芪红胶囊对柯萨奇病毒引起细胞凋亡的影响

Effect of Qihong Capsule in Inhibiting Cell Apoptosis Induced by Coxsackie Virus B

目的探讨柯萨奇病毒是否能够引起心肌细胞凋亡,以及芪红胶囊对凋亡发生是否有抑制作用。方法将培养细胞分为4组:病毒感染对照组、芪红胶囊加病毒组、芪红胶囊对照组和正常细胞对照组。应用原位末端标记法(TUNEL)、Hoechst33258染色和Annexin-Ⅴ/PI染色观察各组细胞凋亡发生情况;并通过流式细胞仪分析各组细胞的凋亡发生率;应用RT-PCR方法观察与凋亡相关细胞因子的表达改变。结果病毒感染对照组细胞经Hoechst33258染色发现病毒感染后的细胞核出现强亮度的蓝色荧光,可观察到凋亡细胞的典型变化;Annexin-Ⅴ/PI染色可见HeLa细胞膜呈强绿色荧光,细胞核显强红色荧光;通过流式细胞仪检测PI染色的细胞DNA,发现病毒感染组出现明显凋亡峰;芪红胶囊加病毒组凋亡发生率明显下降,正常对照组细胞未发生凋亡;同时芪红胶囊能够调节与凋亡相关细胞因子的表达。结论芪红胶囊能够抑制柯萨奇病毒引起的细胞凋亡。

ObjectiveTo investigate whether the cell apoptosis could be induced by Coxsackie virus B (Cox B) and Qihong capsule (QHC) has the inhibition on the cell apoptosis. MethodsCultured cells were divided into 4 groups, the Cox B infected group, the QHC treated and the Cox B infected group, the QHC control group and the normal control group. The cells apoptosis was determined by TUNEL labeled in situ, Hoechst 33258 staining and Annexin-V/PI staining, the apoptotic incidence was assayed by flow cytometry, and the change in expression of apoptotic related cytokines was measured with RT-PCR. ResultsThe Cox B infected cell nucleus displayed strong blue fluorescence by Hoechst 33258 staining, and typical change of apoptotic cells could be detected. HeLa cell membrane showed strong green fluorescence and nucleus showed strong red fluorescence by Annexin-V/PI staining. Flow cytometric observation on DNA of PI stained cells showed obviously an apoptotic peak in the Cox B infected group. QHC could decrease the apoptosis incidence, while there was no cell apoptosis occurred in the normal control group. Besides, QHC could regulate the expression of apoptotic related cytokines. ConclusionQHC has effect in inhibiting cell apoptosis induced by Cox B.