摘要
目的 探讨白细胞介素6(IL-6)对MH-S小鼠肺泡巨噬细胞吞噬功能的影响及其相关机制。方法 脂多糖(LPS)经气道滴入激发构建小鼠急性肺损伤(ALI)模型,ELISA检测支气管肺泡灌洗液(BALF)中IL-6的含量。体外培养MH-S细胞,在信号转导子与转录激活子3 (STAT3)抑制剂Stattic(5μmol/L)存在与否的情况下,再加入IL-6(10 ng/mL~500 ng/mL)刺激6 h,加入荧光微球孵育2 h后,采用流式细胞术检测MH-S细胞吞噬荧光微球情况;Western blot法检测磷酸化的Janus激酶2(p-JAK2)、磷酸化的STAT3(p-STAT3)、肌动蛋白相关蛋白2(Arp2)、纤维型肌动蛋白(F-actin)的表达水平。结果 鼻腔滴入LPS后,小鼠BALF中IL-6含量显著升高。随着IL-6刺激剂量的增加,MH-S细胞吞噬荧光微球的作用增强,Arp2、 F-actin蛋白的表达水平升高。100 ng/mL IL-6刺激MH-S细胞后,p-JAK2和p-STAT3蛋白的表达水平升高。阻断MH-S细胞STAT3信号后,IL-6促进细胞吞噬的效应完全消失,IL-6诱导的Arp2、 F-actin蛋白表达增加被抑制。结论 IL-6通过激活JAK2/STAT3信号通路促进MH-S细胞Arp2、 F-actin蛋白的表达增强吞噬功能。
Objective To investigate the effect of interleukin-6(IL-6)on the phagocytosis of MH-S alveolar macrophages and its related mechanisms.Methods A mouse acute lung injury(ALI)model was constructed by instilling lipopolysaccharide(LPS)into the airway.ELISA was used to detect the content of IL-6 in bronchoalveolar lavage fluid(BALF).In vitro cultured MH-S cells,in the presence or absence of signal transducer and activator 3 of transcription(STAT3)inhibitor Stattic(5μmol/L),IL-6(10 ng/mL~500 ng/mL)was added to stimulate for 6 hours,and then incubated with fluorescent microspheres for 2 hours.The phagocytosis of MH-S cells was detected by flow cytometry.Western blot analysis was used to detect the expression levels of phosphorylated Janus kinase 2(p-JAK2),phosphorylated STAT3(p-STAT3),actin-related protein 2(Arp2)and filamentous actin(F-actin).Results The content of IL-6 in BALF was significantly increased after the mice were injected with LPS through the airway.With the increase of IL-6 stimulation concentration,the phagocytic function of MH-S cells was enhanced,and the expression levels of Arp2 and F-actin proteins in MH-S cells were increased.The expression levels of p-JAK2 and p-STAT3 proteins increased in MH-S cells stimulated with IL-6(100 ng/mL).After blocking STAT3 signaling,the effect of IL-6 in promoting phagocytosis of MH-S cells disappeared completely,and the increased expression of Arp2 and F-actin proteins in MH-S cells induced by IL-6 was also inhibited.Conclusion IL-6 promotes the expression of Arp2 and F-actin proteins by activating the JAK2 /STAT3 signaling pathway,thereby enhancing the phagocytic function of MH-S cells.
作者
华梦晴
高培宇
方芳
苏浩宇
宋传旺
HUA Mengqing;GAO Peiyu;FANG Fang;SU Haoyu;SONG Chuanwang(Department of Immunology,School of Laboratory Medicine,Bengbu Medical College,Bengbu 233030,China;Department of Immunology,Anhui Province Key Laboratory of Immunology in Chronic Diseases,Bengbu 233030,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2024年第1期13-18,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
安徽高校自然科学研究项目重大项目(KJ2020ZD49)
安徽高校自然科学研究项目重点项目(KJ2020A0583)
蚌埠医学院自然科学重点项目(2020byzd024)
蚌埠医学院省级重点科研平台开放课题基金项目(KLICD-2022-D2)
2021年安徽省大学生创新创业训练计划项目(S202110367103)。
