【摘 要】
：
目的 观察干预细胞核因子-κB(NF-κB)信号通路对星形胶质细胞(AST)自噬、凋亡和坏死的影响. 方法 取纯化AST分成3组:(1)空白组:加入2μL PBS液1h后,再加入2μLPBS液;(2)激活
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目的 观察干预细胞核因子-κB(NF-κB)信号通路对星形胶质细胞(AST)自噬、凋亡和坏死的影响. 方法 取纯化AST分成3组:(1)空白组:加入2μL PBS液1h后,再加入2μLPBS液;(2)激活组:加入2μL PBS液1h后,再加入2μL NF-κB激活剂PMA(终浓度1μmol/L);(3)抑制组:先加入2 μL NF-κB阻断剂PDTC(终浓度20 μmol/L)预处理lh,再加入2μL PMA(终浓度1μmol/L).各组细胞分别处理1、6、12、24、48 h,应用免疫细胞化学荧光法测定NF-κB蛋白的表达,应用AnnexinV/PI染色法检测凋亡和坏死,应用Western blotting检测自噬蛋白Beclin 1表达情况. 结果 (1)随着作用时间增加,NF-κB蛋白表达量也逐渐增高,24 h时空白组、激活组及抑制组表达量分别为0.27±0.46、3.13±0.35、2.00±0.53,差异有统计学意义(P＜0.05).(2)各组细胞凋亡及坏死程度也随着作用时间增加而相应增高.激活组AST凋亡程度总体高于空白组与抑制组,12、24、48 h时间点3组细胞两两比较差异均有统计学意义(P＜0.05).激活组AST坏死程度总体高于空白组与抑制组,24 h、48 h时间点3组细胞两两比较差异均有统计学意义(P＜0.05).(3)激活组Beclin 1蛋白各时间点均较抑制组及空白组明显增加,6h时空白组、激活组及抑制组Beclin 1蛋白表达量分别为0.17±0.01、0.59±0.03、0.51 ±0.03,两两比较差异均有统计学意义(P＜0.05). 结论 PMA诱发的NF-κB活化能够引起AST不同程度的凋亡、坏死及自噬进程,早期以凋亡和自噬为主,晚期以坏死为主,而PDTC能够在一定程度上抑制NF-κB通路所诱导的这些死亡进程.
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