细胞共培养条件下大鼠肌源性干细胞对氧化应激损伤神经元的保护作用  

The protective effects of rats muscle-derived stem cells on neurons damage induced by oxidativestress in a co-culture system

作　　者：梅晰凡[1] 刘世琼[2] 刘畅[3] 董娜[4] 袁亚江[1] 郭占鹏[1] 李全双[1] 

机构地区：[1]辽宁医学院附属第一医院骨科,锦州121001 [2]河南科技大学附属第一医院骨科,洛阳471003 [3]辽宁医学院附属第一医院内分泌科,锦州121001 [4]河南科技大学附属第一医院病理科,洛阳471003

出　　处：《神经解剖学杂志》2011年第6期658-664,共7页Chinese Journal of Neuroanatomy

基　　金：辽宁省自然科学基金(20072204);辽宁省创新团队项目基金(2008T113)

摘　　要：目的:研究肌源性干细胞(MDSCs)与氧化应激损伤神经元共培养条件下MDSCs对后者的保护作用。方法:分离新生SD大鼠的MDSCs及皮层神经元,以叔丁基过氧化氢损伤神经元,构建氧化应激损伤的神经元模型。实验分为2组,损伤组:将第5代MDSCs和损伤的神经元置于共培养系统中培养4 h;对照组:损伤神经元于共培养系统中单独培养4 h。培养7 d后通过Hoster33258及流式细胞仪检测第2、4、6 d各组神经元的存活率,并用RT-PCR对相关基因进行分析,探究差异的内在原因。结果:Hoster33258及流式细胞仪检测结果显示:在共培养2、4、6 d三个时间点损伤组的神经元的存活率要远远高于对照组(P<0.05),凋亡率明显低于对照组,RT-PCR检测到损伤组神经元在三个时间点Bcl-2的表达水平均较对照组强(P<0.05),而Bax的表达水平均较对照组低(P<0.05)。结论:在共培养条件下,MDSCS对氧化应激损伤神经元具有保护作用,而这与其可促使损伤神经元Bcl-2的表达水平升高以及抑制Bax的表达有关。Objective： To study the protedive effects of muscle-derived stem cells（MDSCs） on neuronal damage induced by oxidative stress in a co-culture system.Methods： MDSCs and cortical neurons were isolated from newborn SD rats,oxidative stress neuronal damage model was induced by tert-butyl hydroperoxide in vitro.The co-cultured cells were divided into two groups randomly.Injury group： tert-butyl hydroperoxide treated group,the MDSCs of passage 5 were co-cultured with neurons which were treated with tert-butyl hydroperoxide for 4 hours.Contral group： the neurons which were treated with tert-butyl hydroperoxide for 4 hours were cultured along in the co-culture system.The neuronsal survival rate which were treated with tert-butyl hydroperoxide between co-cultured group and control group was detected by Hoster33258 and flow cytometry and RT-PCR at the 2nd,4th and 6th days and analyze the related genes by RT-PCR to explore the underlying causes of the difference.Results： The neuronal survival rate of injury group were significantly higher than the contral group by Hoster33258 and flow cytometry at the 2nd,4th and 6th days（P0.05）,RT-PCR results showed that Bcl-2 expressional level at three time in the neurons in injury groups was higher than control group（P0.05） and Bax expressional level was lower than control group（P0.05）.Conclusion： MDSCs could protect neurons against cells damage induced by oxidative stress possibly by up-regulating the expression of Bcl-2 mRNA,and the same time inhibit the expression Bax mRNA.

关 键 词：肌源性干细胞 细胞培养 BCL-2 BAX 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]