红色诺卡菌细胞壁骨架对人中性粒细胞生物学功能的调节作用  

Regulatory effects of the Nocardia rubra cell wall skeleton on the biological function of human neutrophils

作　　者：杨云稀 黄佳敏 刘璐 李林斌 郑淳方 周于莹 孙炳伟 Yang Yunxi;Huang Jiamin;Liu Lu;Li Linbin;Zheng Chunfang;Zhou Yuying;Sun Bingwei(Department of Burns and Plastic Surgery,Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou 215002,China)

机构地区：[1]南京医科大学附属苏州医院烧伤整形科,苏州215002

出　　处：《中华烧伤与创面修复杂志》2023年第7期625-632,共8页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金联合基金重点项目(U21A20370);国家自然科学基金面上项目(82072217,81772135);江苏省自然科学基金项目(BK20201178)。

摘　　要：目的探讨红色诺卡菌细胞壁骨架(Nr-CWS)对人中性粒细胞生物学功能的调节作用及其机制。方法采用实验研究方法。2022年5―10月招募于苏州市体检中心体检的成年健康志愿者15名(男7名、女8名,年龄24~45岁),采集外周静脉血,采用免疫磁珠分选法提取中性粒细胞。将细胞分为不进行任何处理的正常对照组、仅用终质量浓度为60 ng/mL Nr-CWS处理的单纯Nr-CWS组、仅用终质量浓度1μg/mL内毒素/脂多糖(LPS)刺激的单纯LPS组、用同前LPS刺激后再用Nr-CWS处理的LPS+Nr-CWS组。培养1 h,采用改良后的琼脂糖趋化模型检测中性粒细胞的趋化距离、趋化细胞百分比、趋化指数、最大趋化速度、趋化功能评分;采用流式细胞术检测发生吞噬的细胞占比与荧光强度,活性氧水平,颗粒蛋白CD35、CD66b、CD63的蛋白表达水平,以及细胞培养上清液中炎症因子白细胞介素2(IL-2)、IL-4、IL-6、IL-10、IL-17A、肿瘤坏死因子α(TNF-α)、γ干扰素浓度。以上实验各组样本数均为15。对数据行析因设计方差分析与独立样本t检验。结果培养1 h,正常对照组、单纯Nr-CWS组、单纯LPS组、LPS+Nr-CWS组细胞趋化功能评分分别为15.0、(14.5±0.5)、(1.5±0.5)、(12.0±1.5)分;与正常对照组比较,单纯LPS组与LPS+Nr-CWS组细胞趋化距离、趋化细胞百分比、趋化指数、最大趋化速度与趋化功能评分均显著降低(t值分别为18.36、18.88、54.28、18.36、46.77,10.58、14.74、6.84、10.58、4.24,P<0.05);与单纯LPS组比较,LPS+Nr-CWS组细胞上述5项趋化功能指标均显著升高(t值分别为11.47、14.65、11.62、11.47、13.75,P<0.05)。培养1 h,与正常对照组比较,单纯Nr-CWS组发生吞噬的细胞占比与荧光强度均显著升高(t值分别为6.86、6.73,P<0.05),单纯LPS组(t值分别为7.35、22.72,P<0.05)与LPS+Nr-CWS组(t值分别为21.37、13.10,P<0.05)发生吞噬的细胞占比与荧光强度均明显降低。培养1 h,与正常对照组�Objective To investigate the regulatory effects and mechanism of Nocardia rubra cell wall skeleton(Nr-CWS)on the biological function of human neutrophils.Methods The experimental research method was used.Fifteen healthy adult volunteers(7 males and 8 females,aged 24 to 45 years)were recruited from Suzhou Physical Examination Center for physical examination from May to October 2022,the peripheral venous blood was collected,and neutrophils were extracted by immunomagnetic bead sorting.The cells were divided into normal control group without any treatment,Nr-CWS alone group treated with Nr-CWS of final mass concentration 60 ng/mL alone,endotoxin/lipopolysaccharide(LPS)alone group stimulated with LPS of final mass concentration 1μg/mL alone,and LPS+Nr-CWS group stimulated with LPS first and then treated with Nr-CWS as before.After 1 h of culture,the chemotaxis distance,chemotactic cell percentage,chemotactic index,maximum chemotactic speed,and chemotactic function score of neutrophils were detected using the modified agarose chemotactic model;the proportion and fluorescence intensity of phagocytosis cells,the level of reactive oxygen species(ROS),the protein expression levels of granular protein CD35,CD66b,and CD63,and the concentrations of inflammatory cytokines of interleukin 2(IL-2),IL-4,IL-6,IL-10,IL-17A,tumor necrosis factor alpha(TNF-α),and interferon-γin cell culture supernatant were detected by flow cytometry.The number of samples in each group in the above experiments was 15.Data were statistically analyzed with analysis of variance for factorial design and independent sample t test.Results After 1 h of culture,the chemotactic function score of cells in normal control group,Nr-CWS alone group,LPS alone group,and LPS+Nr-CWS group were 15.0,14.5±0.5,1.5±0.5,12.0±1.5,respectively.Compared with those in normal control group,the chemotaxis distance,chemotactic cell percentage,chemotactic index,maximum chemotactic speed,and chemotactic function score of cells were significantly decreased in LPS alone group a

关 键 词：感染 吞噬作用 趋化作用 中性粒细胞 生物学功能 红色诺卡菌细胞壁骨架 

分 类 号：R392[医药卫生—免疫学]