FoxO1和p21介导巨噬细胞极化对七氟烷减轻术后认知功能障碍的促进作用

Promoting Effect of Macrophage Polarization Mediated by FoxO1 and p21 of Sevoflurane on Alleviating Postoperative Cognitive Impairment

目的探讨FoxO1、p21介导巨噬细胞极化在七氟烷致术后认知功能障碍中的作用。方法 120只大鼠按随机数字表法分为对照组、手术组和麻醉组。对照组仅注射戊巴比妥钠麻醉,手术组和麻醉组行剖腹手术,麻醉组给予七氟烷。通过Morris水迷宫实验观察麻醉组大鼠在七氟烷致术后认知功能变化;以流式细胞术检测海马神经元凋亡率、胞质钙离子(Ca2+)浓度([Ca2+]i);利用qRT-PCR实验测定大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、 IL-6因子表达水平;Western blotting测定巨噬细胞FoxO1、p21蛋白表达;采用酶联免疫吸附测定(ELISA)法对M1、M2型巨噬细胞相关炎性因子表达进行测定。结果手术组术后各时间点逃逸潜伏期延长,平台象限停留时间缩短,穿越原平台的次数减少;麻醉组术后第1,3,5,7天潜伏期延长,平台象限停留时间缩短,穿越原平台的次数减少。与对照组比较,手术组海马神经元凋亡率显著增高(P<0.05);术后第1天海马神经元胞质[Ca2+]i浓度增加,术后第7天海马神经元胞质[Ca2+]i浓度降低;与对照组比较,手术组海马神经元胞质[Ca2+]i浓度显著增高(P<0.05),麻醉组TNF-α、IL-1β、IL-6表达水平均高于对照组,但差异无统计学意义(P>0.05),麻醉组FoxO1、p21蛋白表达量下降,但差异无统计学意义(P>0.05);与对照组比较,手术组M1型巨噬细胞标志物iNOS mRNA表达量显著降低(P<0.05),M2型巨噬细胞标志物IL-10、Arg-1 mRNA表达量显著增高(P<0.05),麻醉组M1型巨噬细胞标志物iNOS mRNA表达量显著增加(P<0.05),M2型巨噬细胞标志物IL-10、Arg-1 mRNA表达量显著降低(P<0.05)。结论 FoxO1、p21介导巨噬细胞极化对七氟烷致术后减轻认知功能障碍具有促进作用。

Objective To explore the role of macrophage polarization mediated by FoxO1 and p21 in sevoflurane-induced cognitive impairment after surgery.Methods Total 120 rats were randomly divided into control group,operation group and anesthesia group.The control group was anesthetized only with pentobarbital sodium,and the operation group and anesthesia group were operated by laparotomy,and the anesthesia group was given sevoflurane.The cognitive function of rats after sevoflurane-induced surgery was determined by Morris test.The apoptotic rate of hippocampal neurons and the concentration of cytoplasmic Ca 2+were detected by flow cytometry,and the expression levels of TNF-α,IL-1βand IL-6 in serum of rats were determined by qRT-PCR.The expression of FoxO1 and p21 protein in macrophages was measured by Western blotting,and the expression of macrophage-related inflammatory factors M1 and M2 were determined by ELISA.Results The escape latency of operation group was prolonged at all time points after operation,and the platform quadrant residence time was shortened,and the number of times of crossing the original platform was reduced in the operation group.In anesthesia group,the latency period was prolonged on the first,third,fifth,and seventh days after operation,and the platform quadrant residence time was shortened and the number of times of crossing the original platform was reduced.Compared with the control group,the apoptotic rate of hippocampal neurons in the operation group changed significantly(P<0.05).The[Ca 2+]concentration of hippocampal neuron cytoplasm increased on the first day after the operation,and decreased on the seventh day after the operation.Compared with the control group,the[Ca 2+]concentration of hippocampal neuron cytoplasm changed significantly in the operation group(P<0.05).In the anesthesia group(P<0.05).The expression levels of TNF-α,IL-1β,and IL-6 were higher than those in the control group,but there were no significant difference(P>0.05).The expression levels of FoxO1 and p21 in anesthesia group were lower than those in control group(P>0.05).Compared with control group,the expression levels of macrophage markers iNOS in operation group were significantly lower(P<0.05),and the expression levels of macrophage markers IL-10 and Arg-1 in M2 group were significantly lower(P<0.05).The expression of iNOS and IL-10 and Arg-1 were significantly increased in anesthesia group(P<0.05),but decreased in M2 group(P<0.05).Conclusion FoxO1 and p21 mediated macrophage polarization can promote the alleviation of sevoflurane-induced cognitive impairment.