雷公藤甲素对C6胶质瘤细胞凋亡及TNF-α、NF-κb和Caspase-3表达的影响

Triptolide inhibits proliferation and induces apoptosis of C6 cells by regulating the expressions of TNF-α、NF-κb and Caspase-3

目的:检测雷公藤甲素对C6胶质瘤细胞TNF-α、NF-κb和Caspase-3表达的影响,探讨其诱导胶质瘤细胞凋亡机制。方法:将大鼠C6胶质瘤细胞培养于含10%FBS,100 mg/L青霉素链霉素的DMEM培养液中,设立雷公藤甲素给药组和空白组,给药组设0.2、0.4、0.8、1.6、3.2、6.4μg/L雷公藤甲素6个剂量,置37℃5%CO2培养箱中培养,采用四甲基偶氮唑盐比色(MTT)法检测雷公藤甲素对大鼠C6胶质瘤细胞的抑制作用,酶联免疫(Enzyme Linked Immunosorbent Assay,ELISA)试剂盒检测TNF-α表达,Western Blotting(WB)检测NF-κb和Caspase-3的表达。结果:与空白组相比,浓度大于0.8μg/L的雷公藤甲素能诱导C6胶质瘤细胞凋亡(P<0.05),且其诱导效应具有浓度依赖性。同时,雷公藤甲素可以下调NF-κb表达,而上调TNF-α和Caspase-3表达,且Caspase-3被部分活化。结论:雷公藤甲素可诱导C6胶质瘤细胞凋亡,同时,可降低NF-κb表达,增强TNF-α和Caspase-3表达,且部分活化Caspase-3。

Objective ：To investigate the effect of triptolide on the expressions of TNF-α、NF-κb and Caspase-3 on C6 glioma cells. Methods： C6 glioma cells was cultivated in vitro and treated with different concentrations of triptolide as 0.2,0.4,0.8,1.6,3.2,6.4 μg/L, and blank group was established as control, cell viability was determined by MTr assay. The expression of TNF-α was determined by ELISA Kit while NF-Kb and Caspase-3 by Western Blotting. Results Compared with the control group, apoptosis was induced obviously by triptolide above 0. 8μg/L, with concentration dependent. With elevating concentration of triptolide, the expression of NF-κb was down regulation while the tendency of TNF-α expression was opposite, and Caspase-3 was expression up regulation, with partial activation. Conclusion： Triptolide induced apoptosis of C6 glioma cells, TNF-α, NF-Kb and Caspase-3 may be involved in the apoptosis of C6 glioma cells.