激活素A对L929成纤维细胞迁移、侵袭及细胞因子分泌的影响  

Effects of activin A on cytokine production and invasion of L929 fibroblasts

作　　者：张钰莹 单颖 ZHANG Yuying;SHAN Ying(Department of Immunology,Jinzhou Medical University,Jinzhou 121001,China)

机构地区：[1]锦州医科大学免疫学教研室,121001

出　　处：《免疫学杂志》2022年第1期22-29,共8页Immunological Journal

摘　　要：目的探讨激活素A(activin A)对小鼠成纤维细胞系L929细胞迁移、侵袭及细胞因子分泌的影响。方法采用5-溴-2’-脱氧尿嘧啶核苷(5-bromo-2’-deoxyuridine,BrdU)掺入实验检测L929细胞增殖,酶联免疫吸附实验(ELISA)检测L929细胞培养上清中细胞因子水平,基质胶transwell侵袭实验分析L929细胞迁移、侵袭,实时细胞分析技术(RTCA)检测L929细胞黏附,Western blot分析细胞内信号蛋白表达水平。结果与对照组相比,激活素A处理对L929细胞增殖没有影响,对L929细胞分泌TNF-α、IL-1β及IL-6等促炎细胞因子也没有影响,但却显著促进纤维化诱导因子TGF-β1分泌。激活素A还能显著增加L929细胞穿透基质胶迁移、侵袭细胞数量(P<0.01),并促进L929细胞黏附,添加阻断剂FST可以显著减弱激活素A诱导的L929细胞侵袭和促进黏附作用。激活素A处理L929细胞可以显著增加p-ERK和p-JNK蛋白水平,但对p-Smad3蛋白水平没有明显影响。结论激活素A可以促进L929细胞分泌TGF-β1、诱导L929细胞侵袭和促进细胞黏附,其作用可能与非Smad3依赖的ERK和JNK信号有关。This study was designed to investigate the effects of activin A on invasion and cytokine secretion of mouse fibroblast cell line L929 cells.After treatment with activin A,the proliferation of L929 cells was determined by BrdU method,the cytokine levels in the supernatant of cultured L929 cells were detected by ELISA,the invasion of L929 cells was examined by Matrigel-coated transwell invasion assay,the adhesion of L929 cells was analyzed by real-time cell-based assay(RTCA),and the levels of intracellular signal proteins were examined by Western blotting.Data showed that compared with the control group,activin A treatment had no effect on the proliferation of L929 cells and also did not alter the secretion of pro-inflammatory cytokines TNF-α,IL-1β1 and IL-6,but significantly promoted the fibrosis-inducing factor TGF-β1 production.Activin A also significantly increased the number of L929 cells invading through Matrigel-coated transwell(P<0.01)and promoted L929 cell adhesion,while the addition of FST as antagonist of activin A could significantly reduce L929 cell invasion and adhesion induced by activin A.Furthermore,activin A treatment significantly increased the levels of p-ERK and p-JNK proteins in L929 cells,but had no significant effect on the level of p-smad3 protein.In conclusion,activin A can promote TGF-β1 secretion by L929 cells,induce L929 cell invasion and increase L929 cell adhesion,and the mechanism may related to Smad3-independent ERK and JNK signaling.

关 键 词：激活素A L929细胞 细胞因子 侵袭 

分 类 号：R392[医药卫生—免疫学]