TRIM31在七氟醚麻醉致老龄大鼠认知功能障碍中的作用-现代医学

TRIM31在七氟醚麻醉致老龄大鼠认知功能障碍中的作用

单位：利川市人民医院麻醉科, 湖北利川 445400

分类号：R749

出版年·卷·期（页码）：2022·50·第十期（1283-1289）

摘要：

目的：探究三结构域蛋白31（TRIM31）在七氟醚麻醉致老龄大鼠认知功能障碍中的作用及可能的作用机制。方法：60只SD老龄大鼠随机分为对照组、七氟醚麻醉组（SEV组）、七氟醚麻醉+空载慢病毒组（SEV+Lv-NC组）和七氟醚麻醉+TRIM31过表达慢病毒组（SEV+Lv-TRIM31组），每组15只。采用3%七氟醚麻醉6 h建立麻醉模型，SEV+Lv-NC组和SEV+Lv-TRIM31组分别于麻醉前2天经侧脑室注射Lv-NC或Lv-TRIM31慢病毒溶液。麻醉结束后第2天行Morris水迷宫实验检测大鼠认知功能；随后取海马组织，苏木精-伊红染色观察海马组织病理学变化；原位末端标记染色观察海马组织细胞凋亡情况；分光光度法检测海马组织中Caspase-3活性；酶联免疫吸附测定法检测海马组织中小鼠白介素1β（IL-1β）、白介素6（IL-6）、肿瘤坏死因子α（TNF-α）和白介素18（IL-18）含量；实时荧光定量PCR检测海马组织中TRIM31 mRNA水平；蛋白质免疫印迹法检测海马组织中TRIM31、NOD样受体蛋白3（NLRP3）、衔接分子凋亡相关斑点样蛋白（ASC）和半胱氨酸蛋白酶1（Caspase-1）蛋白表达水平。结果：与对照组比较，SEV组大鼠逃避潜伏期延长，目标象限停留时间和穿越原平台次数减少，海马组织结构破坏严重，同时海马组织中细胞凋亡率、半胱氨酸蛋白酶3（Caspase-3）活性、IL-1β、IL-6、TNF-α和IL-18含量以及NLRP3、ASC和Caspase-1蛋白表达水平均显著上升（P<0.01）；与SEV组比较，SEV+Lv-TRIM31组大鼠逃避潜伏期缩短，目标象限停留时间和穿越原平台次数增加，海马组织结构恢复，同时海马组织中细胞凋亡率、Caspase-3活性、IL-1β、IL-6、TNF-α和IL-18含量以及NLRP3、ASC和Caspase-1蛋白表达水平均显著下降（P<0.01），SEV+Lv-NC组以上指标差异无统计学意义（P>0.05）。结论：过表达TRIM31可通过抑制NLRP3炎性小体诱导的神经炎症，改善七氟醚麻醉诱发的老龄大鼠认知功能障碍。

Objective: To explore the role and possible mechanism of tripartite motif containing 31(TRIM31) in cognitive dysfunction in aged rats induced by sevoflurane anesthesia. Methods: Sixty SD rats were randomly divided into Control group, sevoflurane anesthesia group(SEV), sevoflurane anesthesia+no-load lentivirus group (SEV+LV-NC), and sevoflurane anesthesia+TRIM31 overexpressed lentivirus group (SEV+LV-TRIM31), with 15 rats in each group. The anesthesia model was established with 3% sevoflurane for 6 h. The groupsof SEV+LV-NC and SEV+LV-TRIM31 were injected with LV-NC or LV-TRIM31 lentivirus solution through the lateral ventricle one day before anesthesia. After 2 days of anesthesia, all rats underwent Morris water maze experiment to test their cognitive function. Then all rats were sacrificed and the hippocampal tissues were taken, and the histopathological changes were observed by HE staining. Apoptosis of hippocampal cells was observed by TUNEL staining. The activity of Caspase-3 was detected by spectrophotometry. IL-1β, IL-6, TNF-α and IL-18 were detected by ELISA. TRIM31 mRNA levels were detected by qRT-PCR. The expression levels of TRIM31, NLRP3, ASC and Caspase-1 proteins in hippocampus were detected by Western blotting. Results: Compared with the Control group, the SEV group rats had longer escape incubation period, reduced residence time in the target quadrant and reduced times of crossing the original platform, and the hippocampal tissue structure was seriously damaged. Meanwhile, the apoptosis rate, the activity of Caspase-3, the level of IL-1β, IL-6, TNF-α and IL-18, the protein expression level of NLRP3, ASC and Caspase-1 in hippocampus were significantly increased(P<0.01). Compared with the SEV group, the SEV+Lv-TRIM31 group rats had shorter evasive latency, higher target quadrant stay time and more times of crossing the original platform, and the hippocampal structure was restored. Meanwhile, the apoptosis rate, Caspase-3 activity, levels of IL-1β, IL-6, TNF-α and IL-18, the protein expression levels of NLRP3, ASC and Caspase-1 in hippocampus were significantly decreased(P<0.01). However, there was no statistical significance in the SEV+LV-NC group(P>0.05). Conclusion: Overexpression of TRIM31 can improve cognitive dysfunction induced by sevoflurane in aged rats by inhibiting neuro inflammation induced by NLRP3 inflammasome.

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