摘要
目的 :研究bcl 2家族 (主要是bcl xl和bax)基因及相关蛋白是否参与水溶性淀粉样蛋白对培养的大鼠嗜铬细胞瘤细胞的毒性作用。方法 :应用电镜和DNA电泳判断细胞损伤途径 ,应用RT PCR ,免疫细胞化学和免疫印迹判断bcl 2家族及其相关蛋白是否参与淀粉样蛋白的损伤作用。结果 :10 μmol/LAβ2 5 35作用 2 4h后 ,电镜和DNA电泳均显示Aβ2 5 35可以导致PC12细胞凋亡 ;RT PCR表明药物作用后 6h ,bcl xlmRNA表达量减少而baxmRNA表达量增加 ;免疫印迹和免疫细胞化学结果显示药物作用后 2 4h ,Bax蛋白增加而Bcl xl无明显改变。结论 :水溶性淀粉样蛋白可以导致神经细胞凋亡 ,bcl 2家族参与了作用环节 ,对阿尔茨海默病的发病起着重要的作用。
Objective:To investigate whether bcl 2 family genes (mainly bcl xl and bax) were involved in Aβ induced differentiated PC12 cells death. Methods:The electron microscope and DNA electrophoresis were used to clarify the morphological change of Aβ induced PC12 cell death, and RT PCR, immunocytochemistry and Western blot were applied to investigate if bcl 2 family members were involved. Results:After treatment by Aβ 25 35 (10 μmol/L) for 24 h, morphological evidences for apoptosis nuclear condensation and DNA ladder were observed in PC12 cells. RT PCR, immunocytochemistry and western blot showed that down regulation of bcl xl mRNA appeared after 6 h of the treatment while there was no change in Bcl xl protein expression after 24 h. On the other hand, bax mRNA increased slightly and Bax protein expression increased significanly. Conclusion: These findings demonstrated that bcl xl and bax are the main effectors of Aβ and may participate in neuronal degeneration in Alzheimer's disease.
基金
上海市自然科学基金 ( 99ZB14 0 19)资助项目
