摘要
为探究骨髓间充质干细胞(bone marrow meaenchymal stem cells,BMSCs)源性外泌体(exosomes,Exos)对急性期脑缺血大鼠小胶质细胞/巨噬细胞M1/M2极化的影响,采用超高速离心法分离提取外泌体并鉴定;采用线栓法制备大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)模型;采用Longa评分和角实验评价大鼠神经功能,2,3,5-氯化三苯基四氮唑(2,3,5-triphenyltetrazole chloride,TTC)染色检测大鼠脑梗死体积;采用CD16/32/Iba1、CD206/Iba1免疫荧光双标法检测小胶质细胞/巨噬细胞M1/M2表型;采用RT-qPCR检测大鼠脑缺血周边区CD86、诱导型一氧化氮合酶(inducible nitricoxide synthase,iNOS)、肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、精氨酸酶1(arginase-1,Arg-1)、白细胞介素-10(interleukin-10,IL-10)和转化生长因子β(transforming growth factor beta,TGF-β)的mRNA表达。实验结果表明,BMSC-Exos减少缺血周边区CD16/32+/Iba1+阳性细胞数量(P<0.01),增加CD206+/Iba1+阳性细胞数量(P<0.01),减少iNOS、CD86和TNF-α的mRNA表达,增加Arg-1、TGF-β和IL-10的mRNA表达(P<0.05或P<0.01)。本研究提示BMSC-Exos调控急性期脑缺血大鼠小胶质细胞/巨噬细胞M1/M2极化,减轻神经炎症,改善脑缺血损伤。
The aim of the present study was to investigate the effects of exosomes derived from bone marrow mesenchymal stem cells(BMSCs)on the polarization of M1/M2 microglia/macrophages in rats with acute cerebral ischemia.Ultrahigh-speed centrifugation was employed to isolate and identify exosomes;a middle cerebral artery occlusion(MCAO)model was prepared in rats using the intraluminal filament technique;Longa scoring and corner tests were used to evaluate the neurological function of rats;2,3,5-triphenyltetrazole chloride(TTC)staining was used to assess the infarct volume in rat brains;immunofluorescence double-labeling of CD16/32/Iba1 and CD206/Iba1 was performed to detect M1/M2 phenotypes of microglia/macrophages;RT-qPCR was employed to measure the mRNA expression of CD86,inducible nitric oxide synthase(iNOS),tumor necrosis factor-alpha(TNF-α),arginase-1(Arg-1),interleukin-10(IL-10),and transforming growth factor beta(TGF-β)in the ischemic penumbra of rat brains.The experimental results showed that BMSC-Exos reduced the number of CD16/32+/Iba1+positive cells in the ischemic penumbra(P<0.01)while increasing the number of CD206+/Iba1+positive cells(P<0.01),and decreased the mRNA expression of iNOS,CD86,and TNF-α,while increasing the mRNA expression of Arg-1,TGF-β,and IL-10(P<0.05 or P<0.01).This research suggests that BMSC-Exos can regulate M1/M2 polarization of microglia/macrophages in rats with acute cerebral ischemia,alleviate neuroinflammation,and improve ischemic brain injury.
作者
孙逸梅
毛诗慧
李琳
江伟锋
储利胜
SUN Yimei;MAO Shihui;LI Lin;JIANG Weifeng;CHU Lisheng(School of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China)
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2023年第5期599-606,共8页
Journal of China Pharmaceutical University
基金
国家自然科学基金资助项目(No.81274113,No.81873028)
浙江省公益技术应用研究计划资助项目(No.2016C33185)。
