褪黑素治疗对创伤性脑损伤大鼠海马组织炎症反应的影响  

Effect of melatonin on inflammatory response of hippocampus in rats with traumatic brain injury

作　　者：秦荣 张珑[1] 熊鹏举 张先斌 刘宁[1] 金健辉 邓磊[1] QIN Rong;ZHANG Long;XIONG Peng-ju;ZHANG Xian-bin;LIU Ning;JIN Jian-hui;DENG Lei(The No.908th Hospital of PLA,Branch ofYingtan,None hang 330002,Jiangxi,China)

机构地区：[1]中国人民解放军联勤保障部队第908医院鹰潭院区,江西南昌330002

出　　处：《生物医学工程与临床》2021年第5期533-538,共6页Biomedical Engineering and Clinical Medicine

基　　金：中国人民解放军联勤保障部队第908医院专项课题(YGKTLC2016002)。

摘　　要：目的观察褪黑素(MEL)治疗对创伤性脑损伤(TBI)大鼠认知功能及海马组织Toll样受体4(TLR4)介导的炎性因子表达水平的影响。方法选择72只雄性SD大鼠,鼠龄3~4个月,体质量约230~280 g。按随机数字表法分为假手术组(SHAM组)、TBI组、MEL治疗组(MEL组),每组24只。采用Feency重物自由落体撞击法建立TBI模型;SHAM组大鼠开骨窗后不用撞针撞击,骨蜡封闭骨窗后直接缝合皮肤;MEL组大鼠在TBI建模后每天进行MEL(10 mg/kg)腹腔注射,每天1次,共计14次。在建模1周后,利用Morris水迷宫测试系统评估大鼠认知功能;Western blot法检测大鼠海马组织中Toll样受体4(TLR4)蛋白、核因子κB(NF-κB)P65蛋白的表达水平;酶联免疫吸附分析(ELISA)法检测大鼠海马组织中白细胞介素(IL)-1β、肿瘤坏死因子α(TNF-α)、IL-6等炎性因子的表达水平。结果Morris水迷宫检测发现,在第1天至第5天的定位航行实验中,TBI组、MEL组大鼠逃逸潜伏期较SHAM组延长(P<0.05);与TBI组比较,MEL组大鼠逃逸潜伏期缩短(P<0.05)。第6天空间探索实验发现,TBI组、MEL组大鼠穿越平台次数较SHAM组减少(P<0.05);MEL组大鼠穿越平台次数较TBI组增加(P<0.05)。Western blot检测结果发现,TBI组、MEL组大鼠海马组织中TLR4蛋白相对表达水平分别为(3.31±0.06)、(1.98±0.05),NF-κB P65蛋白相对表达水平分别为(2.96±0.07)、(2.01±0.06),均较SHAM组升高(P<0.05);MEL组大鼠海马组织中TLR4蛋白、NF-κB P65蛋白表达水平较TBI组下降(P<0.05)。ELISA检测结果发现,TBI组、MEL组大鼠海马组织中IL-1β表达水平分别为(87.65±7.26)pg/mL、(69.36±5.27)pg/mL,TNF-α表达水平分别为(56.21±3.84)pg/mL、(37.56±3.47)pg/mL,IL-6表达水平分别为(146.61±11.31)pg/mL、(97.25±12.79)pg/mL,均较SHAM组升高(P<0.05);MEL组大鼠海马组织中IL-1β、TNF-α、IL-6表达水平较TBI组下降(P<0.05)。结论大鼠在TBI后出现不同程度的认知功能障碍;MEL能够改善TBI后认知功能�Objective To observe the effects of melatonin(MEL)on cognitive function and expression of Toll-like receptor 4(TLR4)-mediated inflammatory factors in hippocampus of rats with traumatic brain injury(TBI).Methods Seventy-two male SD rats were selected,which aged 3-4 months,body mass was 230-280 g.The rats were divided into sham operation group(SHAM group),TBI group and MEL treatment group(MEL group),with 24 rats in each group.The TBI model was established by Feency method for free fall impact.In SHAM group,the bone window was closed with bone wax and skin was sutured directly.After TBI modeling,rats in MEL group were injected with MEL(10 mg/kg)intraperitoneally once a day for 14 days.One week after modeling,the cognitive function of rats was evaluated by Morris water maze.The expression levels of TLR4 protein and nuclear factor kappa-B(NF-κB)P65 protein in hippocampus were detected by Western blot,and inflammatory factor expression levels of interleukin 1β(IL-1β),tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6)were detected by enzyme linked immunosorbent assay(ELISA).Results Morris water maze test results showed that,in navigation experiment from day 1 to day 5,the escape latency of TBI group and MEL group were longer than that of SHAM group(P<0.05).Compared with TBI group,escape latency of MEL group was shorter(P<0.05).On day 6 of space exploration experiment,the crossing platform times in TBI group and MEL group were less than that in SHAM group(P<0.05);the crossing platform times in MEL group were more than that in TBI group(P<0.05);Western blot analysis results showed that the relative expression levels of TLR4 in TBI and MEL groups were(3.31±0.06),(1.98±0.05)and relative expression level of NF-κB P65 were(2.96±0.07),(2.01±0.06),which were higher than those in SHAM group(P<0.05).The relative expression levels of TLR4 and NF-κB P65 of hippocampus in MEL group were lower than those in TBI group(P<0.05).ELISA test results showed that expression level of IL-1βof hippocampus in TBI group and MEL gro

关 键 词：褪黑素 创伤性脑损伤 认知功能 TOLL样受体4 NF-κB P65 炎性因子 

分 类 号：R651.15[医药卫生—外科学]