p62/NF-κB通路调控HIV-1 gp120 V3环所致小鼠神经炎症的机制研究  

Mechanism of p62/NF-κB pathway regulating HIV-1 gp120 V3 loop-in⁃duced neuroinflammation in mice

作　　者：何翰阳 颜学勤 温赛娴 干李梦 王琳琳[1] 王会丽 唐海杰 潘锐[2] 付咏梅[1] 董军[1] HE Han-yang;YAN Xue-qin;WEN Sai-xian;GAN Li-meng;WANG Lin-lin;WANG Hui-li;TANG Hai-jie;PAN Rui;FU Yong-mei;DONG Jun(Department of Pathophysiology,School of Medicine,Laboratory of Pathophysiology,National Administration of Tradi-tional Chinese Medicine,GHM Institute of CNS Regeneration,Jinan University,Guangzhou 510632,China;Department of Orthopaedics,The First Affiliated Hospital of Jinan University,Guangzhou 510632,China)

机构地区：[1]暨南大学基础医学与公共卫生学院病理生理学系,国家中医药管理局病理生理科研实验室,粤港澳中枢神经再生研究院,广东广州510632 [2]暨南大学附属第一医院骨科,广东广州510632

出　　处：《中国病理生理杂志》2022年第6期978-985,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81974185);广东省自然科学基金资助项目(No.2019A1515012024,No.2022A1515010268);高等学校学科创新引智计划项目(No.B14036)。

摘　　要：目的:探讨自噬关键蛋白p62在HIV-1 gp120 V3环所致小鼠神经炎症中的作用及相关信号分子机制。方法:野生型C57BL6小鼠随机分成4组:空白组、假手术组(人工脑脊液组)、模型组(gp120 V3环组)及gp120 V3环+NF-κB活化阻滞剂BAY 11-7082组,每组12只。用Morris水迷宫检测小鼠学习记忆能力;免疫荧光染色检测海马和皮层Iba-1表达水平;ELISA法检测海马和皮层炎症因子的表达水平;Western blot检测海马和皮层相关蛋白表达水平。结果:(1)Morris水迷宫结果显示,与空白组相比,模型组小鼠逃避潜伏期显著延长(P<0.01),平台区域停留时间及穿越平台次数显著减少(P<0.05);与模型组相比,gp120 V3环+BAY 11-7082组小鼠逃避潜伏期显著缩短(P<0.01),平台区域停留时间及穿越平台次数显著增加(P<0.05)。(2)免疫荧光染色结果显示,与空白组相比,模型组海马和皮层Iba-1荧光强度显著增强(P<0.05);与模型组相比,gp120 V3环+BAY 11-7082组海马和皮层Iba-1荧光强度显著降低(P<0.05)。(3)ELISA结果显示,与空白组相比,模型组IL-1β、TNF-α和IL-6水平均显著上调(P<0.05);与模型组相比,gp120 V3环+BAY 11-7082组IL-1β、TNF-α和IL-6水平显著下调(P<0.05)。(4)Western blot结果显示,与空白组相比,模型组p62蛋白表达水平显著上调(P<0.01),p-p65/p65及p-IκB/IκB比值均显著升高(P<0.01);与模型组相比,gp120 V3环+BAY 11-7082组p62蛋白显著下调(P<0.05),p-p65/p65及p-IκB/IκB比值均显著下降(分别P<0.01和P<0.05)。结论:HIV-1 gp120 V3环所致小鼠学习记忆功能障碍的机制可能与通过激活p62/NF-κB信号通路引起神经炎症有关,p62-NF-κB正反馈环的抑制可能会减轻神经炎症。AIM:To explore the role of autophagy key protein p62 in HIV-1 gp120 V3 loop-induced neuroinflammation in mice and its molecular mechanisms.METHODS:Wild-type C57BL6 mice were randomly divided into 4 groups:blank group,sham group(artifical cerebrospinal fluid group),model group(gp120 V3 loop group)and gp120 V3 loop+NF-κB activation blocker BAY 11-7082 group,with 12 mice in each group.Learning and memory ability of the mice was detected by Morris water maze.The Iba-1 expression in the hippocampus and cortex was detected by immunofluorescence.The expression levels of inflammatory factors in the hippocampus and cortex were examined by ELISA.Protein ex⁃pression levels in the hippocampus and cortex were examined by Western blot.RESULTS:The results of Morris water maze showed that compared with blank group,the escape latency of the mice in model group was significantly prolonged(P<0.01),and the residence time in the target platform and the number of crossing the platform were significantly reduced(P<0.05).Compared with model group,the escape latency of gp120 V3 loop+BAY 11-7082 group was significantly shortened,and the residence time in the target platform and the number of crossing the platform were significantly increased(P<0.05).Immunofluorescence results showed that compared with blank group,the fluorescence intensity of Iba-1 in the hippocampus and cortex of the mice in model group was significantly up-regulated(P<0.05),while that in gp120 V3 loop+BAY 11-7082 group was significantly down-regulated compared with model group(P<0.05).ELISA results showed that compared with blank group,the levels of IL-1β,TNF-αand IL-6 in model group were significantly increased(P<0.05),while those in gp120 V3 loop+BAY 11-7082 group were significantly lowered compared with model group(P<0.01).Western blot showed that compared with blank group,the expression of p62 protein was significantly up-regulated(P<0.01),and the NF-κB signaling pathway was activated in model group,indicated by the increases in the ratios of p-IκB/IκB and p

关 键 词：HIV相关神经认知障碍 HIV-1 gp120 V3环 p62/NF-κB信号通路 神经炎症 

分 类 号：R363[医药卫生—病理学] R741[医药卫生—基础医学]