白藜芦醇通过Fas-线粒体双途径诱导U-87脑胶质瘤细胞凋亡

目的研究白藜芦醇（Res）对人脑胶质瘤细胞系U-87细胞增殖抑制和诱导凋亡作用,并探讨其分子机制。方法以人脑胶质瘤细胞系U-87细胞为靶细胞,应用四氮唑蓝（MTT）比色法测定细胞增殖活性;AnnexinV/碘化丙啶（PI）双标记和细胞形态学法检测U-87细胞凋亡;流式细胞仪（FCM）检测细胞Fas蛋白表达水平、线粒体跨膜电位（Δψm）、细胞色素C（Cytc）释放和Caspase-3活性变化。结果Res明显抑制U-87细胞的增殖,呈浓度及时间依赖性（P〈0.01）;20μmol/L和40μmol/LRes处理U-87细胞,AnnexinV/PI染色显示凋亡细胞明显增多,细胞凋亡率分别为42.57%和62%,同时细胞出现典型的凋亡形态改变;FCM检测显示Fas蛋白表达增高1.6～2.2倍,线粒体Δψm降低15%～63%,胞浆Cytc含量增加2.5～7.3倍,Caspase-3被激活,活性增高25%～112%。结论Res体外明显抑制U-87细胞的增殖,通过Fas-线粒体双途径诱导U-87细胞凋亡。

Resveratrol induced apoptosis of U-87 glioma cells through the Fas-mitochondria pathway

Objective To investigate the proliferation-inhibiting and apoptosis-inducing activity of resveratrol （Res） on human brain glioma c, ell line U-87 in vitro and the possible molecular mechanisms. Methods Human brain glioma cell line U-87 was used as the target cells to detect tile cell proliferating activity by a methyl thiazolyl tetrazolium （MTY） colorimetric assay. The apoptosis of U-87 cells was assessed with FITC-Annexin V and propidium iodide （PI） double staining and the mophological observation. Expression of Fas protein, mitochondrial inner membrane potential （△ψm）, cytoehrome c （Cyt c ）release, and caspase-3 aetivity were analyzed by flow cytometry （FCM）. Results Res significantly inhibited the proliferation of U-87 eells in a dose- and time-dependent manner （P 〈0.01 ）. After treated with 20 mol/L and 40 mol/L of Res, the apoptosis rate of U-87 cells deteeted Annexin V/PI staining obviously increased to 42.57% - 62%, and the typical apuptotie morphological changes of U-87 eells were observed. FCM detected the explession of Fas protein increased by 1.6 -2.2 times, aeeompanied with a decrease in mitoehondfia AtOm by 15% -63% and the increased release of Cyt e by 2.5 -7.3 times from mitochondria to cytoplasm. The activity of easpase-3 was significantly enhaneed by 25% - 112%. Conclusion Res greatly inhibits the proliferation of U-87 cells and induces the apoptosis of U-87 cells via the Fas-mitochondria pathway in vitro.