骨髓间充质干细胞对TAG1介导的PC12细胞凋亡的保护作用
Protective effect of bone marrow mesenchymal stem cells on PC12 cells apoptosis mediated by TAG1.
作者信息
Zhang Yu-Zhen, Lou Ji-Yu, Bai Hong-Ying, Wang Yun-Liang, Li Jin-Feng, Yin Hong-Lei
机构信息
Department of Neurology, The Second Affiliated Hospital of Zhengzhou University Zhengzhou, P. R. China.
Department of Neurology, 148 Hospital of PLA Zibo, P. R. China.
出版信息
Int J Clin Exp Pathol. 2015 Oct 1;8(10):12093-100. eCollection 2015.
OBJECTIVE
This study aims to explore the protection effect of bone marrow mesenchymal stem cells (BMSCs) on PC12 cells apoptosis mediated by transient axonal glycoprotein 1 (TAG1).
METHODS
PC12 cells were divided into control group, Aβ25-35 group and BMSCs + Aβ25-35 group. The effects of BMSCs on PC12 cells treated by Aβ25-35 were detected using MTT, Hoechst 33258 and Annexin V-FITC/PI staining methods. The expression levels of TAG1, β-amyloid precursor protein (APP), AICD and p53 were determined by RT-PCR and Western blotting methods. The expression levels of Bax and Bcl-2 were determined by Western blotting method. The activity of Caspase 3 was detected by spectrophotometric method.
RESULTS
MTT results showed that cell activity decreased after the treatment of 20 μM Aβ25-35 for 48 h (P<0.01) while it increased in BMSCs + Aβ25-35 group (P<0.01). Hoechst 33258 and Annexin V-FITC/PI staining results showed that Aβ25-35 could induce the apoptosis of PC12 cells while the apoptosis of PC12 cells was inhibited in BMSCs + Aβ25-35 group. RT-PCR and Western blotting methods showed that 20 μM Aβ25-35 could increase the expression levels of TAG1, APP, AICD and p53 (P<0.01) while they decreased in BMSCs + Aβ25-35 group (P<0.01). 20 μM Aβ25-35 could increase the expression levels of Bax and decrease the expression levels of Bcl-2 (P<0.01), while the expression levels of Bax decreased and the expression levels of Bcl-2 increase in BMSCs + Aβ25-35 group (P<0.01). 20 μM Aβ25-35 could enhance Caspase 3 activity while it decreased in BMSCs + Aβ25-35 group (P<0.01). Conclusions BMSCs with Aβ25-35 could inhibit the apoptosis of PC12 cells, which maybe related with TAG1/APP/AICD signal pathway.
目的
本研究旨在探讨骨髓间充质干细胞（BMSCs）对瞬时轴突糖蛋白1（TAG1）介导的PC12细胞凋亡的保护作用。
方法
将PC12细胞分为对照组、Aβ25 - 35组和BMSCs + Aβ25 - 35组。采用MTT法、Hoechst 33258染色法及Annexin V - FITC/PI双染法检测BMSCs对Aβ25 - 35处理后的PC12细胞的影响。采用RT - PCR法和蛋白质免疫印迹法检测TAG1、β - 淀粉样前体蛋白（APP）、AICD和p53的表达水平。采用蛋白质免疫印迹法检测Bax和Bcl - 2的表达水平。采用分光光度法检测Caspase 3的活性。
结果
MTT结果显示，20 μM Aβ25 - 35处理48 h后细胞活性降低（P<0.01），而BMSCs + Aβ25 - 35组细胞活性升高（P<0.01）。Hoechst 33258染色及Annexin V - FITC/PI双染结果显示，Aβ25 - 35可诱导PC12细胞凋亡，而BMSCs + Aβ25 - 35组可抑制PC12细胞凋亡。RT - PCR法和蛋白质免疫印迹法显示，20 μM Aβ25 - 35可使TAG1、APP、AICD和p53的表达水平升高（P<0.01），而BMSCs + Aβ25 - 35组表达水平降低（P<0.01）。20 μM Aβ25 - 35可使Bax表达水平升高，Bcl - 2表达水平降低（P<0.01），而BMSCs + Aβ25 - 35组Bax表达水平降低，Bcl - 2表达水平升高（P<0.01）。20 μM Aβ25 - 35可增强Caspase 3活性，而BMSCs + Aβ25 - 35组活性降低（P<0.01）。结论：BMSCs联合Aβ25 - 35可抑制PC12细胞凋亡，其机制可能与TAG1/APP/AICD信号通路有关。
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