摘要
目的研究类叶升麻苷抗鱼藤酮致多巴胺能神经元SH-SY5Y细胞损伤,对帕金森病相关蛋白Parkin、α-突触核蛋白(α-Synuclein,α-Syn)表达的影响,探讨类叶升麻苷抗细胞损伤的分子机制。方法化学比色法测定细胞培养液乳酸脱氢酶(lactate dehydrogenase,LDH)活性,蛋白质印迹法(Western blot)检测Parkin、α-Syn的表达,免疫荧光法检测SH-SY5Y细胞α-Syn分布。结果①类叶升麻苷(10,20或40mg.L-1)可明显降低鱼藤酮诱导的SH-SY5Y细胞乳酸脱氢酶(LDH)的释放;②0.5μmol.L-1的鱼藤酮处理SH-SY5Y细胞48h能引起Parkin蛋白的明显降解及α-Syn蛋白二聚体增加;③预先用类叶升麻苷(10,20或40mg.L-1)处理细胞,能够有效减少鱼藤酮诱导的Parkin蛋白的降解,呈浓度依赖性;并能够抑制鱼藤酮诱导的α-Syn蛋白二聚体增加及α-Syn阳性细胞数增加。结论类叶升麻苷对鱼藤酮致SH-SY5Y细胞损伤具有神经保护作用,其作用机制可能与减少Parkin蛋白的降解和抑制α-Syn蛋白的二聚体形成有关。
Aim To investigate the neuroprotective effect of acteoside against rotenone-induced cell damage in SH-SYSY cells and the effect of acteoside on the expression of Parkinson disease （PD） -related proteins Parkin and α-Synuclein （α-Syn）, and to discover the underlying molecular mechanism of neuroprotection by acteoside. Methods The activity of lactate dehydrogenase（LDH） was measured by spectroscopy. Expressions of Parkin and α-Syn were studied using Western blot analysis, and the distribution of α-Syn was analyzed using immunofluorescence technique. Results （1） Acteoside （10, 20 or 40 mg· L^-1） pretreatment for 6 h markedly reduced the release of LDH induced by 0. 5 μmol· L^-1 rotenone; （2） Treatment with 0. 5 μmol· L^-1 rotenone for 48 h led to significant Parkin cleavage and increased formation of α-Syn protein dimer; （3） Pretreatment of SH-SYSY cells with acteoside （ 10, 20 or 40 mg·L^-1 ） for 6 h markedly reduced the cleavage of Parkin induced by 0.5 μmol· L^-1 rotenone in a concentration-dependent manner, inhibited the aggregation of α-Syn and decreased α-Syn-positive SH-SYSY cells. Conclusion These findings suggest that pretreatment of acteoside has a potent neuroprotective effect against rotenone-induced SH-SYSY cells damage and its mechanism might involve in reducing the cleavage of Parkin and inhibitng α-Syn expression induced by rotenone.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2007年第2期161-165,共5页
Chinese Pharmacological Bulletin
基金
国家重点基础研究发展计划(973计划)项目子课题(No2004CB518902)
国家自然科学基金项目资助课题(No30472164)
