IL-6反式信号通路在小鼠围术期神经认知障碍中的作用  

Role of IL-6 trans-signaling pathway in perioperative neurocognitive disorder in mice

作　　者：张宇 孙美艳 胡军[1] 张野[1] Zhang Yu;Sun Meiyan;Hu Jun;Zhang Ye(Department of Anesthesiology and Perioperative Medicine,The Second Hospital of Anhui Medical University,Key Laboratory of Anesthesiology and Perioperative Medicine of Anhui Higher Education Institutes,Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医科大学第二附属医院麻醉与围术期医学科,麻醉与围术期医学安徽普通高校重点实验室,合肥230601

出　　处：《中华麻醉学杂志》2022年第3期274-278,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81970231,81901086)。

摘　　要：目的探讨IL-6反式信号通路在小鼠围术期神经认知障碍中的作用。方法SPF级健康雄性C57BL/6野生型小鼠和IL-6R^(-/-)小鼠各84只,12~14周龄,体重25~35 g。采用随机数字表法将野生型小鼠分为4组(n=21):假手术组(SH组)、手术组(S组)、sgp130Fc(特异性IL-6反式信号通路阻断剂)组(F组)和sgp130Fc+手术组(FS组)。S组和FS组在七氟烷麻醉下行胫骨骨折复位内固定术,SH组和F组行七氟烷麻醉但不施行手术。麻醉前FS组和F组腹腔注射sgp130Fc 10 mg/kg。术后24 h时腹腔静脉采血,采用ELISA法测定血浆IL-6、IL-1β和TNF-α浓度,随后处死小鼠取海马,采用ELISA法测定海马IL-6、IL-1β和TNF-α含量,免疫荧光染色法观察海马DG区小胶质细胞的激活情况(n=6);术后第3天进行场景性恐惧逃避实验测试(n=15)。采用随机数字表法将IL-6R^(-/-)小鼠分为4组(n=21):假手术组(KO-SH组)、手术组(KO-S组)、saline组(KO-C组)和特异性IL-6反式信号通路激活剂hyper IL-6组(KO-H组)。KO-SH组和KO-S组的处理措施分别同SH组和S组。KO-C组腹腔注射100μl 0.9%NaCl溶液,KO-H组腹腔注射100μl hyper IL-640μg/kg,24 h后腹腔静脉采血,采用ELISA法测定血浆IL-6、IL-1β和TNF-α浓度,随后处死小鼠取海马,采用ELISA法测定海马IL-6、IL-1β和TNF-α含量,免疫荧光染色法观察海马DG区小胶质细胞活化水平(n=6);术后第3天进行场景性恐惧逃避实验测试(n=15)。结果与SH组比较,S组小鼠场景恐惧测试僵直时间百分比降低,海马小胶质细胞活化水平、血浆和海马IL-6、IL-1β和TNF-α水平升高(P<0.05);与S组比较,FS组小鼠场景恐惧测试僵直时间百分比增加,海马小胶质细胞活化水平、血浆和海马IL-6、IL-1β和TNF-α水平降低(P<0.05)。KO-S组和KO-SH组小鼠场景恐惧测试僵直时间百分比、海马小胶质细胞活化水平、血浆和海马IL-6、IL-1β和TNF-α水平差异无统计学意义(P>0.05);与KO-C组比较,KO-H组小鼠场景恐惧测�Objective To investigate the role of IL-6 trans-signaling pathway in perioperative neurocognitive disorder in mice.Methods Eighty-four SPF healthy male C57BL/6 wild-type mice and 84 SPF healthy male IL-6R^(-/-)mice,aged 12-14 weeks,weighing 25-35 g,were used.The 84 wild-type mice were divided into 4 groups(n=21 each)using a random number table method:sham group(SH group),surgery group(S group),sgp130Fc(specific IL-6 trans-signaling pathway blocker)group(F group),and sgp130Fc+surgery group(FS group).In S group and FS group,internal fixation was performed under general anesthesia with sevoflurane after tibial fracture.Mice only received anaesthesia with sevoflurane in SH group and F group.In FS group and F group,sgp130Fc 10 mg/kg was intraperitoneally injected before anesthesia.Blood samples were collected from the celiac vein at 24 h after surgery for determination of the concentrations of interleukin 6(IL-6),IL-1βand tumor necrosis factor(TNF)-αin plasma by enzyme-linked immunosorbent assay(ELISA).Then the mice were sacrificed,brains were removed,and hippocampal tissues were obtained for measurement of the contents of IL-6,IL-1βand TNF-α(by ELISA)and for observation of activation of microglias in the hippocampal DG region(by immunofluorescence staining,n=6).Cognitive function was evaluated by contextual fear conditioning test(n=15)on 3 days after surgery.Eighty-four IL-6R^(-/-)mice were randomly divided into 4 groups(n=21 each):sham group(KO-SH group),surgery group(KO-S group),saline group(KO-C group),and hyper IL-6(specific IL-6 trans-signaling pathway activator)group(KO-H group).The treatment in KO-SH group and KO-S group was the same as those previously described in SH group and S group,respectively.0.9%NaCl solution 100μl was intraperitoneally injected in KO-C group,100μl hyper IL-640μg/kg was intraperitoneally injected in KO-H group,and 24 h later blood was collected from the celiac vein for measurement of the concentrations of IL-6,IL-1βand TNF-αin plasma by ELISA.Then the mice were sacrificed,brai

关 键 词：白细胞介素6 神经认知障碍 围手术期 

分 类 号：R619[医药卫生—外科学]