桦褐孔菌醇对人宫颈癌HeLa细胞凋亡的影响

目的：研究桦褐孔菌醇（Inotodi01）体外诱导人宫颈癌HeLa细胞凋亡的作用，并探讨其可能的作用机制。方法：以不同浓度的Inotodiol处理HeLa细胞24h、48h和72h，采用MTT法检测细胞增殖情况，透射电镜观察细胞超微结构变化，TUNEL法检测细胞凋亡，免疫细胞化学法检测caspase-3蛋白的表达。结果：Inotodiol能抑制HeLa细胞的增殖，且呈时间-剂量依赖关系（P〈0．05，P〈0．01）；超微结构观察可见细胞体积缩小、核固缩、核内异染色质增多并边集、凋亡小体形成等典型的凋亡形态学改变；TUNEL法显示，经50μg/ml的lnotodiol处理48h，HeLa细胞发生凋亡；免疫细胞化学结果表明，与对照组相比，Inotodiol组HeLa细胞中caspase-3蛋白表达上调（P〈0．01）。结论：在一定浓度范围内，Inotodiol可在体外抑制HeLa细胞增殖并诱导其凋亡，其作用机制可能与其上调easpase-3蛋白的表达有关。

Effects of Inotodiol on Apoptosis of HeLa Cells in Human Cervical Cancer

Objective： To investigate the effect of Inotodiol on HeLa cells in vitro, and to explore its mechanism. Methods： The HeLa cells were treated with Inotodiol at different concentrations for 24 h, 48 h and 72 h, and the inhibitory rate of cell proliferation was assessed detected by melhyl thiazolyl tetrazolium （MTT） assay. Uhrastructure changes of HeLa cells were examined by electronmicroscope. The TUNEL assay was used to detect the apoptosis of HeLa ceils. The expression of Caspase-3 protein was analyzed by immunocellulerchemical method. Results： Inotodiol can inhibit the proliferation of HeLa ceils, and in a dose-time dependent manner （P〈0.05, P〈0.01）; the cells were smaller with karyopycnosis, heterochromatin increased and edge set. Apoptosis body can also be observed as typical changes of apoptosis under transmission electron microscope. TUNEL assay indicated that the apoptosis of HeLa ceils treated with 50μg/ml Inotodiol for 48 h was induced. The immunocytochemistry staining showed that the expression levels of caspase-3 proteins of HeLa cells treated with Inotodiol were up-regulated compared with control groups （ P 〈 0.01 ）. Conclusion ： In a certain concentration range, Inotodiol can inhibit the proliferation of HeLa cells and induce the apoptosis in vitro. This effect may be associated with the up-regulated expressions of caspase-3 proteins.