D2/D3受体激动药罗匹尼罗对抗鱼藤酮介导多巴胺能细胞凋亡的研究

目的:探讨D2/D3多巴胺受体激动药罗匹尼罗对多巴胺能细胞是否具有抗凋亡作用并研究其机制。方法:使用多巴胺能细胞株SH-SY5Y,以高剂量呼吸链抑制药鱼藤酮诱导细胞凋亡建立细胞凋亡模型。通过Hoechst染色观察并计数凋亡的细胞,应用免疫印迹(Westernblot)方法检测促凋亡基因caspase 9, cleaved caspase 9,cleaved caspase 3,PARP,cleaved PARP的表达。结果:高剂量鱼藤酮处理细胞24h后其凋亡比例明显升高,罗匹尼罗(10~(-5),10~(-4)mol·L~(-1))预处理2h能显著减少细胞凋亡的比例。免疫印迹方法显示10~(-4)mol·L~(-1)罗匹尼罗预处理2h能减少caspase 9,caspase 3,PARP的剪切。结论:罗匹尼罗能有效抑制鱼藤酮诱导细胞凋亡,其机制可能是通过抑制内源性通路中caspase 9剪切而实现的。

Protective effects of D2/D3 receptor agonist,ropinirole,against rotenone induced apoptosis of dopaminergic cells

AIM:To observe the protective effects of D2/D3 receptor agonist ropinirole against rotenone induced apoptosis of dopaminergic cells and investigate the mechanisms.METHODS:Rotenone,high dose respiratory inhibitor,was used to establish apoptotic models in dopaminergic cell line SH-SY5Y.Apoptotic cells were calculated after Hoechst staining.Apoptotic-related genes such as caspase 9, cleaved caspase 9,cleaved caspase 3,PARP and cleaved PARP were detected by the method of Westernblot.RESULTS:High percentage apoptotic cells were observed after treated with high doses rotenone.Ropinirole at dose(10~(-5),10~(-4)mol·L~(-1)) pretreated for 2 hours could significantly inhibit cell apoptosis.The expression of cleaved caspase 9,cleaved caspase 3 and cleaved PARP detected by Westemblot were decreased after pretreatment with 10~(-4)mol·L~(-1) ropinirole.CONCLUSION:D2/D3 receptor agonist ropinirole has anti-apoptotic effects in SH-SYSY dopaminergic cell line through the inhibition of caspase 9,caspase 3 and PARP cleaves.