3种神经细胞保护措施对缺氧缺血性脑损伤新生大鼠脑组织Fas/FasL表达的影响  

Effect of 3 neuroprotective measures on expressions of Fas/FasL in brain tissue of neonatal rats with hypoxicischemic brain damage

作　　者：张晶[1] 董燕[1] 胡宝玲 黄蕊[1] 赵倩[1] 张旭辉[3] Zhang Jing;Dong Yan;Hu Baoling;Huang Rui;Zhao Qian;Zhang Xuhui(Department of Pediatrics,the First Hospital of Hebei Medical University,Shijiazhuang 050031,Hebei,China;Department of Health Management,the First Hospital of Hebei Medical University,Shijiazhuang 050031,Hebei,China;Department of Pediatrics,Shijiazhuang First Hospital,Shijiazhuang 050011,Hebei. China)

机构地区：[1]河北医科大学第一医院儿科,河北石家庄050031 [2]河北医科大学第一医院健康管理部,河北石家庄050031 [3]石家庄市第一医院儿科,河北石家庄050011

出　　处：《中国中西医结合急救杂志》2019年第1期11-15,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：河北省重点研发计划项目(182777109D);河北医科大学第一医院“星火”青年科研项目(XH201712).

摘　　要：目的观察3种神经细胞保护措施对缺氧缺血性脑损伤新生大鼠脑组织凋亡相关因子及其配体(Fas和FasL)基因表达的影响。方法选择120只7日龄Wistar大鼠,按随机数字表法将大鼠分为神经干细胞组、促红细胞生成素(EPO)组、ω-3不饱和脂肪酸组、缺氧缺血性脑损伤模型组,每组30只。神经干细胞组、EPO组、ω-3不饱和脂肪酸组于制模后经尾静脉注射5 mL神经干细胞、EPO、ω-3不饱和脂肪酸.缺氧缺血性脑损伤模型组给予等量生理盐水给药后6.12、24、48、72 h 5个时间点各组处死6只大鼠取海马组织.测定大脑海马组织Fas/FasL的基因表达,以及Toll样受体4(TLR4)、核转录因子-κB(NF-κB)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-1β、IL-6)的蛋白表达水平和细胞凋亡率。结果①mRNA表达:3个实验组给药后Fas和FasL的mRNA表达均较缺氧缺血性脑损伤模型组显著降低,以给药后24h降低最为显著,且神经干细胞组VEPO组<ω-3不饱和脂肪酸组<缺氧缺血性脑损伤模型组〔FasmRNA表达(2^-△△Ct):140.5±2.9、156.4±2.5.165.2±2.7比173.7±2.8,FasL mRNA表达(2^-△△Ct):143.1±4.3、154.6±1.5,160.7±1.4比174.7±2.8],各组间比较差异均有统计学意义(均P<0.05)。②蛋白表达:3个实验组给药后海马组织TLR4、NF-κB、TNF-α、IL-1β、IL-6的蛋白表达水平均较缺氧缺血性脑损伤模型组显著降低(TLR4/GAPDH:0.7±0.2,0.6±0.1、0.2±0.1比1.4±0.1;NF-κB/GAPDH:6.7±0.4,5.3±0.1、1.1±0.2比11.2±0.3;TNF-α/GAPDH:14.3±1.4、11.2±1.2、3.2±2.1比23.2±0.5;IL-1β/GAPDH:9.4±0.2,7.4±0.3,2.2±0.3比13.4±0.1;IL-6/GAPDH:36.2±4.4.39.3±1.5、26.2±2.1比51.4±1.4,均P<0.05).神经干细胞组上述指标的蛋白表达水平VEPO组<ω-3不饱和脂肪酸组<缺氧缺血性脑损伤模型组:③细胞凋亡率:ω-3不饱和脂肪酸组、EPO组、神经干细胞组给药后海马组织细胞凋亡率均明显低于缺氧缺血性脑损伤模型组〔(3.7±0.3)%、(3.4±0.2)%、(2.5Objective To observe the effects of 3 neuroprotective measures on the expressions of apoptosisrelated factors and their ligands(Fas and FasL)in brain tissue of neonatal rats with hypoxic ischemic brain injury.Methods One hundred and twenty Wistar rats 7 days old were selected as experimental subjects,the rats were divided into four groups:neural stem cell,erythropoietin(EPO),ω-3 unsaturated fatty acid treatment groups and hypoxic ischemic brain damage model group according to random number table method,with 30 rats in each group.Neural stem cell group,EPO group andω-3 unsaturated fatty acid group were respectively injected with neural stem cells,EPO andω-3 unsaturated fatty acid,each 5 mL via tail vein after modeling;the hypoxic ischemic brain damage model group was given equal volume of normal saline.At 6.12,24,48 and 72 hours after administration of drug,6 rats were sacrificed in each group,brain tissue was taken,the mRNA expression levels of Fas/FasL,protein expression levels of Toll-like receptor 4(TLR4),nuclear transcription factor-κB(NF-κB),tumor necrosis factor-α(TNE-α),interleukin(IL-1β,IL-6)and cell apoptotic rate in hippocampus tissue were measured.Results D mRNA expressions:the mRNA expressions of Fas and FasL of the 3 experimental groups were significantly lower than those of the hypoxic ischemic brain damage model group,the degrees of descent after administration for 24 hours were the most significant,neural stem cell treatment group<EPO treatment groupω-3 unsaturated fatty acid treatment group<hypoxic ischemic brain damage model group[Fas mRNA expression(2^-△△Ct):140.5±2.9,156.4±2.5,165.2±2.7 vs.173.7±2.&FasL mRNA expression(2^-△△Ct):143.1±4.3,154.6±1.5,160.7±1.4 vs.174.7±2.8],the differences were statistically significant(all P<0.05).(2)Protein expressions:the protein expressions of TLR4,NF-κB,TNF-α,IL-1β,IL-6 of the 3 experimental groups were significantly lower than those of the hypoxic ischemic hrain damage model group(TLR4/GAPDH:0.7±0.2,0.6±0.1,0.2±0.1 vs.1.4±0.1

关 键 词：神经干细胞 促红细胞生成素 Ω-3不饱和脂肪酸 缺氧缺血性脑损伤 新生大鼠 脑组织凋亡相关因子及其配体表达 

分 类 号：R644[医药卫生—外科学]