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目的:探讨虫草素对鱼藤酮诱导的PC12细胞凋亡是否具有保护作用。方法:以1μmol·L~(-1)鱼藤酮制备帕金森病(Parkinson’s disease,PD)体外细胞模型,用MTT法测定细胞存活率;比色法测定LDH释放率;Hoechst 33258核染色法观察不同时间细胞凋亡形态;DNA Ladder检测细胞凋亡;流式细胞仪分析细胞凋亡比率。结果:1不同浓度鱼藤酮作用不同时间均能明显降低PC12细胞存活率(P<0.001),1μmol·L~(-1)时效果最为显著。2低浓度虫草素(10-6~10-2μmol·L~(-1))除24 h促进正常PC12细胞增殖外其余均无明显影响;但1~102μmol·L~(-1)虫草素能够抑制PC12细胞增殖(P<0.05,0.01,0.001)。3将0.4×10-6,2.0×10-6,8.0×10-6μmol·L~(-1)虫草素分别与鱼藤酮同时给药,作用24,48和72 h后均能使细胞存活率显著上升(P<0.05);LDH释放率显著降低(P<0.05);Hoechst 33258核染色中细胞核出现致密蓝色亮点的细胞明显减少;虫草素干预48 h后,DNA片段化现象减少或消失,经流式细胞仪检测,细胞总凋亡率明显降低(P<0.001)。结论:虫草素能抑制鱼藤酮诱导的PC12细胞凋亡,具有潜在的保护作用。
Objective: To investigate whether cordycepin can protect rotenone-induced PC12 cells from apoptosis. Methods: Cell model of Parkinson’s disease (PD) was established by using 1 μmol·L -1 rotenone. The cell viability was measured by MTT assay. The release rate of LDH was determined by colorimetric assay. Hoechst 33258 nuclear staining was used to observe cell viability at different times Cell apoptosis morphology; DNA Ladder detection of apoptosis; flow cytometry analysis of apoptosis rate. Results: 1 Different concentrations of rotenone can significantly reduce the survival rate of PC12 cells (P <0.001), and the effect of 1μmol·L -1 was the most significant. 2 Cordyceps sinensis (10-6 ~ 10-2μmol·L -1) had no significant effect on the proliferation of normal PC12 cells except 24 h, but cordycepin 1 ~ 102μmol·L -1 could Inhibit PC12 cell proliferation (P <0.05, 0.01, 0.001). 3 0.4 × 10-6,2.0 × 10-6,8.0 × 10-6μmol·L -1 Cordycepin were administered with rotenone at the same time, after 24, 48 and 72 h, the cell survival rate was significantly (P <0.05). The release rate of LDH was significantly decreased (P <0.05). The number of cells with dense blue spots in nucleus of Hoechst 33258 nucleus decreased obviously. The DNA fragmentation decreased or disappeared after 48 h intervention of cordycepin. Flow cytometry showed that the total apoptosis rate was significantly decreased (P <0.001). Conclusion: Cordycepin can inhibit the apoptosis of PC12 cells induced by rotenone and has a potential protective effect.