二甲双胍通过FGFR4/MST1信号通路抑制皮肤瘢痕疙瘩成纤维细胞的上皮间质转化  

Metformin inhibits epithelial mesenchymal transformation of cutaneous keloid fibroblasts through FGFR4/MST1 signaling pathway

作　　者：张永红 李存涛 高明敏 张玉红 ZHANG Yonghong;LI Cuntao;GAO Mingmin;ZHANG Yuhong(Department of Dermatology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学附属郑州中心医院皮肤科,河南郑州450052

出　　处：《东南大学学报（医学版）》2022年第6期793-799,共7页Journal of Southeast University(Medical Science Edition)

基　　金：河南省医学科技攻关计划(联合共建)项目(LHGJ20191506)。

摘　　要：目的:探讨二甲双胍(MET)抑制低氧诱导的瘢痕疙瘩成纤维细胞(KFs)上皮间质转化(EMT)的作用机制。方法:KFs被分为低氧组和对照组,低氧组细胞在低氧环境下培养KFs以诱导EMT,对照组细胞常规培养。用MET或其溶剂生理盐水(生理盐水组)低氧诱导KFs;在MET和低氧条件下用纤维母细胞生长因子受体4(FGFR4)过表达载体(FGFR4-OE)或空载体(EV)处理KFs,或者在MET和低氧条件下用重组人巨噬细胞刺激1蛋白(rhMST1)处理KFs;低氧条件下的KFs分为生理盐水组、MET组、EV+MET组、FGFR4-OE+MET组、rhMST1+MET组。采用细胞计数试剂盒8(CCK-8)评估细胞增殖活力,迁移小室实验测量细胞迁移,蛋白免疫印迹法评估FGFR4、E-钙黏蛋白、波形蛋白和MST1的表达水平。结果:低氧组KFs的细胞增殖活力、迁移率及EMT较对照组明显增加。MET组的FGFR4、MST1表达以及低氧诱导的细胞增殖活力、迁移率及EMT标志物的表达量都较生理盐水组明显降低(P<0.05)。与MET组及EV+MET组比较,FGFR4-OE+MET组中FGFR4、MST1表达以及低氧诱导的细胞增殖活力、迁移率和EMT标志物均明显增高(P<0.05)。与MET组相比,rhMST1+MET组MST1表达及低氧诱导的细胞增殖活力、迁移率、EMT标志物的mRNA表达量均明显增高(P<0.05)。结论:MET通过抑制FGFR4/MST1信号通路来消除KFs中低氧诱导的EMT。Objective: To investigate the mechanism of metformin(MET) on hypoxia-induced keloid fibroblasts(KFs) epithelial-mesenchymal transition(EMT). Methods: KFs was divided into a hypoxic group and a control group. KFs in the hypoxic group were cultured in a hypoxic environment to induce EMT, while KFs in the control group were normal cultured. Additionally, KFs were treated with fibroblast growth factor receptor 4(FGFR4) overexpression vector(FGFR4-OE) or empty vector(EV) in the presence of MET and under hypoxic conditions, respectively. KFs were treated with recombinant human macrophage stimulation 1(rhMST1) protein in the presence of MET and hypoxia. KFs under hypoxia were divided into saline group, MET group, EV+MET group, FGFR4-OE+MET group and rhMST1+MET group. Cell proliferation viability was assessed by cell counting kit 8(CCK-8), and cell migration was measured by migration chamber assay. Expression levels of FGFR4, E-cadherin, vimentin and MST1 were assessed by western blotting. Results: Compared with the control group, the cell proliferation activity of KFs, migration rate and EMT in the hypoxia group were significantly increased. The expressions of FGFR4 and MST1, as well as hypoxia-induced cell proliferation activity, migration rate and EMT markers in the MET group were significantly lower than those in the normal saline group(P<0.05). Compared with the MET group and the EV+MET group, the expressions of FGFR4 and MST1, as well as the hypoxia-induced cell proliferation activity, migration rate and EMT markers in the FGFR4-OE+MET group were significantly increased(P<0.05). Compared with the MET group, the MST1 expression and hypoxia-induced cell proliferation activity, migration rate and EMT markers in the rhMST1+MET group were significantly increased(P<0.05). Conclusion: MET abrogates hypoxia-induced EMT in KFs by inhibiting the FGFR4/MST1 signaling pathway.

关 键 词：二甲双胍 上皮间质转化 纤维母细胞生长因子受体4 巨噬细胞刺激1 瘢痕疙瘩 

分 类 号：R758[医药卫生—皮肤病学与性病学]