白葡奈氏菌片对慢阻肺小鼠辅助性T淋巴细胞17/调节性T淋巴细胞平衡的影响及其作用机制研究  

Effect of Staphylococcus and Neisseria tablets on Th17/Treg balance in mice with chronic obstructive pulmonary disease and its mechanism of action

作　　者：杨洋[1] 申思 罗湘 王志霞 张志强[1] YANG Yang;SHEN Si;LUO Xiang;WANG Zhi-xia;ZHANG Zhi-qiang(WardⅡ,Department of Respiratory and Critical Care Medicine,the First Affiliated Hospital of Xinxiang Medical College,Weihui 453100,Henan,CHINA)

机构地区：[1]新乡医学院第一附属医院呼吸与危重症医学科二病区,河南卫辉453100

出　　处：《海南医学》2025年第6期767-772,共6页Hainan Medical Journal

基　　金：河南省医学科技攻关计划联合项目(编号:LHGJ20210504)。

摘　　要：目的探讨白葡奈氏菌片对慢性阻塞性肺疾病(COPD)小鼠辅助性T淋巴细胞17(Th17)/调节性T淋巴细胞(Treg)平衡的影响及其可能的作用机制。方法将40只C57BL/6小鼠随机分为对照组(CON组)、慢阻肺模型组(COPD组)、白葡奈氏菌低剂量干预COPD组(SANTL组)、白葡奈氏菌高剂量干预COPD组(SANTH组),每组10只。使用8周烟熏方法构建COPD模型。SANTL组及SANTH组在造模成功后分别给予低剂量(10μg/kg,3次/d)及高剂量(20μg/kg,3次/d)白葡奈氏菌片混悬液灌胃4周,而COPD组给予对应体积生理盐水灌胃4周,结束后处死小鼠。通过苏木精-伊红(HE)染色评估肺组织病理学改变,酶联免疫吸附试验(ELISA)检测支气管肺泡灌洗液(BALF)中IL-17、IL-22、IL-10、IL-35、IL-6及肿瘤坏死因子(TNF-α)水平,流式细胞术分析脾脏Th17和Treg细胞占CD4+淋巴细胞比率(Th17%、Treg%),采用Spearman相关性分析法分析BALF中IL-6及TNF-α与IL-17、IL-22、IL-10、IL-35表达水平的相关性。结果与CON组比较,COPD组小鼠体质量增长明显降低,差异有统计学意义(P<0.05);HE染色肺组织呈现广泛炎症浸润、肺泡壁断裂等典型病理特征;与CON组比较,COPD组小鼠的Th17%、IL-17、IL-22、IL-6、TNF-α表达水平明显上升,而Treg%、IL-10及IL-35表达水平明显下降,差异均有统计学意义(P<0.05);经白葡奈氏菌片干预的两组小鼠Th17%[SANTL:(1.26±0.18)%;SANTH:(0.68±0.16)%]、IL-17[SANTL:(65.79±12.03)ng/L;SANTH:(46.52±4.87)ng/L]、IL-6[SANTL:(55.17±3.35)ng/L;SANTH:(48.16±3.51)ng/L]、TNF-α[SANTL:(217.71±19.26)ng/L;SANTH:(193.15±9.20)ng/L]表达水平均低于COPD组,但均高于CON组,其中SANTH组低于SANTL组,差异均有统计学意义(P<0.05);经白葡奈氏菌片干预的两组小鼠Treg%[SANTL:(7.98±1.11)%;SANTH:(9.25±0.73)%]、IL-10[SANTL:(9.31±0.67)ng/L;SANTH:(11.52±1.26)ng/L]及IL-35[SANTL:(12.99±1.69)ng/L;SANTH:(17.73±3.17)ng/L]表达水平均高于COPD组,但均低于CON组,其中SANTH组高于SANTL组Objective To investigate the effects of Staphylococcus and Neisseria tablets on the helper T lymphocyte 17(Th17)/regulatory T lymphocyte(Treg)balance in mice with chronic obstructive pulmonary disease(COPD)and the possible mechanisms involved.Methods Forty C57BL/6 mice were randomly divided into control group(CON group),COPD model group(COPD group),low-dose Staphylococcus and Neisseria tablets intervention group(SANTL group,10μg/kg,3 times/day),and high-dose Staphylococcus and Neisseria tablets intervention group(SANTH group,20μg/kg,3 times/day),with 10 mice in each group.The COPD model was established using an 8-week cigarette smoke exposure method.After successful modeling,SANTL and SANTH groups received Staphylococcus and Neisseria tablets suspension by gavage for 4 weeks,while the COPD group received saline.Lung pathology was assessed by hematoxylin-eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was used to determine the expression levels of the airway inflammation-related cytokines interleukin-17(IL-17),IL-22,IL-10,IL-35,IL-6 and tumour necrosis factor-α(TNF-α)in bronchoalveolar lavage fluid(BALF),flow cytometry was used to determine the ratio of splenic Th17 and Treg cells to CD4+lymphocytes(Th17%,Treg%),and spearman correlation analysis was used to analyze the correlation between IL-6,TNF-αand the expression levels of IL-17,IL-22,IL-10,IL-35 in BALF.Results Compared with the CON group,the COPD group showed significantly reduced body mass(P<0.05);HE staining revealed extensive inflammatory infiltration and alveolar wall rupture in the lung tissue of the COPD group;Compared with the CON group,the COPD group exhibited significantly increased Th17%,IL-17,IL-22,IL-6,and TNF-αlevels,while Treg%,IL-10,and IL-35 levels were significantly decreased(P<0.05).Staphylococcus and Neisseria tablets intervention(SANTL and SANTH groups)dose-dependently reduced Th17%,IL-17,IL-6,and TNF-α,while increasing Treg%,IL-10,and IL-35(all P<0.05):Th17%,SANTL(1.26±0.18)%,SANTH(0.68±0.16)%;IL-17,SANTL(65.79±12.03)

关 键 词：慢性阻塞性肺疾病 辅助性T淋巴细胞17/调节性T淋巴细胞平衡 白葡奈氏菌片 免疫调节 剂量依赖性 作用机制 

分 类 号：R-332[医药卫生]