血必净对活化诱导T细胞凋亡的调节

目的 观察活化诱导对脾脏T淋巴细胞凋亡、凋亡相关基因mRNA表达及caspase3活性的影响,以及活血化瘀中药的调节作用.方法 提取BALB/c小鼠脾脏T淋巴细胞并培养,以Con A+IL-2诱导T细胞活化凋亡,MTT法检测细胞增殖活性,流式细胞仪检测细胞凋亡率,RT-PCR检测Fas、FasL、Bcl-2、Bax、IL-2 mRNA表达水平,分光光度法测定caspase3酶活性,并观察活血化瘀中药对上述各项指标的影响.结果 活化T淋巴细胞于诱导18h后凋亡率明显增加,于诱导6h时未见FasL、Bax mRNA表达,Fas、Bcl-2 mRNA表达无明显变化;于诱导18 h后Fas、FasL、Bax mRNA表达升高,Bel-2 mRNA表达下降,caspase3活性增高.活血化瘀中药可降低T细胞凋亡,并可分别降低Fas、FasL、Bax mRNA表达,提高Bcl-2 mRNA表达,减轻easpase3酶活性.在活化诱导早期(6 h)促进T淋巴细胞内IL-2 mRNA表达,在晚期(18 h)减少IL-2 mRNA表达.结论 过度活化是脾脏T淋巴细胞异常凋亡的诱发因素,而凋亡的发生与Fas、FasL、Bax、Bcl-2 mRNA表达的改变有关.活血化瘀中药可通过调节IL-2及凋亡相关基因mRNA表达而减轻脾脏T淋巴细胞凋亡,同时可以促进T淋巴细胞的增殖活性.

The effect of Xuebijing on the activation-induced cell death (AICD) of T lymphocyte

Objective To observe the influence of Xuebijing injection on apoptosis, apoptotic related gene mRNA levels and activity of caspase3 in activated T lymphocyte. Methods The T lymphocytes were obtained from the spleens of BALB/c mice and be induced to be activated and apoptotic by cultured with Con A + IL-2. Apoptosis was investigated by flow cytometry. RT-PCR was used to detect the expression of Fas, FasL, Bcl-2, Bax, IL-2 mRNA, and the activity of caspase3 in T lymphocyte was also detected by spectrophotometric method. In the mean time, the effect of Xuebijing injection on those parameters was observed. Results After the induction, T lymphocyte apoptosis raised at 18 h. At 6 h after the induction, there was no expression of FasL, Bax mRNA, and no change in the expression of Fas and Bcl-2 mRNA. At 18 h, the expressions of Fas, FasL, Bax mRNA rised and the expression of Bcl-2 mRNA lessened. The activity of caspase3 also ascended. Xuebijing injection can cut down the apoptosis induced by induction, make the expression of Fas, FasL, Bax mRNA decreased and Bcl-2 mRNA improved. The activity of caspase3 also fallen after the Xuebijing injection treated. It can promote the expressions of IL-2 mRNA at early phase of AICD (6 h) and depress the expressions at the late period (18 h). Conclusion The apoptosis of T lymphocyte induced by activation was regulated by the change of Fas, FasL, Bcl-2, Bax mRNA expression. Xuebijing injection can ameliorate the apoptosis through regulating the expression of IL-2 and apoptotic related gene mRNA, improve the proliferation activity of T lymphocyte.