摘要
目的探讨白藜芦醇抑制Toll样受体4(TLR4)信号通路减轻脑缺血再灌注损伤后脑组织神经炎症的具体机制。方法将60只成年雄性SD大鼠按随机数字表法分为假手术组、脑缺血再灌注损伤组和白藜芦醇处理组,每组20只。后2组采用改良线栓法制备成短暂性大脑中动脉闭塞(MCAO)模型,闭塞2 h后进行再灌注,并分别于模型制备前15 min和再灌注前1 min给予20 mg/kg生理盐水或白藜芦醇尾静脉注射1次。于再灌注后72 h采用神经损伤严重性评分(NSS)评估各组大鼠的神经功能损伤程度,采用酶联免疫吸附试验检测大鼠损伤区脑组织内促炎因子白介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)和抗炎因子IL-4、IL-10、转化生长因子-β(TGF-β)的表达水平,采用逆转录-聚合酶链反应检测大鼠损伤区脑组织内小胶质细胞M1型特征性标志物IL-1βmRNA、CD32 mRNA和M2型特征性标志物CD206 mRNA、Arginase-1 mRNA的表达水平,采用Western blotting检测TLR4信号通路分子TLR4、髓样分化蛋白88(MyD88)、肿瘤坏死因子受体活化因子6(TRAF6)、IL-1R受体相关激酶1(IRAK1)和核因子-κB(NF-κB)蛋白的表达水平。结果与假手术组相比,脑缺血再灌注损伤组大鼠的NSS评分明显下降,损伤区脑组织内IL-1β、IL-6、TNF-α和IL-4、IL-10、TGF-β的表达水平均明显升高,IL-1βmRNA、CD32 mRNA和CD206 mRNA、Arginase-1 mRNA的表达水平均明显升高,TLR4、MyD88、TRAF6、IRAK1和NF-κB蛋白的表达水平均明显升高,差异均有统计学意义(P<0.05)。与脑缺血再灌注损伤组相比,白藜芦醇处理组大鼠的NSS评分明显下降,损伤区脑组织内IL-1β、IL-6、TNF-α的表达水平均明显降低,IL-4、IL-10、TGF-β的表达水平均明显升高,IL-1βmRNA、CD32 mRNA的表达水平均明显降低,CD206 mRNA、Arginase-1 mRNA的表达水平均明显升高,TLR4、MyD88、TRAF6、IRAK1和NF-κB蛋白的表达水平均明显降低,差异均有统计学意义(P<0.05)。结论白�
Objective To explore the mechanism of resveratrol in alleviating neuroinflammation after cerebral ischemia-reperfusion injury by inhibiting Toll like receptor 4(TLR4)signaling pathway.Methods Sixty adult male SD rats were randomly divided into sham-operated group,cerebral ischemia-reperfusion injury group,and resveratrol treatment group(n=20).Rat models of transient middle cerebral artery occlusion(MCAO)in the latter two groups were prepared by modified thread embolization method;reperfusion was performed after 2 h of occlusion,and 20 mg/kg normal saline or resveratrol via tail vein injection was given 15 min before model preparation and one min before reperfusion,respectively.At 72 h after MACO,neurological severity scale(NSS)was applied to evaluate the neurological functions of rats.Enzyme linked immunosorbent assay was performed to detect the expressions of pro-inflammatory cytokines interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-αand anti-inflammatory cytokines IL-4,IL-10,and transforming growth factor(TGF)-β.Reverse transcription-polymerase chain reaction was performed to detect the mRNA expressions of M1 signature markers IL-1βand CD32 and M2 signature markers CD206 and Arginase-1.Western blotting was used to detect the expressions of TLR4 signaling molecules TLR4,myeloid differentiation protein 88(MyD88),tumor necrosis factor receptor-associated factor 6(TRAF6),IL-1 receptor associated kinase 1(IRAK1)and nuclear factor(NF)-κB.Results As compared with those in the sham-operated group,rats in the cerebral ischemia-reperfusion injury group had significantly decreased NSS scores,significantly elevated levels of IL-1β,IL-6,TNF-α,IL-4,IL-10,and TGF-βin the damaged brain tissues,significantly elevated mRNA expression levels of IL-1β,CD32,CD206,and Arginase-1,and significantly elevated protein expression levels of TLR4,MyD88,TRAF6,IRAK1 and NF-κB(P<0.05).As compared with rats in the cerebral ischemia-reperfusion injury group,rats in the resveratrol treatment group had significantly decreased NSS scor
作者
马姗
闫忠军
李军朝
范玲玲
Ma Shan;Yan Zhongjun;Li Junchao;Fan Lingling(Department of Neurology,First Affiliated Hospital of Xi'an Medical College,Xi'an 710077,China;Department of Neurosurgery,Tangdu Hospital,Airforce Military Medical University,Xi'an 710038,China)
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2019年第11期1081-1086,共6页
Chinese Journal of Neuromedicine
基金
陕西省教育厅基金(18JK0661)。
