蛋白酶体抑制剂诱导的帕金森病模型中galectin-1含量变化的研究

The study on the changes of galectin-1 in the parkinson's cell model induced by proteasomal inhibition

目的检测蛋白酶体抑制剂PSI诱导的帕金森病(PD)细胞模型中,β半乳糖苷结合血凝素(galectin-1)的含量变化及其在PC12细胞内的分布。方法PSI作用于PC12细胞后,荧光染料染色检测细胞凋亡率;HE染色观察细胞形态学变化。对PSI诱导的PD细胞模型进行蛋白质组学研究,应用免疫细胞化学的方法研究galectin-1在PC12细胞内分布,应用免疫印迹的方法研究此模型中galectin-1的含量变化。结果10μMPSI作用于PC12细胞24小时,细胞凋亡比例增加(P<0.05),胞浆内出现了嗜酸性包涵体。蛋白质组学和免疫印迹研究均提示galectin-1的含量在细胞模型组明显减少(P<0.05)。galectin-1主要分布于PC12细胞的胞膜、胞浆及细胞核内。结论蛋白酶体抑制剂PSI作用于PC12细胞,模拟了人类PD,在此模型中galectin-1减少,提示此蛋白可能在PD的发病机制中起作用。

Objective To explore the distribution of galectin-1 in PC12 cells and its expression change in the Parkinson＇s cell model induced by proteasomal inhibitor PSI.Methods Fluorescent DNA stain was used to evaluate the apoptotic ratio of PC12 cells treated with PSI.The morphology of PC12 cells treated with PSI was observed after HE staining.A proteomic approach was used to determine proteins in cells that were differentially expressed following treatment of PSI.The distribution of galectin-1 in PC12 cells was studied with the immunocytochemistry.The expression change of galectin-1 was studied with Western blot technique.Results According to fluorescent DNA stain observation,10μM PSI administration for 24 hours resulted in cell apoptosis（P0.05）.Eosinophilic cytoplasmic inclusions were detected in PC12 cells treated with PSI after HE staining.The proteomic study and Western blot showed the expression of galectin-1 decreased（P0.05）.Immunohistochemical study showed that galectin-1 was distributed on the cell membrane,in the cytoplasm and nucleus.Conclusion The model of Parkinson＇s disease could be induced in PC12 cells by treatment of proteasomal inhibitor PSI.In this model galectin-1 decreased,indicating it may be an important factor in the pathogenetic mechanism of this disease.