骨髓基质干细胞移植改善慢性酒精中毒大鼠脑损害的相关机制研究  

Effect and Mechanism of Intravenous Bone Marrow Stromal Cells Transplantation Against Chronic Ethanol-induced Brain Damage in Rats

作　　者：刘蕾[1] 胡建[1] 董凤[1] 徐晔[1] 夏炎[1] 吴铮[1] LIU Lei;HU Jian;DONG Feng;XU Ye;XIA Yan;WU Zheng(Department of Psychiatry,The First Affiliated Hospital,Harbin Medical University,Harbin,Heilongjiang,150001,China)

机构地区：[1]哈尔滨医科大学附属第一医院精神卫生中心

出　　处：《现代生物医学进展》2019年第11期2041-2045,共5页Progress in Modern Biomedicine

基　　金：黑龙江省教育厅科学技术研究项目(面上项目)(12511291)

摘　　要：目的:探讨骨髓基质干细胞(bone marrow stormal cells, BMSCs)静脉移植对慢性酒精中毒大鼠脑保护作用的相关机制。方法:体外分离、培养、扩增SD大鼠BMSCs。成年雄性SD大鼠随机分为慢性酒精中毒组、BMSCs回输组、磷酸缓冲盐溶液(phosphate buffer saline,PBS)回输组和对照组,每组7只。前三组用酒精灌胃8周建立慢性酒精中毒动物模型,对照组不造模(给予蔗糖灌胃),BMSCs回输组和PBS回输组于造模7周时一次性经尾静脉回输BMSCs或PBS。免疫印迹法检测海马Bcl-2、Bax、NGF、BDNF以及信号转导分子p-Akt的表达;反转录PCR检测海马神经生长因子(nerve growth factor, NGF)和脑源性神经营养因子(brain derived neurotrophic factor, BDNF)。结果:BMSCs回输组海马抗凋亡蛋白Bcl-2表达高于其余三组(P<0.05);促凋亡蛋白Bax表达低于慢性酒精中毒组(P<0.01),与对照组无统计学差异(P=0.989)。BMSCs回输组鼠海马内NGF和BDNF m RNA和蛋白表达、p-Akt蛋白表达均高于其余三组(P<0.05)。结论:静脉移植BMSCs能够明显改善慢性酒精中毒大鼠海马的细胞凋亡;其可能与自或旁分泌BDNF和NGF营养因子有关,且可能部分是通过激活PI3K/Akt通路实现。Objective: To investigate the effect and mechanism of intravenous bone marrow stromal cells(BMSCs) transplantation against chronic ethanol-induced brain damage in rats. Methods: BMSCs from rats were isolated, cultured and identified in vitro. Male Sprague-Dawley rats(n=28) were randomly divided into four groups: chronic ethanol group, BMSCs group, PBS group and control group, BMSCs or PBS were intravenously injected to the rats of BMSCs or PBS group at the 7 th week after ethanol gavage, respectively.The expression of Bcl-2, Bax, and phospho-Akt in hippocampus were evaluated by immunoblot. Nerve growth factor(NGF) and brain derived neurotrophic factor(BDNF) protein and m RNA expression in hippocampus were analysed by immunoblot and reverse transcriptase-polymerase chain reaction(RT-PCR), respectively. Results: We observed that transplated BMSCs significantly reduced the Bax expression(P<0.01) and increased Bcl-2 expression(P<0.05) in hippocampus of ethanol-fed rats. We also found BMSCs transplantation was able to induce upregulaton of NGF of BDNF in hippocampus of ethanol-fed rats at m RNA and protein levels(P<0.05). In addition,we discovered that the phospho-Akt expression was increased in hippocampus of BMSCs transplated rats than that in other three groups(P<0.01). Conclusion: Intravenous BMSCs transplantation is able to shield rat cell apoptotic impairment from chronic alcoholism by autocrine/paracrine of neurotrophic factors, which is involved in PI3 K/Akt activation, at least partially.

关 键 词：骨髓基质干细胞 慢性酒精中毒 凋亡 神经营养因子 PI3K/AKT通路 

分 类 号：R-33[医药卫生] Q593.2[生物学—生物化学]