摘要
[目的]研究依达拉奉右莰醇(ED)对液压冲击脑损伤大鼠小胶质细胞极化的影响,并基于Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路探讨其机制。[方法]从205只健康雄性SD大鼠中随机取32只设为假手术组,其余173只大鼠采用液压冲击法制备脑损伤大鼠模型,取160只成模大鼠随机分为模型组、ED(7 mg/kg)组、TAK242(瑞沙托维,TLR4抑制剂,2 mg/kg)组、ED(7 mg/kg)+TAK242(2 mg/kg)组和ED(7 mg/kg)+脂多糖(LPS,TLR4激动剂,0.4 mg/kg)组,每组32只。各组分别1次/天连续腹腔注射给药14天后,通过改良神经功能缺损评分(mNSS)、失重法、伊文思蓝(EB)渗透法考察大鼠神经功能、脑含水量和血-脑屏障(BBB)通透性,通过HE染色、Nissl染色考察脑组织病理学改变,ELISA检测脑组织γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α)、白细胞介素(IL)1β、IL-4、IL-10水平,免疫荧光双染法检测小胶质细胞M1极化表型(CD86/Iba-1)和M2极化表型(CD206/I ba-1),RT-PCR、Western blot检测TLR4、NF-κB p65、NOD样受体蛋白3(NLRP3)、水通道蛋白4(AQP4)mRNA和蛋白表达。[结果]与模型组相比,ED组、TAK242组和ED+TAK242组大鼠mNSS评分、脑含水量、BBB通透性明显降低(P<0.05),脑组织结构紊乱、神经元排列稀疏无序、空泡样变、炎症细胞浸润、尼氏小体数量减少等病理学改变明显改善,脑组织IFN-γ、TNF-α、IL-1β水平明显降低,IL-4、IL-10水平明显升高(P<0.05),小胶质细胞M1极化表型阳性细胞率明显降低,M2极化表型阳性细胞率明显升高(P<0.05),TLR4、NF-κB p65、NLRP3、AQP4的mRNA和蛋白表达量均明显降低(P<0.05)。TAK242可明显增强ED对液压冲击脑损伤大鼠神经功能、脑含水量、BBB通透性、炎症反应、小胶质细胞极化、TLR4/NF-κB信号通路相关mRNA和蛋白表达的调控作用(P<0.05),LPS则明显逆转ED对液压冲击脑损伤大鼠的上述调控作用(P<0.05)。[结论]ED可能通过抑制TLR4/NF-κB信号通路而促进小胶质细�
Aim To investigate the effect of edaravone dexborneol(ED)on microglial polarization in rats with brain injury caused by hydraulic shock,and explore its mechanism based on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.Methods 32 rats of 205 healthy male SD rats were randomly selected as sham group,the remaining 173 rats were prepared with brain injury model by hydraulic shock method,and 160 model rats were randomly divided into model group,ED(7 mg/kg)group,TAK242(ressatovir,TLR4 inhibitor,2 mg/kg)group,ED(7 mg/kg)+TAK242(2 mg/kg)group and ED(7 mg/kg)+lipolyaccharide(LPS,TLR4 agonist,0.4 mg/kg)group,with 32 rats in each group.After 14 days of continuous intraperitoneal injection once a day,the nerve function,brain water content and blood-brain barrier(BBB)permeability were measured by modified neurological severity score(mNSS),weightlessness method or Evans blue(EB)penetration method,the brain histopathological changes was observed by HE and Nissl staining,the levels of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleukin(IL)1β,IL-4 and IL-10 in brain tissue were detected by ELISA,the M1 polarization phenotype(CD86/Iba-1)and M2 polarization phenotype(CD206/I ba-1)of microglia cells were detected by immunofluorescence double staining,the mRNA and protein expressions of TLR4,NF-κB p65,NOD-like receptor protein 3(NLRP3),aquaporin 4(AQP4)were detected by RT-PCR or Western blot.Results Compared with the model group,the mNSS score,brain water content,BBB permeability of the rats in ED group,TAK242 group,ED+TAK242 group were significantly were decreased(P<0.05),the pathological changes such as brain structure disorder,sparse and disordered neuronal arrangement,vacuole-like transformation,inflammatory cell infiltration,decrease in the number of Nishi bodies were significantly improved,the levels of IFN-γ,TNF-α,IL-1βin brain tissue were significantly decreased,while the levels of IL-4,IL-10 were significantly increased(P<0.05),the m RNA and protein expressions of TLR4,NF-κB p65,N
作者
徐伟
董雨
申向竹
杨德真
唐会昌
武一平
XU Wei;DONG Yu;SHEN Xiangzhu;YANG Dezhen;TANG Huichang;WU Yiping(The Fourth Department of Neurosurgery,Handan Central Hospital,Handan,Hebei 056001,China)
出处
《中国动脉硬化杂志》
CAS
2024年第2期109-117,共9页
Chinese Journal of Arteriosclerosis
基金
河北省医学科学研究课题(20220516)。
