灵芝孢子多糖LPS3对Lactacystin诱导PC12细胞损伤的影响

目的探讨灵芝孢子多糖LPS3对蛋白酶体抑制剂Lactacystin诱导PC12细胞损伤的影响。方法体外培养PC12细胞,建立Lactacystin诱导PC12细胞损伤的模型,用CCK-8法检测细胞活力,Annexin V-FITC/PI流式细胞法检测细胞调亡,免疫荧光法检测胞浆中α-synuclein聚集物的生成。结果模型组经20μmol·L-1Lactacystin处理24 h后,PC12细胞活力比空白对照组降低,仅为64.7%;经不同浓度的灵芝孢子多糖LPS3预处理后,细胞活性明显提高,且其细胞活性随多糖浓度升高而升高;Lactacystin可以诱导PC12细胞凋亡,处理24 h后细胞凋亡率为45.40%,经灵芝孢子多糖LPS3预处理后,细胞凋亡率明显降低;PC12细胞经Lactacystin处理后,胞浆内可见明显的颗粒状、小团块状α-synuclein聚集体的生成;而经灵芝孢子多糖LPS3预处理后,胞浆中α-synuclein的聚集体明显减少。结论灵芝孢子多糖LPS3对Lactacystin诱导的PC12细胞损伤有一定的保护作用,其作用机制可能与抑制α-synuclein聚集体的生成有关。

Effects of Ganoderma spore polysaccharide LPS3 on Lactacystin induced PC12 cells damage

OBJECTIVE To investigate the effect of Ganoderma spore polysaccharide LPS3 on PC12 cells treated with Lactacystin.METHODS The toxicity model was established by treating PC12 cells with Lactacystin in vitro.Cell viability was assayed with CCk-8 method.The aggregates of α-synuclein were determined by immunofluorescence cytochemistry staining.RESULTS After treated with Lactacystin(20 μmol·L~-1) for 24 h,the viability of PC12 cells depressed and the percentage viable cells was only 64.7%,when compared with control group.Pretreated with Ganoderma spore polysaccharide LPS3 at different concentrations,the cell viabilities were all increased significantly.The protective effect on cells increased as its concentration increased.Lactacystin induced apoptosis in PC12 cells,and the apoptotic rate was 45.40% after 24 h of treatment.Pretreated with Ganoderma spore polysaccharide LPS3,the cell apoptotic rate reduced.In addition,immunofluorescence cytochemistry staining showed that PC12 cells treated with Lactacystin had significantly fewer aggregates of α-synuclein in group pretreated with Ganoderma spore polysaccharide LPS3 than the unpretreated group.CONCLUSION Ganoderma spore polysaccharide LPS3 can protect PC12 cells from the injury induced by Lactacystin,and its mechanism may be associated with reducing the formation of aggregates of α-synuclein.