摘要
目的 探讨抗TL1A单克隆抗体是否可通过影响结节病中巨噬细胞的极化水平,减轻结节病肉芽肿的形成。方法 采用随机分组法将32只C57BL/6小鼠分成空白对照组、结节病组、结节病+TL1A Ab治疗组和结节病+地塞米松治疗组(n=8)。通过肺组织病理学染色(HE染色)观察各组小鼠肺部肉芽肿的形成;通过流式细胞术检测各组小鼠脾脏单个核细胞(PBMC)中M1/M2型巨噬细胞的水平。提取各组小鼠股骨和胫骨内的原代骨髓源性巨噬细胞(BMDM),培养并分别诱导各组BMDM向M1或M2型巨噬细胞方向分化,使用流式细胞术与qPCR分别测定各组小鼠BMDM中M1型巨噬细胞和M2型巨噬细胞的细胞数目和相关因子的mRNA表达情况。结果 与结节病组相比,结节病+TL1A Ab组小鼠肺组织中肉芽肿的数目明显减少(P<0.001)。相较于空白对照组,结节病组PBMC中M1型巨噬细胞水平增高(P<0.001),给予TL1A Ab后可逆转这一现象(P<0.001);结节病+TL1A Ab组PBMC中M2型巨噬细胞的水平高于结节病组(P<0.001)。各组小鼠BMDM经诱导分化后,结节病+TL1A Ab组M1型巨噬细胞极化水平较结节病组下降(P<0.001);而TL1A Ab组M2型巨噬细胞极化水平高于结节病组(P<0.001)。结论 抗TL1A单克隆抗体通过调控结节病中巨噬细胞的极化水平而抑制结节病中的肉芽肿性炎。
Objective To investigate whether anti-TL1A monoclonal antibody(TL1A Ab)could mitigate granuloma formation in sarcoidosis by modulating the polarization of macrophages.Methods 32 C57BL/6 mice were randomly divided into four groups:the control group,the sarcoidosis group,the sarcoidosis+TL1A Ab treatment group,and the sarcoidosis+dexamethasone treatment group(n=8).Granuloma formation in mouse lungs was observed through histopathological staining(HE staining).Flow cytometry was used to assess the levels of M1/M2 macrophages in splenic mononuclear cells(PBMCs).Bone marrow-derived macrophages(BMDMs)were isolated from mouse femurs and tibias,cultured,and induced to differentiate into M1 or M2 macrophages.Flow cytometry and qPCR were used to determine the cell counts and mRNA expression of M1 and M2 macrophage-related factors in mouse BMDMs from each group.Results Compared with the sarcoidosis group,the number of granulomas in the lung tissue of mice in the TL1A Ab group was significantly reduced(P<0.001).Compared with the control group,the sarcoidosis group showed an elevated level of M1 macrophages in PBMCs(P<0.001),which was reversed upon administration of TL1A Ab(P<0.001).The TL1A Ab group exhibited higher levels of M2 macrophages in PBMCs compared to the sarcoidosis group(P<0.001).After inducing differentiation of mouse BMDMs,the polarization level of M1 macrophages in the TL1A Ab group was significantly decreased compared to the sarcoidosis group(P<0.001),and the levels of M1 macrophage-related cytokines,including iNOS,IL-10,and TNF-αwere all lower than those in the sarcoidosis group(all P<0.05).Conversely,the polarization level of M2 macrophages in the TL1A Ab group was higher than in the sarcoidosis group(P<0.001),and the levels of M2 macrophage-related cytokines,including MRC-1,IL-10,and TGF-βwere elevated compared with the sarcoidosis group(all P<0.05).The expression level of MMP-9 in M1 macrophages in the TL1A Ab group was significantly decreased compared with the sarcoidosis group(P<0.01).Conclusion Anti-T
作者
马成星
魏佳
单佳佳
郑颖
温艳婷
丁晶晶
MA Chengxing;WEI Jia;SHAN Jiajia;ZHENG Ying;WEN Yanting;DING Jingjing(Department of Respiratory and Critical Care Medicine,Nanjing Drum Tower Hospital,The Affiliated Hospital of Nanjing University Medical School,Nanjing,Jiangsu 210008,China;Department of Respiratory and Critical Care Medicine,Nanjing Drum Tower Hospital,Clinical College of Nanjing University of Chinese Medicine,Nanjing,Jiangsu 210008,China;Department of Basic Medicine,Center of Translational Medicine,Jiangsu Key Laboratory of Molecular Medicine,Nanjing University Medical School,Nanjing,Jiangsu 210008,China)
出处
《临床肺科杂志》
2024年第8期1146-1151,1156,共7页
Journal of Clinical Pulmonary Medicine
基金
国家自然科学基金面上项目(No.82170077)
南京市医药卫生科研课题(No.YKK20051)。
