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目的:探讨鱼藤酮对PC12细胞Caspase3,Fas和Fasl的作用及与凋亡的关系.方法:应用流式细胞仪(flowcytometry,FCM)检测不同浓度鱼藤酮对体外培养大鼠肾上腺嗜铬细胞瘤(PC12)细胞株凋亡率的影响,并检测1.0μmol/L鱼藤酮干预不同时段对Caspase3、Fas/Fasl蛋白表达的影响,透射电镜观察凋亡形态学的改变.结果:对照组凋亡率为0.0199±0.0020,经0.1,1.0,2.0和3.0μmol/L不同浓度鱼藤酮干预2d,凋亡率明显增加其变换值分别为:0.0305±0.0030,0.0486±0.0073,0.0313±0.0041和0.0285±0.0031,与对照组比较有显著差别(P<0.02),实验组中以1.0μmol/L作用最显著(P<0.01),当鱼藤酮浓度大于1.0μmol/L时,其作用呈剂量依赖性递减.对照组Caspase3,Fas和FaslFI值分别为1.0±0.03,1.0±0.02和1.0±0.04;经鱼藤酮干预3,6,12,24和48h,各组均有Caspase3,Fas和Fasl表达,其中以鱼藤酮干预6h时Fas/Fasl表达最明显(P<0.01),而12h时Caspase3表达最明显(P<0.01).电镜下观察到细胞膜不光滑,细胞突起减少,核质比小,细胞质空间增大,核周隙增大、不规则,异染色质增多、边集.结论:鱼藤酮可通过激活Caspase3,Fas/Fasl凋亡途径损伤PC12细胞.
OBJECTIVE: To investigate the effect of rotenone on the expression of Caspase3, Fas and Fasl in PC12 cells and its relationship with apoptosis.Methods: Flow cytometry (FCM) was used to detect the effect of rotenone on PC12 expression in rat adrenal pheochromocytoma (PC12) The effect of 1.0 μmol / L rotenone on the expression of Caspase3 and Fas / Fasl protein was observed at different time points and the morphological changes of apoptotic cells were observed by transmission electron microscope.Results: The apoptosis rate in the control group was 0.0199 ± 0.0020 , The apoptosis rate was significantly increased after treated with different concentrations of 0.1, 1.0, 2.0 and 3.0μmol / L rotenone for 2 days. The conversion values were 0.0305 ± 0.0030,0.0486 ± 0.0073,0.0313 ± 0.0041 and 0.0285 ± 0.0031, respectively, compared with the control group (P <0.01). The effect of 1.0 μmol / L was the most significant (P <0.01) in the experimental group, while the dose-dependent decrease was observed when the rotenone concentration was greater than 1.0 μmol / L. In the control group, the expressions of Caspase3, Fas and FaslFI The values were 1.0 ± 0.03, 1.0 ± 0.02 and 1.0 ± 0.04, respectively. The expressions of Caspase3, Fas and Fasl in each group were significantly different at 3, 6, 12, 24 and 48 h after intervention with rotenone (P <0.01), while the expression of Caspase3 was the most obvious at 12h (P <0.01) The results showed that rotenone could activate caspase3 and Fas / Fasl pathway by inducing caspase3 and Fas / Fas1 pathway, and the cell membrane was not smooth, the number of protuberances was decreased, the ratio of nucleus to cytoplasm was small, the cytoplasm space was increased, the perinuclear space was enlarged, irregular and heterochromatin was increased. Damage PC12 cells.