27-羟基胆固醇在神经细胞和星形胶质细胞共培养中的毒性作用  

The cytotoxicity of 27-hydroxycholesterol on co-cultured SH-SY5Y cells and C6 cells

作　　者：王慧[1] 肖荣[1] 麻微微[1] 席元第[1] 卢言慧[1] 荣红国 安宇[1] 

机构地区：[1]首都医科大学公共卫生学院环境毒理学北京市重点实验室,北京100069

出　　处：《毒理学杂志》2017年第1期1-5,共5页Journal of Toxicology

基　　金：国家自然科学基金(81330065)

摘　　要：目的探讨在神经细胞和星形胶质细胞共培养条件下,27-羟基胆固醇(27-OHC)的神经毒性作用。方法先将神经细胞(SH-SY5Y细胞系)和星形胶质细胞(C6细胞系)分别单独培养,经不同剂量的27-OHC处理不同时间后,用CCK-8实验检测细胞活力;继而采用SH-SY5Y细胞和C6细胞共同培养,实验分为4组:对照组、27-OHC低剂量组(5μmol/L组)、27-OHC中剂量组(10μmol/L组)和27-OHC高剂量组(20μmol/L组);用光学显微镜观察细胞形态改变,用化学比色法检测乳酸脱氢酶(LDH)活性水平,用流式细胞仪对SH-SY5Y细胞和C6细胞的线粒体膜电位(MMP)进行检测。结果随着27-OHC浓度升高,各组细胞数量逐渐下降,细胞出现萎缩、形态不规则;LDH活性逐渐升高(P<0.05),细胞线粒体膜电位降低(P<0.05)。结论 27-OHC引起神经细胞和星形胶质细胞的形态发生改变、细胞数量减少、细胞膜破损及线粒体膜电位降低,且这种毒性作用显示出一定的剂量反应关系。Objective To investigate the cytotoxicity of the 27-hydroxycholesterol（ 27-OHC） on co-cultured nerve cell and astrocyte. Methods Firstly,neurocytes（ SH-SY5Y cells） and astrocytes（C6 cell lines） were cultured respectively,cell viability was tested by CCK-8 after different doses of 27-OHC incubation at different times. Then SH-SY5Y cells and C6 cells were co-cultured. Four groups were the following： control group,Low dose group of 27-OHC treated（ 5 μmol/L group）,middle dose group of 27-OHC treated（ 10 μmol/L group） and high dose group of 27-OHC treated（ 20 μmol/L group）. Cell morphology was observed by optical microscope. The level of lactate dehydrogenase（ LDH） activity was detected by chemical assay. The mitochondrial membrane potential（ MMP） of SH-SY5Y cells and C6 cells were detected by flow cytometry. Results The cells in 27-OHC treated groups had a lower cell viability than the control group,also the number was decreasing and the cells had irregular shape. Compared with control group,27-OHC treated groups had higher LDH activity（ P〈0. 05） and lower mitochondrial membrane potential（ P〈0. 05）. Conclusion 27-OHC might cause cell morphological changes,reduce the number of cell,damage cell membrane structure and decrease mitochondrial membrane potential. Moreover,the toxicity showed a dose-response relationship.

关 键 词：27-羟基胆固醇 神经细胞 星形胶质细胞 共培养 神经毒性 

分 类 号：R99[医药卫生—毒理学]