针刺调控NLRC5抑制NF-κB通路减轻SAMP8小鼠脑内神经炎症反应的研究

Acupuncture regulation of NLRC5 inhibits NF-κB pathway and alleviates neuroinflammatory response in SAMP8 brain

关键词:  阿尔茨海默病  针刺  炎症  NLRC5  核转录因子-κB

Key words:  Alzheimer disease  acupuncture  inflammation  NLRC5  NF-κB

摘要:

[目的] 观察针刺调控NLRC5抑制核转录因子-κB（NF-κB）通路减轻SAMP8脑内神经炎症反应的作用。[方法] 将7月龄雄性快速老化鼠SAMP8及其同源正常对照小鼠SAMR1分为SAMR1组、SAMP8组和SAMP8针刺组。针刺组予以针刺治疗，采用"调神益智"针法进行治疗，取穴为水沟、内关和三阴交；针刺每日1次，持续30 d。采用Morris水迷宫检测认知能力；逆转录-聚合酶链反应（RT-PCR）和Western blot法检测海马内p65、NF-κB抑制蛋白激酶（IκK）、NLRC5 mRNA及蛋白表达；酶联免疫吸附（ELISA）法检测海马内白介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）水平。。[结果] 与SAMR1组比较，SAMP8小鼠学习记忆功能明显受损，表现为在隐蔽及反向平台实验中逃避潜伏期较长（P<0.05）；搜索平台所用策略较为僵化；在空间探索实验中，首次穿越原平台时间明显较长，穿越原平台次数较少，中环区域停留时间百分比较小（P<0.05）。脑内p-IκK、p-p65 mRNA及蛋白表达上升，而NLRC5 mRNA及蛋白表达下降；海马内IL-1β、IL-6、TNF-α水平显著增高（P<0.01）。针刺治疗后，针刺组在隐蔽平台及反向平台实验中逃避潜伏期较SAMP8组明显缩短（P<0.05），且在空间探索实验中，首次穿越原平台时间、穿越原平台次数、中环区域停留时间百分比均明显改善（P<0.05）。针刺能明显下调SAMP8脑内p-IκK、p-p65 mRNA及蛋白表达，而提高NLRC5 mRNA及蛋白表达。针刺能够降低SAMP8海马内上述炎症因子水平，特别是IL-6降低明显。。[结论] SAMP8鼠脑内存在炎症反应激活现象，针刺通过提高NLRC5来抑制NF-κB活化，减少炎症因子释放，降低脑内炎症反应，从而改善神经元存活微环境，进而改善SAMP8小鼠的认知障碍。

Abstract:

[Objective] To investigate the effect of acupuncture on NLRC5 inhibition of NF-κB pathway in alleviating neuroinflammatory response in SAMP8 brain.[Methods] Seven-month-old male rapidly aging mouse SAMP8 and its homologous normal control mouse SAMR1 were divided into SAMR1 group,SAMP8 group and SAMP8 acupuncture group. The acupuncture group was treated with "Tiaoshen Yizhi" acupuncture method once daily for 30 days and the acupoints included Renzhong,Neiguan and Sanyinjiao. The cognitive ability was detected by Morris water maze. The expressions of p65,IκK and NLRC5 mRNA and protein in hippocampus were detected by RT-PCR and Western blot. The levels of IL-1β,IL-6 and TNF-α in hippocampus were detected by ELISA.[Results] Compared with the SAMR1 group,the learning and memory function of SAMP8 mice was significantly impaired,which showed that the escape latency was longer in concealed and reverse platform experiments (P<0.05),and the strategy used in the search platform was more rigid. In the space exploration experiment,the time to cross the original platform for the first time was significantly longer,the number of crossing the original platform was less,and the percentage of staying time in the central area was smaller (P<0.05). The expression of p-IκK and p-p65 mRNA and protein increased,while the expression of NLRC5 mRNA and protein decreased. The levels of IL-1β,IL-6 and TNF-α in hippocampus were significantly increased (P<0.01). After acupuncture treatment,the escape latency of the acupuncture group in the concealed platform and reverse platform experiment was significantly shorter than that of the SAMP8 group (P<0.05),and in the space exploration experiment,the time of crossing the original platform for the first time,the number of crossing the original platform,and the central region The percentage of residence time was significantly improved (P<0.05). Acupuncture can significantly down-regulate the expression of p-IκK and p-p65 mRNA and protein in SAMP8 brain,and increase the expression of NLRC5 mRNA and protein. Acupuncture can reduce the level of inflammatory factors in the hippocampus of SAMP8,especially IL-6.[Conclusions] There is an activation of inflammatory response in the brain of SAMP8 mice. Acupuncture inhibits the activation of NF-κB by increasing NLRC5,reduces the release of inflammatory factors,reduces the inflammatory response in the brain,thereby improving the survival microenvironment of neurons,and improving the recognition of SAMP8 mice.