摘要
目的 探讨激活N型乙酰胆碱受体对急性呼吸窘迫综合征(ARDS)小鼠白细胞介素-18 (IL-18)、PD-1和T淋巴细胞等的影响。方法 将6周龄健康清洁级雄性BALB/c小鼠60只按照完全数字随机法分为正常组(N组)、生理盐水对照组(NS组)、生理盐水+切断颈部两侧迷走神经组(NS+D组)、ARDS+切断颈部两侧迷走神经组(A+D组)、ARDS组(A组)、ARDS+切断迷走神经给予乙酰胆碱受体激动剂组(A+J组),每组10只。用荧光显微镜观察小鼠右肺下叶组织结构变化。用免疫印迹法检测各组小鼠左肺磷酸化核因子κB (p-NF-κB) P65和PD-1的水平,用酶联免疫吸附试验(ELISA)双抗体夹心法测定各组小鼠血清IL-18和PD-1含量,用流式细胞仪检测小鼠右肺中叶CD3^(+)、CD25^(+)Foxp3^(+)T淋巴细胞百分比。结果 荧光显微镜下,N组和NS组小鼠右肺下叶间质未发现炎症细胞浸润,NS+D组肺间质有少量炎症细胞浸润,而A组和A+D组小鼠肺间质有大量炎症细胞浸润、肺泡壁增厚、肺泡结构破坏,肺泡腔消失;A+J组小鼠右肺下叶间质炎症细胞浸润较少,肺泡腔有少许破损。A组和A+D组小鼠血清IL-18和PD-1水平高于其他4组,差异有统计学意义(P <0.05)。A组和A+D组小鼠左肺p-NF-κBP65和PD-1蛋白含量显著高于N组、NS组、J组(P <0.05);A组和A+D组小鼠肺泡灌洗液CD3^(+)、CD25^(+)Foxp3^(+)T淋巴细胞百分比高于其他3组,A+J组显著低于A+D组、A组(P <0.05)。结论 激活N型乙酰胆碱受体可通过T淋巴细胞负向调节作用直接抑制IL-18和p-NF-κBP65的释放,还可抑制肺组织PD-1的生成,从而抑制ARDS的炎症反应,减轻ARDS病理变化。
Objective To investigate the effect of activating the N-type acetylcholine receptor(nAChR) on interleukin-18(IL-18) and PD-1 in mice with acute respiratory distress syndrome(ARDS).Methods Sixty healthy male BALB/c mice(6 weeks of age) were divided into six groups:normal(N),normal saline control(NS),normal saline + bilateral vagectomy(NS+D),ARDS + segmentation of the vagus nerve on both sides of the neck(A+D),ARDS(A),ARDS + vagal amputation,and administration of an acetylcholine receptor agonist(A+J) groups.Each group included ten mice that were fed and housed under normal conditions.Structural changes in the right lower lung were observed using fluorescence microscopy;phosphorylated nuclear factor-κB protein 65(p-NF-κBP65) levels were assessed using Western blotting;serum IL-18 and PD-1 levels were assessed using enzyme-linked immunosorbent assay(ELISA) and the double antibody sandwich method;and the percentages of CD3~+ and CD25~+Foxp3~+T lymphocytes in the middle lobe of right lung were determined using flow cytometry.Results No inflammatory cell infiltration was observed in groups N and NS.The interstitial lobes in groups A and A+D showed severe inflammatory infiltration,thickening of the alveolar wall,destruction of the alveolar structure,and loss of the alveolar cavity.Serum IL-18 and PD-1 levels in groups A and A+D were significantly higher than those in the other four groups(P < 0.05).p-NF-κBP65 and PD-1 levels in groups A and A+D were significantly higher than those in groups N,NS,and A+J(P < 0.05).CD3~+ and CD25~+Foxp3~+T cells in groups A and A+D were significantly higher than those in the other four groups(P < 0.05).Conclusion Active nAChR can inhibit IL-18 and p-NF-κBP65 through the negative regulation of T lymphocytes,decrease PD-1 expression in lung tissues,and alleviate the pathological changes of ARDS.
作者
殷宗宝
余燕梅
刘凡
YIN Zongbao;YU Yanmei;LIU Fan(Emergency Department,Haikou Affiliated Hospital of Central South University Xiangya School of Medicine,Haikou 570208,China)
出处
《中国医科大学学报》
北大核心
2025年第2期133-138,共6页
Journal of China Medical University
基金
海南省自然科学基金(821RC759)。
关键词
急性呼吸窘迫综合征
N型乙酰胆碱受体
炎症
acute respiratory distress syndrome
nicotinic acetylcholine receptor
inflammation
