国际麻醉学与复苏杂志   2024, Issue (3): 2-2
    
MCC950抑制NLRP3/caspase‑1通路改善体外循环大鼠围手术期神经认知障碍的作用研究
丁国鹏, 付益计, 孙莹杰, 刁玉刚, 宋丹丹1()
1.锦州医科大学北部战区总医院研究生培养基地
MCC950 improves perioperative cognitive disorders in rats undergoing cardiopulmonary bypass through inhibiting the NOD like receptor thermal protein domain associated protein 3/caspase‑1 pathway
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摘要：

目的 研究NOD样受体热蛋白结构域相关蛋白3（NLRP3）炎症小体抑制剂MCC950对体外循环（CPB）大鼠围手术期神经认知障碍（PND）的保护作用。 方法 清洁级成年雄性SD大鼠30只，体重350~450 g，采用随机数字表法分为3组（每组10只）：假手术组（S组）、CPB组（C组）和CPB+MCC950（CM组）。S组不建模，C组和CM组建立大鼠CPB模型。CM组在CPB前24、1 h和术后24 h注射MCC950 10 mg/kg，S组和C组注射等容量生理盐水。术后3 d大鼠进行水迷宫测试，记录逃避潜伏期和穿越平台次数。随后处死大鼠，取海马组织，采用酶联免疫吸附测定法检测白细胞介素（IL）‑18和IL‑1β含量，分光光度法检测胱天蛋白酶（caspase）‑1活性，免疫印迹法（Western‑blot）检测NLRP3、凋亡相关斑点样衔接蛋白（ASC）、胱天蛋白酶原（pro‑caspase‑1）和消皮素D（GSDMD）蛋白含量，实时荧光定量聚合酶链反应（FQ-PCR）检测NLRP3、ASC、caspase‑1 mRNA表达水平。 结果 与S组比较，C组和CM组大鼠逃避潜伏期较长（均P<0.05），穿越平台次数较少（均P<0.05）；海马组织中IL‑18、IL‑1β含量和caspase‑1酶活性较高（均P<0.05），NLRP3、ASC、pro‑caspase‑1和GSDMD蛋白含量较高（均P<0.05），NLRP3、ASC、caspase‑1 mRNA表达较高（均P<0.05）。与C组比较，CM组大鼠逃避潜伏期较短（P<0.05），穿越平台次数较多（P<0.05）；海马组织中IL‑1β、IL‑18含量和caspase‑1酶活性较低（均P<0.05），NLRP3、ASC、pro‑caspase‑1和GSDMD蛋白含量较低（均P<0.05），NLRP3、ASC、caspase‑1 mRNA表达较低（均P<0.05）。 结论 MCC950可抑制NLRP3/caspase‑1通路改善体外循环大鼠PND。
关键词： NOD样受体热蛋白结构域相关蛋白3； 胱天蛋白酶； 围手术期神经认知障碍；
Abstract:

Objective To investigate the protective effect of MCC950, the NOD like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome inhibitor on perioperative neurocognitive disorders (PND) in rats undergoing cardiopulmonary bypass (CPB). Methods According to the random number table method, 30 clean adult male SD rats, weighing 350‒450 g, were divided into three groups (n=10): a sham operation group (group S), a CPB group (group C) and a CPB+MCC950 (group CM). Rats in group S were not subject to CPB, while CPB was modeled in group C and group CM. Furthermore, group CM was injected with MCC950 at 10 mg/kg 24 h before CPB, 1 h before CPB, and 24 h after CPB, while group S and group C were injected with the equal volume of normal saline. Then, the water maze test was carried out 3 days after the operation to record the escape latency and the number of crossing the platform. Subsequently, the rats were sacrificed, and the hippocampal tissues were collected. Interleukin (IL)‑18 and IL‑1β contents were detected by enzyme‑linked immunosorbent assay, and caspase‑1 activity was detected by spectrophotometry. The protein levels of NLRP3, apoptosis‑associated speck‑like protein (ASC), pro‑caspase‑1 and gasdermin D (GSDMD) were detected by Western blot. The mRNA levels of NLRP3, ASC, and caspase‑1 were detected by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR). Results Compared with group S, group C and group CM showed prolonged escape latency (all P<0.05), with a reduced number of crossing the platform (all P<0.05), as well as increased contents of IL‑18 and IL‑1β and enhanced activity of caspase‑1 in hippocampal tissues (all P<0.05), up‑regulated protein levels of NLRP3, ASC, pro‑caspase‑1, and GSDMD (all P<0.05), and up-regulated mRNA levels of NLRP3, ASC, and caspase‑1 (all P<0.05). Compared with group C, group CM presented shortened escape latency (all P<0.05), with an increased number of crossing the platform (P<0.05), as well as decreased contents of IL‑1β and IL‑18 and weakened activity of caspase‑1 in hippocampal tissues (all P<0.05), down-regulated protein levels of NLRP3, ASC, pro‑caspase‑1 and GSDMD (all P<0.05), and down-regulated mRNA levels of NLRP3, ASC, and caspase‑1 (all P<0.05). Conclusion MCC950 can inhibit the NLRP3/caspase‑1 pathway to improve PND in CPB rats.
Key words: NOD like receptor thermal protein domain associated protein 3; Caspase; Perioperative neurocognitive disorders; Cardiopulmonary bypass; MCC950