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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年06期

页码:

522-527

栏目:

基础医学

出版日期:

2024-11-20

文章信息/Info

Title:

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

文章编号:

1006-8147(2024)06-0522-06

作者:

吴慧娴; 练学淦

（江苏省常州市第一人民医院神经内科，常州213003）

Author(s):

WU Huixian; LIAN Xuegan

（Department of Neurology，Changzhou First People′s Hospital，Changzhou 213003，China）

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

Keywords:

bone marrow mesenchymal stem cells;  interleukin-12;  hypoxia;  ischemia reperfusion;  inflammatory

分类号:

R743.31

DOI:

10.20135/j.issn.1006-8147.2024.06.0522

文献标志码:

A

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。

Abstract:

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. According to the RT-PCR results，the mRNA expression of JAK （F=14.83，P<0.01） and STAT4 （F=37.95，P<0.05） was reduced in comparison to the MCAO group. Conclusion：BMSCs have neuroprotective effects on ischemic stroke mouse models，and regulating IL-12 expression and its mediated JAK/STAT4 signaling pathway may be the molecular mechanism for improving the inflammatory response in ischemic injury areas.

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相似文献/References:

备注/Memo

备注/Memo:

作者简介 吴慧娴（1998-），女，硕士在读，研究方向：脑血管病；
通信作者：练学淦，E-mail：lxgneuro@126.com。

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更新日期/Last Update: 2024-11-25