Department of Anesthesiology, Linyi Central Hospital, Linyi, Shandong, China.
Curr Neurovasc Res. 2020;17(1):79-85. doi: 10.2174/1567202617666200128142728.
The current study was conducted in order to investigate the role of Forkhead box O1 and p21-mediated macrophage polarization in postoperative cognitive dysfunction induced by sevoflurane.
There involved a total of 30 healthy mice that were randomly divided into two groups: control group (without any treatment) and anaesthesia group (treated with sevoflurane inhalation). The effects of sevoflurane on cognitive function (memory) in mice were studied by trace fear conditioned reflex, and the effects of systemic inflammation and behavior after operation were measured by enzyme-linked immunosorbent assay (ELISA), the concentrations of CD163 and tumor necrosis factor-α (TNF-α) were measured. The expression of macrophage phenotype was observed by immunofluorescence staining, the expression levels of M1 and M2 markers mRNA were detected by real-time fluorescence quantitative PCR (RT-PCR), and the expression levels of FoxO1 and p21 were analyzed by immunoblotting (Western blot).
Compared with the control group, the freezing time in the anesthesia group was lower than that in the control group (P<0.01), indicating that sevoflurane anesthesia led to the decrease of cognitive ability. The blood concentrations of CD163 and TNF-α increased significantly at 24 h after the operation with sevoflurane anesthesia (P<0.05). Fluorescence microscopic observation showed that M2 was the main type of macrophages in normal tissues, while M1 and M2 phenotypes were highly expressed in sevoflurane anesthetized tissues at the same time, especially in M1 phenotypes (P<0.01). The polarization of macrophages in the anesthetic group showed the high level of M1 mRNA, and the expression levels of TNF-α, monocyte chemotactic protein 1(MCP-1) and Interleukin-6 (IL-6)mRNA in the anesthetic group were significantly higher than those in the control group (P<0.05). The expression levels of M2 mRNA such as transforming growth factor-β (TGF-β) and IL-10 were significantly lower than those in the control group (P<0.05). Compared with the control group, the expression of FoxO1 and p21 protein in the anesthesia group was significantly lower than that in the control group with a significant statistical difference (P<0.01).
This study offers a theoretical basis and insight for further understanding of the prevention and treatment of cognitive dysfunction induced by anesthetic drugs.
本研究旨在探讨叉头框 O1 和 p21 介导的巨噬细胞极化在七氟醚诱导术后认知功能障碍中的作用。
共纳入 30 只健康小鼠，随机分为两组：对照组（未行任何处理）和麻醉组（吸入七氟醚处理）。通过痕迹恐惧条件反射研究七氟醚对小鼠认知功能（记忆）的影响，通过酶联免疫吸附试验（ELISA）测定术后全身炎症和行为的影响，测定 CD163 和肿瘤坏死因子-α（TNF-α）的浓度。通过免疫荧光染色观察巨噬细胞表型的表达，实时荧光定量 PCR（RT-PCR）检测 M1 和 M2 标志物 mRNA 的表达水平，免疫印迹（Western blot）分析 FoxO1 和 p21 的表达水平。
与对照组相比，麻醉组的冻结时间明显低于对照组（P<0.01），表明七氟醚麻醉导致认知能力下降。七氟醚麻醉术后 24 h 时，血中 CD163 和 TNF-α 浓度明显升高（P<0.05）。荧光显微镜观察发现，正常组织中以 M2 型巨噬细胞为主，而七氟醚麻醉组织中同时高表达 M1 和 M2 表型，尤以 M1 表型为主（P<0.01）。麻醉组巨噬细胞极化表现为 M1 mRNA 水平升高，麻醉组 TNF-α、单核细胞趋化蛋白 1（MCP-1）和白细胞介素-6（IL-6）mRNA 表达水平明显高于对照组（P<0.05）。转化生长因子-β（TGF-β）和白细胞介素-10（IL-10）等 M2 mRNA 表达水平明显低于对照组（P<0.05）。与对照组相比，麻醉组 FoxO1 和 p21 蛋白表达明显低于对照组，差异有统计学意义（P<0.01）。
本研究为进一步了解麻醉药物引起的认知功能障碍的预防和治疗提供了理论依据和思路。
