摘要
实验运用PC12细胞系研究6-羟多巴胺的细胞毒性作用以及α-硫辛酸抗6-羟多巴胺毒性的作用及其机制。用MTT法测定显示6-OHDA使细胞存活率降低至56.8%,细胞突起变短、胞质浓缩、核质深染,细胞贴壁能力下降,胞膜损伤。原位末端dUTP标记法(TUNEL)显示阳性标记细胞,表明6-OHDA引起PC12细胞产生坏死和凋亡。流式细胞仪分析表明6-OHDA作用后凋亡细胞比例达20.09%。运用α-硫辛酸预处理后,能明显预防6-OHDA的毒性作用,可使细胞存活率上升,凋亡细胞比例降低至3.09%,α-硫辛酸的作用与提高细胞内超氧化物歧化酶(SOD)活力和还原型谷胱甘肽(GSH)含量有关。
PC12 cell line, a clonal cell line derived from a pheochromocytoma of rat adrenal medulla, was used as a model of dopaminergic neuron in vitro to study the effect of α-lipoic acid on the 6-OHDA induced apoptosis. The results from MTT method show that 6-OHDA decreased the cell survival rate obviously. Through TUNEL (TdT-mediated dUTP-biotion nick end labeling) and Flow cytometer (FCM) detection, we found that 6-OHDA triggered cell apoptosis and induced necrosis. It was confirmed by the different percentage of cell survival rate and apoptosis concluded from FCM and MTT. α-lipoic acid was used as antioxidant to protect the cell from 6-
OHDA's injury . The result indicateed that α-lipoic acid can partly prevent apoptosis induced by 6-OHDA but fail to prevent necrosis since it can decrease the apoptotic cell from 20.09 % to 3.09 % , just as increased cell survival rate from 56.8 % to 72.6 % but can not reach the normal level showed by MTT assay. Biochemical approach showed the cell's antioxidant ability especial for SOD activity and GSH content increased after the treatment of α-lipoic acid. The data suggest that α-lipoic acid may protect PC12 cells from apoptosis induced by 6-OHDA through the antioxidant path.
出处
《实验生物学报》
CSCD
北大核心
2001年第1期65-70,共6页
Acta Biologiae Experimentalis Sinica
