鱼藤酮对PC12细胞Caspase-3,Fas和Fasl的作用及与凋亡的关系  

Effect of rotenone on Caspase-3, Fas and Fasl and relationship with apoptosis in PC12 cells

作　　者：赵黔鲁[1] 柴锡庆[1] 

机构地区：[1]河北医科大学研究生学院,河北石家庄050017

出　　处：《第四军医大学学报》2005年第20期1888-1891,共4页Journal of the Fourth Military Medical University

摘　　要：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.[Abstract] AIM： To investigate the effect of rotenone on Caspase-3, Fas and Fasl and relationship with apoptosis in PC12 cells. METHODS： PC12 cells were treated with different concentrations of rotenone and the effect of rotenone on the ratio of apoptosis was tested by flow cytometry, 1. 0 μmol/L rotenone was added into PC12 cells culture medium at different times and the protein expression of Caspase-3 and Fas/Fasl was also tested using FCM. The morphological changes of apoptosis were observed under transmitting electron microscopy （TEM）. RESULTS： The ratio of apoptosis in control group was 0. 0199 ± 0. 0020. In the trial groups treated with different doses of rotenone for 2 d, the ratio increased as 0.0305 ± 0.0030, 0.0486 ± 0. 0073 , 0. 0313 ±0. 0041 and 0. 0285 ± 0. 0031, respectively, significantly different from that in control group （ P 〈 0.02）. The 1. 0 μmol/L rotenone had marked effect （ P〈0.01 ）. When rotenone concentrations exceeded 1. 0 μmol/L, the effects decreased in a dose-dependent manner. FI of Caspase-3, Fas and Fasl in control group was 1. 0 ± 0.03, 1. 0 ± 0.02 and 1. 0 ± 0. 04, respectively. After treatment with 1. 0 μmol/L rotenone for 3, 6, 12, 24 and 48 h, the expression of Caspase-3, Fas and Fasl was observed in each group. The expression of Fas/Fasl was most significant after rote- none treatment for 6 h （ P 〈 0.01 ） and the expression of Caspase-3 was most significant after rotenone treatment for 12 h （ P 〈 0.01 ）. The morphological changes under TEM were shown as unsmooth membrane, decreased cellular protruding, small ratio of nucleus/cytoplasm, increased space in cytoplasm, increased gap of periphery in nucleus uncertainly, and increased poikilochromatin collected in edge. CONCLUSION： Rotenone may damage the PC12 cells by activation of Caspase-3 and Fas/ Fasl apoptosis pathways.

关 键 词：鱼藤酮 细胞凋亡 帕金森病 显微镜检查 电子 

分 类 号：R363.13[医药卫生—病理学] R742.5[医药卫生—基础医学]