目的：探讨柯萨奇病毒B3(Coxsackie virus B3,CVB3)对心肌微血管内皮细胞(Cardiac myocardial microvascular endothelial cell,CMEC)的感染途径及红花提取物(Safflower extract,SYB)是否通过该途径对感染过程起到保护作用；方法：将CMEC分为四组,分别为CVB3干预组、SYB干预组、CVB3+SYB干预组、空白对照组,采用TCID50浓度为10_^-6.285/ml浓度的病毒液对CVB3组及CVB3+SYB组的细胞进行干预,采用100nmol/L的SYB对SYB组及CVB3+SYB组的细胞进行干预,分别在0min、20min、40min、60min提取细胞,采用WB及RT-PCR方法,测定每组细胞的Toll-样受体4(Toll-like receptors 4,TLR4)、髓样分化因子88(Myeloid differentiation factor 88,MyD88)及核转录因子-kB(Nuclear factor-kappa B,NF-kB)条带灰度以及蛋白和mRNA含量,进行统计学分析；结果：在CVB3干预组与空白对照组相比,TLR4、MyD88及NF-kB的蛋白表达量在40min、60min存在差异,mRNA的表达量在20min、40min、60min时存在差异。SYB干预组与空白对照组相比,TLR4、MyD88及NF-kB在各时间段的蛋白表达量以及mRNA表达量均无差异。SYB+CVB3干预组与CVB3干预组对比,TLR4、MyD88及NF-kB蛋白表达量在40min与60min时存在差异,mRNA表达量在20min、40min、60min时存在差异；结论:CVB3可通过TLR4/NF-kB通路损伤CMEC,TLR4/NF-kB通路是CVB3导致VMC的信号通路,而SYB可通过该通路对CVB3感染后CMEC起到一定保护作用。

Objective: To discuss possible signaling pathways for CVB3 to damage CMEC through appropriate experimental methods and whether SYB might have a protective effect on CMEC through this signaling pathway. Methods: Culture CMEC and divide the cells into four groups: CVB3 intervention group, SYB intervention group, CVB3 + SYB intervention group and blank control group. The TCID50 concentration of 10^-6.285/ml CVB3 and the concentration of 100nmol/L SYB were used to intervene CVB3 group, SYB group and CVB3+SYB group. Extract the cells at the time of 0min,20min,40min and 60min.Use the methods of WB and RT-PCR to measure the protein and mRNA expression of TLR4, MyD88 and NF-kB. Results: Compared with the blank group, in the CVB3 intervention group, there were differences at 40min and 60min.The mRNA expression had differences at 20min,40min and 60min.The protein expression and mRNA expression of the SYB intervention group at each time period were no difference with the blank control group. Yet, the comparisons of protein expression between the SYB+CVB3 intervention group and the CVB3 intervention group had differences at 40min and 60min. The mRNA expression had differences at 20min,40min and 60min.Conclusions: CVB3 can damage the CMEC through TLR4/NF-kB pathway. TLR4/NF-kB pathway is the signal pathway of CVB3 which leads to viral myocarditis, while safflower extract has a certain protective effect on the CMEC after CVB3 infection through this way.

R542.2