hUC-MSCs对缺氧缺血脑损伤新生大鼠脑组织的保护作用研究  

Protective effect of hUC-MSCs on brain tissue of neonatal rats with hypoxic-ischemic brain injury

作　　者：陈茂琼 向敏[1,2,3] 宋海良 王岚 匡梦岚[2,3,6] 张丽娟 熊英 何志旭 许键炜 高鸿[9] 秦臻 Chen Maoqiong;Xiang Min;Song Hailiang(Dept of Neonatal Pediatrics, Affiliated Hospital of Guizhou Medical University, Guiyang 550004;National Local Joint Engineering Laboratory of Cell Engineering Biopharmaceutical Technology, Tissue Engineering and Stem Cell Experimental Center, Guizhou Medical University, Guizhou Key Laboratory of Regenerative Medicine, Guiyang 550000;Key Experiment on The Transformation of Adult Stem Cells of The Chinese Academy of Medical Sciences, Guiyang 550004;Dept of General Surgery, Dalang Hospital, Dongguan City, Dongguan 523770)

机构地区：[1]贵州医科大学附属医院新生儿科,贵阳550004 [2]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,组织工程与干细胞实验中心,贵州省再生医学重点实验室,贵阳550000 [3]中国医学科学院成体干细胞转化研究重点实验,贵阳550004 [4]东莞大朗医院普外科,东莞523770 [5]贵州医科大学医学科学研究所,贵阳550000 [6]贵州医科大学基础医学院药理学教研室,贵阳550000 [7]贵州医科大学护理学院,贵阳550000 [8]贵州医科大学基础医学院机能学实验室,贵阳550000 [9]贵州医科大学第三附属医院麻醉科,都匀558000

出　　处：《安徽医科大学学报》2021年第8期1226-1231,共6页Acta Universitatis Medicinalis Anhui

基　　金：中国医学科学院中央级公益性科研院所基本科研业务费专项资金(编号:2018PT31048);贵州省科技厅计划项目(编号:黔科合支撑[2017]2873);贵州省科技厅计划项目(编号:黔科合LH字[2016]7361)。

摘　　要：目的通过移植人脐带来源的间充质干细胞(hUC-MSCs)治疗缺氧缺血诱导的新生大鼠缺氧缺血性脑病(HIE),并探讨hUC-MSCs对神经细胞的保护作用及机制。方法采集足月健康新生儿脐带约10 cm,运用组织块贴壁法培养hUC-MSCs,通过感染重组携带绿色荧光蛋白的腺病毒标记细胞;取18只新生健康7d龄SD大鼠,随机均分为对照(假手术)、模型、治疗组,其中模型组及治疗组建立新生大鼠缺氧缺血脑损伤模型,建模成功24 h后将标记的hUC-MSCs注射入治疗组左侧(损伤侧)侧脑室,各组大鼠hUC-MSCs移植治疗48 h后,检测hUC-MSCs在HIE大鼠大脑内的定植并采用TTC染色观察梗死脑组织,RT-qPCR测定海马组织中Beclin-2、Caspase-3 mRNA水平的变化,Western blot检测海马区细胞Beclin-2、Caspase-3蛋白的表达。结果对照组大鼠大脑没有形成梗死体积,模型组大鼠大脑梗死体积明显,梗死体积比为(41.67±4.17)%;治疗组大鼠经hUC-MSCs移植治疗48 h后,大脑梗死体积减小[(16.65±3.43)%],与模型组比较,差异有统计学意义;HIE建模48 h后,内源性Beclin-2、Caspase-3的mRNA以及蛋白的表达上调。hUC-MSCs移植治疗48 h后,Beclin-2、Caspase-3的mRNA以及蛋白的表达下降。结论hUC-MSCs能减小缺氧缺血损伤大鼠大脑梗死体积,对损伤大鼠有保护作用;hUC-MSCs能够减少大鼠脑细胞凋亡,下调缺氧缺血脑损伤大鼠海马区细胞凋亡相关Beclin-2、Caspase-3的mRNA以及蛋白水平。Objective To explore the protective effect and mechanism of human umbilical cord derived mesenchymal stem cells(hUC-MSCs)on neurons by transplanting hUC-MSCs and treating hypoxic ischemic encephalopathy(HIE)in neonatal rats induced by hypoxia and ischemia.Methods About 10 cm umbilical cord of normal full-term healthy newborns born was collected under sterile conditions,and hUC-MSCs were cultured by tissue block adherent method,and then infected with recombinant adenovirus carrying green fluorescent protein.18 healthy7-day-old SD rats were randomly divided into normal control(sham operation)group,model group and hUC-MSCs treatment group,6 in each group.After 24 h of modeling,hUC-MSCs were injected into the left ventricle of the treatment group.After 48 h of hUC-MSCs transplantation,hUC-MSCs were detected in the brain of HIE rats and TTC staining was used to observe the infarcted brain tissue.The mRNA levels of Beclin-2 and Caspase-3 in hippocampus were detected by RT-qPCR.Western blot was used to detect the expression of Beclin-2 and Caspase-3.Results 48 h after hUC-MSCs transplantation in HIE group,there was no infarct volume in normal control group,and the infarct volume ratio was(41.67±4.17)%;in hUC-MSCs treatment group,the infarct volume ratio was(16.65±3.43)%,which was significantly lower than that in hypoxic-ischemic brain injury group.48 h after HIE modeling,the expression of endogenous beclin-2 mRNA and caspase-3 mRNA,beclin-2 protein and caspase-3 protein were significantly up-regulated.48 h after hUC-MSCs transplantation,the expression of beclin-2 mRNA,caspase-3 mRNA,beclin-2 protein and caspase-3 protein decreased.Conclusion hUC-MSCs can effectively reduce the infarct volume of rats with hypoxic-ischemic injury,which has a protective effect on the injured rats;at the same time,hUC-MSCs can significantly reduce the apoptosis of rat brain cells,and significantly down regulate the levels of beclin-2 mRNA,caspase-3 mRNA,beclin-2 protein and caspase-3 protein in hippocampus of rats with hypoxic-ischemic br

关 键 词：缺氧缺血性脑病 人脐带来源的间充质干细胞 凋亡 Beclin-2 Caspase-3 

分 类 号：R22[医药卫生—中医基础理论]