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目的:探讨雷公藤甲素对大鼠C6胶质瘤细胞的增殖诱导作用及其相关机制。方法:将大鼠C6胶质瘤细胞培养于含10%(FBS),100 mg.L-1青霉素链霉素的DMEM培养液中,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,流式细胞仪(FCM)检测C6胶质瘤细胞凋亡情况,用逆转录-聚合酶链反应(Real Time-Polymerase Chain Reaction,RT-PCR),免疫印迹法(Western blotting,WB)检测细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax)mRNA及蛋白的表达变化情况。结果:1.2 mg.L-1浓度的雷公藤甲素溶液对C6胶质瘤细胞增殖有抑制作用,且呈明显的剂量依赖性;雷公藤甲素作用C6胶质瘤细胞24 h后,1.2 mg.L-1浓度对其凋亡诱导作用最为明显;雷公藤甲素可明显下调细胞中Bcl-2 mRNA及蛋白质表达,且该作用呈剂量-时间依赖性,而Bax mRNA及蛋白质表达呈上调趋势。结论:雷公藤甲素对C6胶质瘤细胞具有较好的生长抑制和凋亡诱导作用,而其作用机制可能与Bcl-2,Bax mRNA及蛋白质表达有关。
Objective: To investigate the effects of Triptolide on the proliferation of rat C6 glioma cells and its related mechanisms. METHODS: Rat C6 glioma cells were cultured in DMEM containing 10% (FBS) and 100 mg.L-1 penicillin streptomycin and cultured in a 37 ° C 5% CO 2 incubator. Tetramethyl The inhibitory effect of triptolide on C6 glioma cells was detected by MTT assay. The apoptosis of C6 glioma cells was detected by flow cytometry (FCM) (Bcl-2), Bcl-2-related protein B (Bax) mRNA and protein were detected by Real-Time Polymerase Chain Reaction (RT-PCR) and Western blotting (WB) Changes in the expression. Results: Triptolide 1.2 mg.L-1 inhibited the proliferation of C6 glioma cells in a dose-dependent manner. Triptolide treated C6 glioma cells for 24 h and then treated with 1.2 mg .The effect of L-1 on apoptosis was most obvious. Triptolide down-regulated the expression of Bcl-2 mRNA and protein in a dose-dependent manner, while the expression of Bax mRNA and protein increased . Conclusion: Triptolide has good growth inhibition and apoptosis induction on C6 glioma cells, and its mechanism may be related to the expression of Bcl-2 and Bax mRNA and protein.