外泌体对脑缺血/再灌注损伤后星形胶质细胞炎症和MAPK-Cx43信号通路的影响  

Effects of Exosomes on Astrocyte Inflammation and MAPK-Cx43 Signaling after Cerebral IschemiaReperfusion Injury

作　　者：魏小兵[1] 温一奇 李素美 胡永珍 刘洋[1] 龙青山[1] 黄楚涛 刘惜弟 WEI Xiao-bing;WEN Yi-qi;LI Shu-mei;HU Yong-zhen;LIU Yang;LONG Qing-shan;HUANG Chu-tao;LIU Xi-di(Department of Neurosurgery,Huizhou Third People’s Hospital,Huizhou 516000,China)

机构地区：[1]惠州市第三人民医院神经外科,惠州516000

出　　处：《中国临床神经科学》2022年第1期16-22,55,共8页Chinese Journal of Clinical Neurosciences

摘　　要：目的探讨内皮细胞来源外泌体对脑缺血/再灌注(I/R)损伤后星形胶质细胞炎症和P38丝裂原活化蛋白激酶(P38 MAPK)-缝隙连接蛋白43(Cx43)信号通路的影响。方法分离人脐静脉内皮细胞来源的外泌体(HUVECExo)。将SD大鼠分为假手术组、大脑中动脉闭塞(MCAO)组、HUVEC-Exo治疗MCAO (HUVEC-Exo+MCAO)组,各组均n=10。其中,HUVEC-Exo+MCAO组按HUVEC-Exo干预浓度不同设为3个亚组(分别为10、20和30μg·mL;亚组),经立体定向微量注射器注入大鼠脑部。体外建立星形胶质细胞糖氧剥夺再灌注(OGD/R)模型,设为OGD/R组,对照组星形胶质细胞不进行处理。HUVEC-Exo+OGD/R组(HUVEC-Exo 10μg·mL^(-1)处理OGD/R星形胶质细胞24 h)。观察外泌体对I/R导致的大鼠脑梗死面积的影响。用ELISA和Western blot方法分析外泌体对OGD/R组的炎症因子和MAPK-Cx43蛋白表达的影响。结果与MCAO组比较,HUVEC-Exo+MCAO 10、20和30μg·mL^(-1)亚组的脑梗死面积和神经功能评分均明显降低(均P<0.05)。与OGD/R组比较,HUVEC-Exo+OGD/R组IL-6、IL-8、TNF-α表达下降(均P<0.05),MAPK-Cx43和磷酸化Cx43蛋白表达显著抑制(P<0.05)。结论 HUVEC-Exo可显著减少I/R大鼠的脑梗死面积,并且抑制OGD/R星形胶质细胞的炎症因子和MAPK-Cx43信号蛋白的表达。Aim To investigate the effects of endothelial cell-derived exosomes on cerebral ischemiareperfusion(I/R) rat, astrocyte inflammation and P38 mitogen-activated protein kinase(P38 MAPK)-connexin 43(Cx43) signal. Methods Exosomes were isolated from human umbilical vein endothelial cells(HUVEC-Exo). The rats were divided into a sham group, a I/R model by cerebral artery occlusion(MCAO group) and a MCAO treated with HUVEC-Exo group(HUVEC-Exo+MCAO group), with n=10 in each group. The exosomes of HUVEC-Exo+MCAO group were injected into the brain of MCAO group at 10, 20, and 30 μg·mL^(-1), respectively. The astrocyte oxygen-glucose deprivation reperfusion(OGD/R) model was established in vitro, and the astrocytes were not treated in the control group. In HUVECExo+OGD/R group, 10 μg· mL^(-1)HUVEC-Exo were treated with for 24 h in OGD/R cells, the effects of exosome on cerebral infarction size induced by I/R in rats were observed. The effects of exosomes on the expression of inflammatory factors and MAPK-Cx43 protein in OGD/R group were analyzed by ELISA and Western blot respectively. Results Compared with MCAO group, cerebral infarction area and neurological function score in HUVEC-Exo+MCAO groups of 10, 20, 30 μg·mL;were significantly decreased(all P<0.05). Compared with OGD/R group, the expressions of pro-inflammatory cytokines IL-6, IL-8 and TNF-α(P<0.05), map K-Cx43 and phosphorylated Cx43 proteins were significantly inhibited in HUVEC-Exo+OGD/R group(P<0.05). Conclusion The exosomes derived from endothelial cells can reduce the area of cerebral infarction and inhibit the expression of inflammatory factors in astrocytes, and the expression of MAPK-Cx43 signal protein.

关 键 词：脑 缺血/再灌注损伤 内皮细胞来源的外泌体 炎症 MAPK-Cx43信号通路 

分 类 号：R743[医药卫生—神经病学与精神病学] Q189[医药卫生—临床医学]