安颤灵对泛素-蛋白酶体通路损伤细胞模型的保护作用

目的 利用中药血清药理学方法探讨安颤灵含药血清对lactacystin诱导PC12细胞损伤的保护作用及其可能机制. 方法 按照血清药理学实验方法制备安颤灵含药血清,将含药血清分为大、中、小剂量组(分别按7.3 g/kg、3.65 g/kg、1.83 g/kg添加药物),用lactacystin建立PC12细胞泛素-蛋白酶体系统(UPS)损伤模型.MTT方法检测各组的细胞活力.将对数生长期的PC12细胞分为正常组(DMEM+10%空白鼠血清)、损伤组(DMEM+10%空白鼠血清+10 μmol/Llactacystin)、安颤灵中剂量组(含DMEM+10%中剂量安颤灵含药血清+10 μmol/L lactacystin),通过免疫组化方法,观察各组细胞α-突触核蛋白、酪氨酸羟化酶(TH)表达的情况. 结果 与大、小剂量组相比,中剂量组细胞的存活率明显升高,差异有统计学意义(P＜0.05);与正常组相比,损伤组细胞α-突触核蛋白阳性细胞数增多,而TH阳性细胞明显减少,差异有统计学意义(P＜0.05);与损伤组相比,安颤灵中剂量组细胞α-突触核蛋白阳性细胞数减少,TH阳性细胞明显增加,差异有统计学意义(P＜0.05). 结论 安颤灵含药血清能改善细胞UPS的功能,降低细胞内α-突触核蛋白的聚集并上调TH表达,从而起到提高细胞存活率,保护神经元的作用.

Protective effect of Anchanling Oil lactacystin-induced damage of the ubiquitin-proteasome pathway in PCI2 cells

Objective To explore the protective effect of Anchanling serum on PCI2 cell injury induced by lactacystin and the possible mechanism.Methods The sem were obmined from rats with intragastric administration of Anchanling at small,medium and high doses.Lactacystin was used to induce proteasomal dysfunction in PCI2 cells,and the cell viability was evaluated with MTT assay.The PC 12 cells in exponential growth were divided into normal control,lactacystin injury and Anchanling-treated groups with corresponding treatments,and theexpressions of alpha-synuclein and tyrosine hydroxylase(TH)were assayed using immunohistochemical staining.Results The serum of rats with medium-dose Anchanling treatment group resulted in the highest viability of the PC 12 cells among the 3 dose groups(P＜0.05).Compared with the normal control group,lactacystin exposure significantly increased the number of alpha-synuclein-positive cells but decreased the TH-positive cells (P＜0.05);treatments with the drug-containing sera significantly lowered alpha-synuclein-positive cells and increased TH-positive cells in the PC 12 cells with lactacystin exposure(P＜0.05).Conclusion Anchanling can improve the function of the ubiquitin-proteasome system,reduce the intracellular aggregation of alpha-synuclein,and upregulate TH expression in the PC 12 cells to promote the cell survival and offer neuronal protection effects.