Bp65,P-I?B? and Caspase-3 protein detection:(1)Compared with normal group,the NF-?B p65 protein expression of model group and each medic ine group increased(P<0.05);Compared with model group,the NF-?B p65 protein expression of ribavirin group,medium,high dose group decreased(P<0.01).But there was no statistical difference between low dose group and model group(P>0.05);Compared with low dose group,the NF-?B p65 protein expression of ribavirin group,medium,high dose group decreased(P<0.01);Compared with ribavirin group,the medium and high dose group of NF-?B p65 protein expression increased(P<0.05).(2)Compared with normal group,the model group,ribavirin group,low and medium dose group of P-I?B? protein expression increased(P<0.05);Compared with model group,the expression of P-I?B? protein in ribavirin group,medium,high dose group decreased(P<0.01).There was no statistical difference between low dose group and model group(P>0.05).Compared with low dose group,the P-I?B? protein expression of ribavirin group,medium,high dose group decreased(P<0.01);Compared with ribavirin group,the high dose group of P-I?B? protein expression decreased,but without statistical difference(P>0.05).(3)Compared with normal group,the model group,ribavirin group,low and medium dose group of Caspase-3 protein expression increased(P<0.05);Compared with model group,the expression of Caspase-3 protein in Xinan granule each dose group decreased(P<0.01);Compared with low dose group,the high dose group of Caspase-3 protein expression decreased(P<0.05),medium dose group had no significant change(P>0.05);Compared with ribavirin group,the Xinan granule each dose group of Caspase-3 protein expression significantly decreased(P<0.01).Conclusion: 1)The pathogenes is of VMC exist in the process of the NF-?B signal pathway activation and apoptosis.2)Xinan granule can obviously improve the myocardial inflammation and injury,and protect the myocardium of mice.3)The mechanisms of Xinan granule may be associated with the inhibition of NF-?B signal pathway activation and viral replication,adjusting expression levels of Caspase-3,and decreasing apoptosis signal conduction with function of inhibiting myocardial apoptosis.4)There has some quantity-effect relationship of Xinan granule on VMC mice.It shows a trend that is superior to the ribavirin on inhibition viral replication,reducing the NO concentration and the expression of IFN-? and Caspase-3 protein. |
PDF Full Text Request