骨髓间充质干细胞对大鼠脑缺血再灌注损伤后神经元凋亡及炎症的影响  

Effects of bone marrow mesenchymal stem cells on neuronal apoptosis and inflammatory after cerebral ischemia-reperfusion injury in rats

作　　者：尚羽 李康睿 叶民[1] 沈华超 Shang Yu;Li Kangrui;Ye Min;Shen Huachao(Department of Neurology,BenQ Affiliated Hospital of Nanjing Medical University,Nanjing Jiangsu 210000,China;Department of Neurology,East War Zone General Hospital,Nanjing Jiangsu 210000,China)

机构地区：[1]南京医科大学附属明基医院神经内科,江苏南京210000 [2]东部战区总医院神经内科,江苏南京210000

出　　处：《遵义医科大学学报》2022年第3期289-295,共7页Journal of Zunyi Medical University

基　　金：南京市卫生科技发展专项基金项目(NO:YKK19144)。

摘　　要：目的探讨骨髓间充质干细胞在缺血性脑损伤中神经保护作用及其分子机制。方法将雄性SD大鼠随机分为3组:假手术组(Sham组)、模型组(MCAO组)、骨髓间充质干细胞组(BMSCs组)。除Sham组外,其他组构建大鼠大脑中动脉闭塞模型。神经功能缺损评分检测大鼠的神经功能情况;TTC染色法检测脑梗死体积;TUNEL染色检测神经元凋亡情况;酶联免疫吸附法(ELISA)检测血清中炎症因子TNF-α、IL-1β、IL-6和脑组织超氧化物歧化酶(SOD)和丙二醛(MDA)的表达;Western blotting检测Bax、Bcl-2、caspase-3、TLR4、NF-κB p65、NF-κB p50、IκB蛋白的表达。结果与Sham组比较,MCAO组大鼠mNSS评分、脑梗死体积均升高,血清中TNF-α、IL-1β和IL-6水平升高,缺血脑组织中SOD活性降低,MDA水平升高,脑组织凋亡细胞数量显著升高,Bax、caspase-3、TLR4、NF-κB p65、NF-κB p50蛋白表达均升高,Bcl-2、IκB蛋白表达降低,差异均有统计学意义(P<0.05);与MCAO组比较,BMSCs组大鼠mNSS评分、脑梗死体积均显著降低,血清中TNF-α、IL-1β、IL-6水平显著降低,缺血脑组织中SOD活性显著升高,MDA水平降低,脑组织凋亡细胞数量显著减少,Bax、caspase-3、TLR4、NF-κB p65、NF-κB p50蛋白表达均显著降低,Bcl-2、IκB蛋白表达显著升高,差异均有统计学意义(P<0.05)。结论BMSCs通过抗炎和抗凋亡作用对缺血性脑卒中发挥神经保护作用,其机制可能与抑制TLR4/NF-κB信号通路有关。Objective To investigate the neuroprotective effect and molecular mechanism of bone marrow mesenchymal stem cells in ischemic brain injury.Methods Male SD rats were randomly divided into 3 groups:sham operation(Sham),model(MCAO),bone marrow mesenchymal stem cell treatment(BMSCs).Except for the Sham group,other groups were subjected to middle cerebral artery occlusion.The modified neurological severity score(mNSS)was used to detect the neurological function of rats.TTC staining was used to detect cerebral infarction volume.TUNEL staining was used to detect apoptosis.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum inflammatory factors TNF-α,IL-1β,IL-6 and the expression of superoxide dismutase(SOD)and malondialdehyde(MDA)in brain tissue.Western blotting was used to detect the expression of Bax,Bcl-2,caspase-3,TLR4,NF-κB p65,NF-κB p50,and IκB protein.Results Compared with the Sham group,the mNSS score,the cerebral infarction volume,and serum TNF-α,IL-1βand IL-6 were increased in the MCAO group.The activity of SOD was decreased in the ischemic brain tissue,while the levels of MDA and apoptotic cells in brain tissue were increased in the MCAO group.The protein expressions of Bax,caspase-3,TLR4,NF-κB p65,and NF-κB p50 were increased,while the protein expressions of Bcl-2 and IκB were decreased in the MCAO group(P<0.05).Compared with the MCAO group,the mNSS score,the cerebral infarction volume,and serum levels of TNF-α,IL-1β,and IL-6 were significantly decreased in the BMSCs group.The activity of SOD in ischemic brain tissue was increased,while the level of MDA was decreased and apoptotic cells in brain tissue were reduced in the BMSCs group.The protein expressions of Bax,caspase-3,TLR4,NF-κB p65,and NF-κB p50 were decreased,while the protein expressions of Bcl-2 and IκB were increased in the BMSCs group(P<0.05).Conclusion BMSCs play a neuroprotective role in ischemic stroke through anti-inflammatory and anti-apoptotic effects,which may be related to inhibition of TLR4/NF-κB signaling pa

关 键 词：大鼠 骨髓间充质干细胞 缺血性脑损伤 TLR4/NF-κB 凋亡 炎症 

分 类 号：R743.3[医药卫生—神经病学与精神病学]