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肠道菌群失调驱动的辅助性 T 细胞 17 细胞激活介导麻醉/手术引起的老年大鼠围手术期神经认知障碍。

Intestinal dysbacteriosis-propelled T helper 17 cells activation mediate the perioperative neurocognitive disorder induced by anesthesia/surgery in aged rats.

机构信息

Department of Anesthesiology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, Jiangsu 210009, China; Department of Anesthesiology, Women's Hospital of Nanjing Medical University (Nanjing Maternity and Child Health Care Hospital), Nanjing, Jiangsu 210004, China.

Department of Anesthesiology, Women's Hospital of Nanjing Medical University (Nanjing Maternity and Child Health Care Hospital), Nanjing, Jiangsu 210004, China.

出版信息

Neurosci Lett. 2022 Jul 13;783:136741. doi: 10.1016/j.neulet.2022.136741. Epub 2022 Jun 15.

Abstract

BACKGROUND

Perioperative neurocognitive disorders (PND) is a common postoperative disease in elderly patients, but its pathogenesis remains unclear.

METHODS

Exploratory laparotomy was performed to establish PND model under sevoflurane anesthesia. 16S rRNA high-throughput sequencing was used to detect the changes of intestinal flora. Antibiotics were used to relatively eliminate intestinal flora before anesthesia/surgery, and behavior tests, such as open field, Y maze, and fear conditioning tests were applied to detect the changes of memory ability. The number of Th17 cells and Foxp3 cells was detected by flow cytometry in the Peyer's patches (PP), mesenteric lymph nodes (MLN), blood and brain. Western blot was used to detect the expression of IL17, IL17RA, IL6 and IL10 in the hippocampus. Immunofluorescence was used to detect the expression of IL17, IL17R and IBA1 (ionized calcium binding adaptor molecule1) in the hippocampus.

RESULTS

Anesthesia/surgery caused intestinal flora imbalance and induced neurocognitive impairment, increased the number of Th17 cells in the PP, MLN, blood and brain, increased the level of IL17, IL17R and inflammatory factors production in the hippocampus. Antibiotics administration before anesthesia/surgery significantly decreased the number of Th17 cells and the level of IL17, IL17R and inflammatory factors production, and improved the memory function. In addition, we found that IL17R was co-labeled with IBA1 in a large amount in the hippocampus through immunofluorescence double-staining.

CONCLUSION

Our study suggested that intestinal dysbacteriosis-propelled T helper 17 cells activation and IL17 secretion might play an important role in the pathogenesis of PND induced by anesthesia/surgery in aged rats.

摘要

背景

围手术期神经认知障碍(PND)是老年患者术后常见的疾病,但发病机制尚不清楚。

方法

采用七氟醚麻醉下剖腹探查建立 PND 模型。采用 16S rRNA 高通量测序检测肠道菌群变化。麻醉/手术前应用抗生素相对消除肠道菌群,应用旷场、Y 迷宫、恐惧条件反射等行为学检测记忆能力变化。流式细胞术检测派尔集合淋巴结(PP)、肠系膜淋巴结(MLN)、血液和大脑中 Th17 细胞和 Foxp3 细胞的数量。Western blot 检测海马中 IL17、IL17RA、IL6 和 IL10 的表达。免疫荧光法检测海马中 IL17、IL17R 和 IBA1(离子钙结合衔接分子 1)的表达。

结果

麻醉/手术导致肠道菌群失衡,引起神经认知障碍,增加 PP、MLN、血液和大脑中 Th17 细胞数量,增加海马中 IL17、IL17R 和炎症因子产生水平。麻醉/手术前应用抗生素可显著减少 Th17 细胞数量及 IL17、IL17R 和炎症因子产生水平,并改善记忆功能。此外,我们通过免疫荧光双重染色发现,IL17R 在海马中大量与 IBA1 共标记。

结论

本研究表明,肠道菌群失调驱动的 Th17 细胞激活和 IL17 分泌可能在老年大鼠麻醉/手术引起的 PND 发病机制中起重要作用。

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