灵芝多糖对Aβ_(25-35)诱导的PC12细胞损伤的保护性研究

Protective effect of ganoderma lucidum polysaccharide against-amyloid-induced neurotoxicity in PC12 cells

目的观察灵芝多糖(GLP)对β-淀粉样蛋白(Aβ25-35)诱导的PC12细胞损伤的保护作用。方法将Aβ25-35或(和)不同浓度的GLP加入体外培养的PC12细胞中,用MTT检测PC12细胞活力的变化;以DCFH-DA来检测细胞内活性氧(ROS)水平的改变;通过Western Blotting来检测Bcl-2和Bax的表达水平。结果 Aβ25-35处理36h后的PC12细胞存活率仅为对照的60.7%,细胞内ROS水平上升到原来的222.7%,同时Bcl-2/Bax比值下降,为对照组的60.0%。经不同浓度的GLP预处理后,细胞存活率均显著提高,分别能达到69.7%,80.7%和88.3%(P<0.01);10g/ml GLP预处理PC12细胞24h后,细胞内ROS水平下降至正常组的160.6%,Bcl-2/Bax比值升高到93.6%,P值均<0.01。结论 GLP对Aβ25-35诱导的PC12细胞损伤有保护作用。

Abstract： Objective To observe the protection effects of ganoderma lucidum polysaccharide（GLP） on-amyloid（A） induced neurotoxieity in PC12 cells. Methods Neurotoxicity was induced by addition of Aβ25-35 into cultures of PC12 cells and different doses of GLPS were administrated 2h before addition of Aβ25-35 into PC12 cell cultures. Cell viability, ROS level and the expression of Bcl-2 and Bax were measured using MTF, DCFH-DA and western blotting, respectively. Results The percentage of viability of PC12 cells treated with AI325-35 for 36h was 60.7% of contrD1 group. In addition, treatment with Aβ25_35 increased cellular ROS level （222.7% of the control value）and reduced the ratio of Bel-2/Bax （60.0% of the control value）. Pretreatment with single dose of GLP at 2,10, and 50μg/ml increased cell viability by 65% ,80.7% ,and 88.3% （P 〈 0.01 ）, respectively. In parallel, treatment with GLP at 10 μg/ml ignificantly decreased the level of intracellular ROS（ 160.6% of the control value）and moderated the ratio of Bcl-2/Bax （93.6% of the control value） ,all P value 〈0.01. Conclusion The results suggest that pretreatment of GLP protects PC12 cells against Aβ25-35 induced neurotoxicity.