CD40-CD40L交联系统在矽肺纤维化形成中的作用机制  

The Function of CD40/CD40L Pathway in Silicosis Fibrosis

作　　者：王淑娟[1] 胡魁[1] 韩静茵[1] 干晓瑜[1] 娄懿[1] 李国辉[1] Wang Shujuan;Hu Kui;Han Jingyin;Gan Xiaoyu;Lou Yi;Li Guohui(Department of Occupational Disease,Zhejiang Integrated Traditional and Western medicine Hospital,Hangzhou 310003,China)

机构地区：[1]浙江省中西医结合医院职业病科,浙江杭州310003

出　　处：《中华劳动卫生职业病杂志》2020年第2期96-100,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：杭州市卫计委科技计划项目(2016A36);杭州市科技发展计划项目(20170533B61)。

摘　　要：目的探讨CD40-CD40配体(CD40L)交联系统在矽肺纤维化形成中的作用.方法选取2016至2018年在浙江省中西医结合医院职业病科门诊及住院收治的患者共64例,根据病例诊断将其分为矽肺组、对照组,每组各32人.收集两组患者肺泡灌洗液(BALF),用流式细胞术检测T淋巴细胞表面CD40L蛋白表达的水平;用ELISA法测定BALF中白细胞介素(IL-8、IL-6)、干扰素(INF-γ)和单核细胞趋化蛋白(MCP-1)的含量.将两组BALF与人胚肺成纤维细胞系HFL-1细胞共培养,用免疫印迹试验(Western Blot)法检测Ⅰ型胶原蛋白(CollagenⅠ)的表达水平;用免疫荧光法检测α-平滑肌动蛋白(α-SMA)的表达情况.结果与对照组BALF分离得到的T淋巴细胞比较,矽肺组BALF分离得到的T淋巴细胞表面CD40L的表达明显升高,差异有统计学意义(P<0.05).与对照组比较,矽肺组BALF中IL-8、IL-6、INF-γ、MCP-1的含量均明显升高,差异有统计学意义(P<0.05).与对照组BALF加入培养基培养HFL-1细胞比较,矽肺组肺泡灌洗液加入培养基培养HFL-1细胞的CollagenⅠ蛋白表达量、α-SMA荧光表达均明显升高,差异均有统计学意义(P<0.05).结论CD40-CD40L交联系统可促进T细胞活化,释放炎症因子,促进成纤维细胞合成CollagenⅠ和α-SMA,使肺纤维组织增生,导致矽肺纤维化的形成.Objective To investigate the role of CD40/CD40L Pathway in the formation of silicosis fibrosis.Methods Totally 64 inpatients were recruited and assigned to the silicosis group and the control group,23 in each group.The alveolar lavage fluid was collected from all patients and isolated.The expression of CD40L protein was detected by Flow Cytometry.The level of IL-8、The IL-6、INF-γand MCP-1 was detected by ELISA.Two groups of BALF were co-cultured with HFL-1 cells,the expression of Collagen I andα-SMA was detected by Immunohistochemistry.Results Compared with the control group,CD40L was highly expressed on T lymphocyte cells in silicosis group(P<0.05),and the contents of IL-8、The IL-6、INF-γand MCP-1 in Silicosis group were significantly higher than those in control group(P<0.05).After co-culture of BALF and HFL-1 cells,the expression levels of Collagen I andα-SMA in Silicosis group were significantly higher than those in control group(P<0.05).Conclusion CD40-CD40L cross-linking system can promote the activation of T cells,release inflammatory factors,promote the synthesis of collagen I andα-SMA by fibroblasts,make the lung fibrous tissue proliferate,and lead to the formation of silicosis fibrosis.

关 键 词：矽肺 纤维化 CD40-CD40配体 Ⅰ型胶原蛋白 Α-平滑肌动蛋白 

分 类 号：R1[医药卫生—公共卫生与预防医学]