论文部分内容阅读
目的:观察中药单体姜黄素对ActD/TNF-α协同诱导PC12细胞凋亡的影响,并探讨其机制。方法:采用MTT法确定实验药物的最佳浓度;Hoechst33258荧光染色法观察PC12细胞的凋亡;JC-1荧光分子探针检测线粒体膜电位;Real Time PCR检测凋亡基因Bcl-2/Bax的表达。结果:ActD/TNF-α协同作用可导致PC12细胞的活力降低(P<0.05);出现核固缩、核碎裂现象的细胞增多,细胞凋亡率增高(P<0.05);细胞线粒体膜电位下降;细胞内抗凋亡基因Bcl-2的表达降低(P<0.05)。经姜黄素(5μmol/L)处理后,PC12细胞的活力增强(P<0.05);细胞核固缩、核碎裂现象减少,细胞凋亡率下降(P<0.05);细胞线粒体膜电位上升;细胞内抗凋亡基因Bcl-2的表达增强(P<0.05)。结论:姜黄素可拮抗ActD/TNF-α引起的PC12细胞凋亡,可能与升高线粒体膜电位,促进抗凋亡基因Bcl-2的表达有关。
OBJECTIVE: To observe the effect of curcumin, a traditional Chinese medicine, on the apoptosis of PC12 cells induced by ActD / TNF-α and its mechanism. Methods: MTT method was used to determine the optimal concentration of the experimental drugs. Hoechst33258 fluorescence staining was used to observe the apoptosis of PC12 cells. The mitochondrial membrane potential was detected by JC-1 fluorescent probe. The expression of Bcl-2 / Bax protein was detected by Real Time PCR . Results: The synergistic effect of ActD / TNF-α resulted in a decrease in the viability of PC12 cells (P <0.05). The number of cells with nuclear condensation and nuclear fragmentation increased and the apoptosis rate increased (P <0.05). The mitochondrial membrane potential (P <0.05). The expression of Bcl-2 in the cells was decreased. After treated with curcumin (5μmol / L), the viability of PC12 cells was enhanced (P <0.05). The number of nuclear condensation and fragmentation was decreased and the apoptosis rate was decreased (P <0.05). The mitochondrial membrane potential The expression of anti-apoptotic gene Bcl-2 was enhanced (P <0.05). CONCLUSION: Curcumin can antagonize ActD / TNF-α-induced PC12 cell apoptosis, which may be related to increasing mitochondrial membrane potential and promoting the expression of anti-apoptotic gene Bcl-2.