Necrostatin-1预先给药对膝关节炎慢性痛老龄大鼠术后认知功能的影响  

Effect of necrostatin-1 pre-treatment on postoperative cognitive function in aged rats with chronic pain due to knee arthritis

作　　者：于丽丽[1] 尹春平 赵娟[3] 宋盼盼 李春雷 孙秀伟[1] 李晨晨 常玉林[1] Yu Lili;Yin Chunping;Zhao Juan;Song Panpan;Li Chunlei;Sun Xiuwei;Li Chenchen;Chang Yulin(Department of Anesthesiology,Cangzhou Central Hospital,Cangzhou 061000,China;Department of Anesthesiology,The First Hospital of Hebei Medical University,Shijiazhuang 050030,China;Experimental Centre for Teaching of Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]沧州市中心医院麻醉科,沧州061000 [2]河北医科大学第一医院麻醉科,石家庄050030 [3]河北医科大学教学实验中心,石家庄050011

出　　处：《中华麻醉学杂志》2023年第11期1329-1334,共6页Chinese Journal of Anesthesiology

基　　金：2022年度河北省医学科学研究课题计划(20220339)。

摘　　要：目的评价Necrostatin-1预先给药对膝关节炎慢性痛老龄大鼠术后认知功能的影响。方法健康雄性SD大鼠120只, 22月龄, 体质量550 ~ 600 g, 采用随机数字表法分为4组(n=30):膝关节炎慢性痛组(P组)、膝关节炎慢性痛+手术组(PS组)、二甲基亚砜(DMSO)+膝关节炎慢性痛+手术组(DMSO+PS组)和Necrostatin-1 +膝关节炎慢性痛+手术组(Nec-1+PS组)。4组均于左膝关节腔内注射碘乙酸盐(MIA)1 mg制备膝关节炎模型。PS组、DMSO+PS组和Nec-1+PS组于注射MIA后12周时行七氟烷麻醉下剖腹探查术。Nec-1+PS组和DMSO+PS组分别于术前1 h腹腔注射Necrostatin-1 6.25 mg/kg或等剂量DMSO。于术后7 d时采用Morris水迷宫实验评估认知功能, 采用免疫荧光染色法确定海马区小胶质细胞活化率;取海马组织, 采用流式细胞术测定神经元坏死率, 采用Western blot法测定受体相互作用蛋白激酶1(RIPK1)和磷酸化混合谱系激酶结构域样蛋白(p-MLKL)的表达, 采用ELISA法测定TNF-α、IL-1β和IL-6的含量。结果与P组比较, PS组、DMSO+PS组和Nec-1+PS组逃避潜伏期延长, 原平台象限停留时间缩短, 穿越原平台位置次数减少, 小胶质细胞活化率和海马神经元坏死率增加, 海马组织RIPK1和p-MLKL表达上调, TNF-α、IL-1β和IL-6含量升高(P<0.05);与PS组和DMSO+PS组比较, Nec-1+PS组逃避潜伏期缩短, 原平台象限停留时间延长, 穿越原平台位置次数增加, 小胶质细胞活化率和海马神经元坏死率降低, 海马组织RIPK1和p-MLKL表达下调, TNF-α、IL-1β和IL-6含量降低(P<0.05)。结论 Necrostatin-1预先给药可改善膝关节炎慢性痛老龄大鼠术后认知功能, 其机制可能与抑制海马神经元程序性坏死, 减轻神经炎症有关。Objective To evaluate the effect of necrostatin-1(Nec-1)pre-treatment on postoperative cognitive function in aged rats with chronic pain due to knee arthritis.Methods One hundred and twenty healthy male Sprague-Dawley rats,aged 22 months,weighing 550-600 g,were divided into 4 groups(n=30 each)using a random number table method:chronic pain due to knee arthritis group(group P),chronic pain due to knee arthritis+operation group(group PS),dimethyl sulfoxide(DMSO)+chronic pain due to knee arthritis+operation group(DMSO+PS group),and necrostatin-1+chronic pain due to knee arthritis+operation group(Nec-1+PS group).The inflammation-induced knee arthritis model was developed by injecting monosodium iodoacetate(MIA)into the left joint cavity.The exploratory laparotomy under sevoflurane anesthesia was performed at 12 weeks after intra-articular MIA injection.In Nec-1+PS group and DMSO+PS group,necrosstatin-16.25 mg/kg and the equal dose of DMSO were intraperitoneally injected at 1 h before surgery,respectively.At 7 days after surgery,the Morris water maze test was used to evaluate the cognitive function,the activation of microglial cells in the dentate gyrus of hippocampus was observed by immunofluorescent staining,and the activation rate of microglia cells was calculated,the necrosis rate of neurons was determined by flow cytometry,the expression of receptor-interacting protein kinase 1(RIPK1)and phosphorylated mixed lineage kinase domain-like protein(p-MLKL)was determined by Western blot,and the contents of tumor necrosis factor-alpha(TNF-α),interleukin-1beta(IL-1β)and IL-6 were determined by enzyme-linked immunosorbent assay.Results Compared with P group,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,and the number of crossing the original platform was reduced,the activation rate of microglia cells in the hippocampal dentate gyrus and necrosis rate of hippocampal neurons were increased,the expression of RIPK1 and p-MLKL was up-regulated,and the conten

关 键 词：骨关节炎 膝 老年人 术后认知并发症 Necrostatin-1 

分 类 号：R965[医药卫生—药理学]