早期抑制TLR4对新生大鼠HIBD后青春期脑海马免疫功能的影响  

Effect of early inhibition of TLR4 on hippocampal immune function to adolescence after neonatal HIBD

作　　者：黄晓丽 欧阳志翠 吴祥宏 李燕[2] 黄赟 刘国琼 陆诗微 唐振 Huang Xiaoli;Ouyang Zhicui;Wu Xianghong;Li Yan;Huang Yun;Liu Guoqiong;Lu Shiwei;Tang Zhen(Dept of Neonatology,Affiliated Hospital of Guilin Medical University,Guilin 541001;Class of 2019 Clinical Medicine Pediatrics,Guilin Medical University,Guilin 541001)

机构地区：[1]桂林医学院附属医院新生儿科,桂林541001 [2]桂林医学院,桂林541001

出　　处：《安徽医科大学学报》2023年第8期1317-1322,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82160301);桂林市科技局创新平台和人才计划项目(编号:20210218-5);2021年广西区级大学生创新创业训练计划项目(编号:S202110601051)。

摘　　要：目的探讨Toll样受体4(TLR4)早期抑制在调节新生大鼠缺氧缺血性脑损伤(HIBD)后青春期脑海马免疫功能中的作用。方法生后7 d新生大鼠随机分为Control组、缺氧缺血(HI)组及HI^(+)TLR4抑制剂TAK-242组(TAK-242组)。HI后3 d免疫组化检测大鼠脑海马TLR4表达;HI后21 d免疫荧光检测大鼠脑海马CA1区Iba-1^(+)、GFAP^(+)、CD161^(+)、MPO^(+)及CD3^(+)细胞的数量变化;免疫组化检测大鼠脑海马CA1区黏附因子ICAM-1、补体C3a表达;Western blot检测大鼠脑海马IL-1β、TNF-α、IL-10表达。结果HI后3 d大鼠脑海马CA1、CA3及DG区TLR4表达,HI组较Control组升高(P<0.01或P<0.05),而TAK-242组较HI组降低(P<0.05)。HI后21 d大鼠脑海马CA1区GFAP^(+)细胞数量,TAK-242组较HI组降低(P<0.05);CD3^(+)T淋巴细胞数量,HI组较Control组增加(P<0.05),而TAK-242组较HI组差异无统计学意义;HI后21 d各组大鼠脑海马CA1区Iba-1^(+)、MPO^(+)、CD161^(+)细胞数量,ICAM-1、C3a因子表达及海马组织TNF-α、IL-1β及IL-10表达差异无统计学意义。结论早期抑制TLR4可通过减轻新生儿HIBD后脑海马星形胶质细胞增加而改善其青春期神经免疫紊乱。Objective To investigate the role of early inhibition of Toll-like receptor 4(TLR4)in regulating hippocampal neuroimmune responseto adolescent ratsafter neonatal hypoxic-ischemic brain damage(HIBD).Methods Postnatal day 7 rats were randomized into controlgroup,hypoxic ischemia(HI)group,and HI+TAK-242(the specific inhibitor of TLR4)(TAK-242)group.The expression of TLR4 in rat hippocampus was detected by immuno-histochemistry at 3 days after HI.Immunofluorescence were used to determine the number of Iba-1^(+),GFAP^(+),CD161^(+),MPO+and CD3^(+)cells in the hippocampus at 21 days after HI.Immunohistochemistry was used to detect ICAM-1 and C3 a expression in the hippocampal CA1 region;and Western blot was used to detect tumor necrosis factor interleukin IL-1β,TNF-α and IL-10 expression.Results Compared with control group,significantly raised TLR4 expression was observed in the left hippocampal CA1,CA3 and DG regions(P0.01 or P0.05),while the expression in the TAK-242 group lowered compared to the HI group(P0.05).The number of GFAP+cells in the CA1 area of the hippocampus in the TAK-242 group of neonatal rats decreased compared to which in the HI group at 21 days after HI(P0.05),but the number of CD3^(+)T lymphocytes in the hippocampal CA1 area of new born rats in the HI group increased compared to which in the Control group(P0.05),but the difference between TAK-242 and the Control group was not statistically significant.The number of Iba-1^(+)cells,MPO^(+)cells,CD161^(+)cells,the expression of ICAM-1 and C3a in hippocampal CA1 region,and the expression of TNF-α,IL-1β and IL-10 in hippocampus of rats were not different among groups at 21 days after HIBD.Conclusion Early inhibition of TLR4 may ameliorate adolescent neuroimmune disorders by reducing the increase of hippocampal astrocytesafter neonatal HIBD.

关 键 词：缺氧缺血性脑损伤 海马 TLR4 大鼠 神经免疫 

分 类 号：R722.1[医药卫生—儿科]