2,5-己二酮致大鼠骨髓间充质干细胞凋亡的研究

2,5-Hexanedione induces apoptosis inthemesenchymal stem cells from rat bone marrow

关键词:  2,5-己二酮  神经毒性  大鼠骨髓间充质干细胞  凋亡

Key words:  [Key words] 2,5-hexanedione  neurotoxicity  bone marrow mesenchymal stem cells (BMSCs)  apoptosis

摘要:

[摘要] 　目的　研究 2,5-己二酮（2,5-HD）对大鼠骨髓间充质干细胞（BMSCs）的凋亡作用及其机制。〖HTH〗方法〖HTK〗　从大鼠骨髓分离、提取和培养大鼠BMSCs，经形态学观察，流式检测特异细胞标记物表达，成骨、成脂细胞诱导分化，确认其为BMSCs。将BMSCs暴露于10 mmol/L,20 mmol/L,40 mmol/L 2,5-HD，培养24 h，并设立空白对照组。用MTT法观察细胞存活率；用HE染色法观察染毒后细胞形态变化；用Hoechst染色法检测细胞凋亡；用Western Blot检测Bax、Bcl-2蛋白表达水平。〖HTH〗结果　〖HTK〗2,5- HD染毒组BMSCs存活率显著低于对照组（P<0.05）。HE染色观察显示，染毒BMSCs胞体变小，胞核固缩，突起变短。Hoechst染色观察结果表明，染毒BMSCs可见核的固缩及新月形等凋亡特征的核形态变化。Western Blot结果表明，与对照组比较，染毒组BMSCs的Bax蛋白水平显著升高（P<0.05），Bcl-2蛋白水平显著下降（P<0.05）。〖HTH〗结论　〖HTK〗2,5-HD能诱导大鼠BMSCs凋亡，且其凋亡诱导机制可能与Bax/Bcl-2蛋白的表达紊乱有关。

Abstract:

[Abstract] Objective To investigate apoptotic effect of 2,5-Hexanedione (2,5-HD) on bone marrow mesenchymal stem cells (BMSCs) of rats and its mechanism. 　Methods Thecellswere isolated from Bone marrows of rats and cultured. The morphology ofcells were observed, their surface markers were determined by flow cytomerty. Moreover, the cells could be induced tofat cells andosteoblasts. These cells were confirmed to be BMSCs. In this study, the fifth generation of rat BMSCs was exposed to 0, 10, 20 and 40 mmol/L 2,5-HD for 24 h and the cells viability was measured by MTT assay. Themorphological changes of BMSCs were observed using HE staining and The apoptotic morphologic changes of BMSCs were observed by Hoechst 33342 staining. The protein expression of Bcl-2 and Baxwasdetermined by Western blot. 　Results Our results showed thatexposure to 2,5-HD decreased survival of BMSCs and induced apoptosis. Moreover, 2,5-HD significantly increasedBaxexpression(P<0.05)and significantly decreased Bcl-2 expression(P<0.05). 　Conclusion These results suggest that 2,5-HD-induced apoptosis of BMSCs may be associated withdisturbanceinBax/Bcl-2 protein expression.