摘要
目的应用分子生物学技术探讨黄芪三萜皂苷(total extract of astragalus,TEA)对柯萨奇B组3型病毒(CVB3)致细胞凋亡的保护作用。方法利用CVB3感染Vero细胞建立细胞凋亡模型并使用TEA对模型进行治疗,检测DNA Ladder以了解凋亡细胞DNA断裂情况;通过TUNNEL标记法、Annexin-V/PI染色、Hoechst33258染色观察细胞凋亡程度,比较各组间细胞凋亡的差异。结果 CVB3感染后,Vero细胞DNA的紫外成像呈现阶梯状条带,TEA组可观察到DNA断裂程度降低;TUNNEL检测可看到凋亡存在,病毒组显色较深,细胞凋亡程度较TEA组严重;Annexin-V/PI染色可见病毒组Vero细胞膜呈绿色荧光,细胞核显红色,核凝聚及裂解,并有凋亡小体存在,TEA组细胞状态改善;Hoechst33258染色结果表明,病毒组Vero细胞核显蓝色荧光,可看到凋亡中期核染色质凝聚、边缘化及晚期凋亡小体的典型变化。TEA组细胞状态改善,凋亡细胞数目减少。正常细胞对照组的细胞贴壁状态良好。流式细胞仪检测结果表明,黄芪三萜皂苷能够显著的降低试验组Vero细胞的凋亡率。结论 TEA对CVB3病毒感染的Vero细胞具有一定的保护作用,抑制细胞凋亡,其作用机制尚需做进一步探讨。
Objective To investigate the protective effect of total extract astragalus（TEA） on apoptosis of Vero cells caused by CVB3.Methods Vero cells were infected by CVB3 to establish an apoptosis model.DNA damage and apoptosis of model cells were detected by DNA ladder,TUNEL,Annexin-V/PI assay and Hoechst33258 assay to evaluate the therapeutic effect of TEA.Results DNA ladder assay showed that TEA decreased the DNA damage in the model group.The color of the model group was rather deep in TUNNEL assay,indicating the apoptosis in the model group was more severe than TEA groups;The cell membrane and cell nuclei showed green and red fluorescence respectively in Annexin-V/PI assay,and the model group displayed apoptotic body and condensed nucleoli,which were improved in the TEA groups.Hoechst33258 assay showed that cell nuclei showed blue fluorescence,condensed chromosomal in the model group and marginalisation,which were a typical change in early apoptotic bodies.Flow cytometry showed that TEA sharply decreased apoptotic percentage of Vero cells infected by CVB3.Conclusion TEA can inhibit the apoptosis of Vero cells infected by CVB3,but the mechanism remains unclear and requires further research.
出处
《中南药学》
CAS
2012年第10期730-734,共5页
Central South Pharmacy
基金
国家教育部"春晖计划"项目(编号:Z2004-2-15001)
内蒙古自治区自然科学基金资助项目(编号:200408020908)
内蒙古自治区科技计划项目(编号:20010214
20061108)
