黄芪甲苷抑制Fas/FasL信号通路减轻创伤性脑损伤大鼠神经功能缺损和神经元凋亡  

Astragaloside inhibits Fas/FasL signaling pathway to reduce neural dysfunction and neuronal apoptosis in traumatic brain injury of rats

作　　者：陈惠刚 池小锋 封娣 米娅莉 CHEN Huigang;CHI Xiaofeng;FENG Di;MI Yali(Clinical Nursing Teaching and Research Office of the School of Nursing at Zhangjiakou University,Zhangjiakou 075061,China)

机构地区：[1]张家口学院护理学院临床护理教研室,075061

出　　处：《天津医药》2024年第5期469-474,共6页Tianjin Medical Journal

基　　金：河北省医学科学研究课题计划项目(20220572)。

摘　　要：目的探究黄芪甲苷通过Fas/FasL信号通路对创伤性脑损伤(TBI)大鼠神经功能缺损和神经元凋亡的影响。方法将大鼠随机分为5组:假手术组、模型组、黄芪甲苷组(20 mg/kg)、Fas沉默组[4µg Fas小干扰RNA(siRNA)慢病毒载体]、黄芪甲苷+Fas沉默组(20 mg/kg黄芪甲苷+4µg Fas siRNA慢病毒载体),每组10只。除假手术组外,其余各组大鼠制作TBI模型。各组按照对应剂量进行给药干预,每日1次,持续7 d。水迷宫实验检测大鼠神经功能缺损;荧光定量PCR法检测脑组织Fas、FasL mRNA表达;苏木精-伊红、β-微管蛋白Ⅲ(Tuj1)免疫荧光、TUNEL染色分别观察脑组织病理变化、神经元活性及神经元凋亡;Western blot法检测脑组织中Fas/FasL通路、胱天蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达。结果与假手术组比较,模型组大鼠脑组织细胞间隙增加、变性明显,第1~5天大鼠逃避潜伏期,神经元凋亡率,Caspase-3、Bax蛋白,Fas、FasL mRNA和蛋白水平升高,穿越平台次数、Tuj1阳性细胞数目、Bcl-2蛋白水平降低(P<0.05)。与模型组比较,黄芪甲苷组、Fas沉默组大鼠脑组织细胞间隙缩小,变性减轻,第1~5天大鼠逃避潜伏期,神经元凋亡率,Caspase-3、Bax蛋白,Fas、FasL mRNA和蛋白水平降低,穿越平台次数、Tuj1阳性细胞数目、Bcl-2蛋白水平升高(P<0.05)。黄芪甲苷组和Fas沉默组大鼠脑组织病理变化和上述指标差异无统计学意义(P>0.05)。与黄芪甲苷组和Fas沉默组比较,黄芪甲苷+Fas沉默组大鼠脑组织病理损伤进一步降低,神经元凋亡率,Caspase-3、Bax蛋白,Fas、FasL mRNA和蛋白水平降低,Tuj1阳性细胞数目、Bcl-2蛋白水平升高(P<0.05)。结论黄芪甲苷通过抑制Fas/FasL信号通路介导的凋亡途径减轻TBI大鼠神经功能缺损和神经元凋亡。Objective To explore the effects of astragaloside on neural dysfunction and neuronal apoptosis of traumatic brain injury(TBI)in rats through Fas/FasL signaling pathway.Methods Fifty rats were randomly divided into 5 groups:the sham operation group,the model group,the astragaloside group(20 mg/kg),the Fas silencing group[4µg Fas small interfering RNA(siRNA)lentiviral vector]and the astragaloside+Fas silencing group(20 mg/kg astragaloside+4µg Fas siRNA lentiviral vector).Each group consisted of 10 rats.Except the sham operation group,the other groups of rats were established TBI rat model.Each group was received medication intervention according to the corresponding dosage,once a day,for 7 days.Water maze test was used to detect the nerve function defect in rats.Fluorescence quantitative PCR was used to detect expression levels of Fas and FasL messenger RNA(mRNA)in brain tissue.Hematoxylin-eosin,β-tubulin III(Tuj1)immunofluorescence and TUNEL staining were used to observe pathological changes,neuronal activity and apoptosis of brain tissue,respectively.Western blot assay was used to detect expression levels of Fas/FasL pathway,Caspase-3,Bcl-2 associated X protein(Bax)and B lymphoblastoma-2(Bcl-2)protein in brain tissue.Results Compared with the sham operation group,the cell gap was increased and degeneration was obvious in brain tissue in the model group.The escape latency of rats on days 1-5,neuronal apoptosis rate,Caspase-3,Bax protein,Fas,FasL mRNA and protein levels were increased(P<0.05).The number of times crossing platforms,the number of Tuj1 positive cells and Bcl-2 protein were decreased(P<0.05).Compared with the model group,in the astragaloside group and the Fas silencing group,the cell gap in brain tissue was narrowed and degeneration was reduced,the escape latency of rats on days 1-5,neuronal apoptosis rate,Caspase-3,Bax protein,Fas,FasL mRNA and protein levels were decreased,and the number of crossing platforms,the number of Tuj1 positive cells and Bcl-2 protein level were increased(P<0.05).There we

关 键 词：脑损伤 创伤性 黄芪甲苷 FAS受体 神经元 

分 类 号：R651.15[医药卫生—外科学] R286.1[医药卫生—临床医学]