低氧预处理骨髓间充质干细胞对小鼠脑缺血再灌注模型中IL-12的影响  

The effect of hypoxic pretreatment of bone marrow mesenchymal stem cells on IL-12 after cerebral ischemia-reperfusion injury in mice

作　　者：吴慧娴 练学淦[1] WU Huixian;LIAN Xuegan(Department of Neurology,Changzhou First People′s Hospital,Changzhou 213003,China)

机构地区：[1]江苏省常州市第一人民医院神经内科,常州213003

出　　处：《天津医科大学学报》2024年第6期522-527,共6页Journal of Tianjin Medical University

基　　金：江苏省级横向课题(JKHY2021005)。

摘　　要：目的:探讨骨髓间充质干细胞(BMSCs)对缺血性脑损伤后白细胞介素(IL)-12介导炎症的影响及其分子机制。方法:将雄性C57BL6小鼠随机分为3组:假手术组(Sham组)、模型组(MCAO组)、BMSCs组,每组20只。除Sham组外,其他组构建小鼠大脑中动脉闭塞模型。神经功能缺损评分评价小鼠的神经功能;3,5-氯化三苯基四氮唑(TTC)染色检测脑梗死体积,免疫荧光检测小胶质细胞活化情况,Western印迹检测IL-1β、肿瘤坏死因子(TNF)-α、IL-6、干扰素(IFN)-γ、IL-4、IL-12蛋白的表达。RT-PCR检测脑组织损伤区酪氨酸蛋白激酶(JAK)/信号转导与转录激活因子4(STAT4)mRNA的表达。结果:与Sham组相比,MCAO组小鼠Clark评分(t=18.27,P<0.001)、脑梗死面积均增加(F=41.18,P<0.001),脑组织TNF-α(F=13.81,P<0.01)、IL-1β(F=8.753,P<0.01)和IL-6(F=10.96,P<0.01)、IFN-γ(F=18.08,P<0.01)水平升高,IL-4(F=10.76,P<0.05)表达水平降低;与MCAO组相比,BMSCs组小鼠Clark评分(t=3.416,P<0.05)、脑梗死面积均显著降低(F=41.18,P<0.05),脑组织中TNF-α(F=13.81,P<0.05)、IL-1β(F=8.753,P<0.05)和IL-6(F=10.96,P<0.05)、IFN-γ(F=18.08,P<0.05)表达水平显著降低,IL-4(F=10.76,P<0.01)表达水平升高。Western印迹显示,BMSCs组较MCAO组脑组织中IL-12(F=9.927,P<0.05)含量明显下降。RT-PCR结果显示,JAK(F=14.83,P<0.01)、STAT4的mRNA表达(F=37.95,P<0.05)较MCAO组降低。结论:BMSCs对缺血性脑卒中小鼠模型有神经保护作用,调控IL-12的表达与其介导的JAK/STAT4信号通路可能是其改善缺血损伤区炎性反应的分子机制。Objective:To investigate the effect of bone marrow mesenchymal stem cells(BMSCs)on the interleukin(IL)-12-mediated inflammatory response and their molecular mechanisms following ischemic brain injury.Methods:The male C57BL6 mice were randomly divided into three groups:the sham surgery group(Sham group),the model group(MCAO group),and the BMSCs group,with 20 mice in each group.Except for the Sham group,all other groups established a middle cerebral artery occlusion model in mice.The neurological defect score was used to assess the neurological function of mice.The 3,5-triphenyltetrazolium chloride(TTC)staining method was used to measure the volume of cerebral infarction,and the microglial cell activation was detected by immunofluorescent detection.The expression of IL-1β,tumor necrosis factor-alpha(TNF-α),IL-6,interferon-γ(IFN-γ),IL-4,and IL-12 proteins were detected by Western blotting.RT-PCR was used to detect the expression of tyrosine protein kinase(JAK)/signal transducer and activator of transcription 4(STAT4)mRNA in brain tissue injury area.Results:Compared to the Sham group,the Clark score(t=18.27,P<0.001)and the cerebral infarction area in mice of the MCAO group were significantly increased(F=41.18,P<0.001).The levels of TNF-α(F=13.81,P<0.01),IL-1β(F=8.753,P<0.01),IL-6(F=10.96,P<0.01),and IFN-γ(F=18.08,P<0.01)in brain tissue were elevated,while the expression level of IL-4(F=10.76,P<0.05)was significantly decreased.In contrast,when compared to the MCAO group,the Clark score(t=3.416,P<0.05)and the cerebral infarction area in mice of the BMSCs group were significantly reduced(F=41.18,P<0.05).The expression levels of TNF-α(F=13.81,P<0.05),IL-1β(F=8.753,P<0.05),IL-6(F=10.96,P<0.05),and IFN-γ(F=18.08,P<0.05)in brain tissue were also significantly reduced,while the expression level of IL-4(F=10.76,P<0.01)was significantly increased.Western blotting analysis revealed that the content of IL-12(F=9.927,P<0.05)in brain tissue was significantly decreased in the BMSCs group compared to the MCAO group.Accordi

关 键 词：骨髓间充质干细胞 白细胞介素-12 低氧 缺血-再灌注 炎症 

分 类 号：R743.31[医药卫生—神经病学与精神病学]