敲低巨噬细胞移动抑制因子表达对人肺成纤维细胞增殖及迁移能力的影响  

Effect of MIF knocking-down on the proliferation and migration of human lung fibroblasts

作　　者：罗益锋[1] 易慧[1] 黄鑫炎[1] 谭卫平[1] 郭禹标[1] 谢灿茂[1] LUO Yifeng;YI Hui;HUANG Xinyan;TAN Weiping;GUO Yubiao;XIE Canmao(Division of Pulmonary and Critical Care Medicine,the First Affiliated Hospital of Sun Yat?sen University,Institute of Respiratory Diseases,Sun Yat?sen University,Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院呼吸与危重症医学科,中山大学呼吸病研究所,广州510080

出　　处：《实用医学杂志》2020年第6期716-721,共6页The Journal of Practical Medicine

基　　金：广东省科技计划项目(编号:2014A020212152)。

摘　　要：目的观察敲低巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)表达对人肺成纤维细胞(HFL1)增殖及迁移能力的影响,探讨MIF是否能作为抗纤维化治疗的新靶点。方法对照组采用细胞株HFL1-sh-NC。实验组采用MIF抑制的慢病毒感染HFL1细胞,构建MIF抑制细胞株HFL1-sh-MIF。荧光显微镜下观察细胞感染率,qRT-PCR法检测MIF mRNA表达量;CCK8检测敲低MIF表达后在体外对HFL1细胞增殖的影响,以光密度(OD)反映细胞增殖情况;Woundhealing法分析敲低MIF表达对HFL1细胞迁移能力的影响,以迁移率反映细胞的迁移能力。结果荧光结果显示MIF抑制慢病毒感染HFL1细胞后荧光率达95%以上,qRT-PCR验证结果显示,与对照组相比,实验组的MIF mRNA表达水平显著下降(P<0.001),说明MIF抑制慢病毒载体感染成功。CCK8的OD测量结果显示,4 h时两组差异无统计学意义(P>0.05);24、48和72 h时实验组的增殖显著下降(P<0.01)。Woundhealing结果显示,24 h时实验组和对照组HFL1细胞的迁移率差异无统计学意义(P>0.05);30 h时与对照组相比,实验组迁移率显著降低(P<0.01)。结论敲低MIF表达可显著抑制HFL1的增殖及迁移能力,从而减轻肺纤维化。MIF有望成为抗纤维化治疗的新靶点。Objective To investigate the impact of knocking down macrophage migration inhibitory factor(MIF)expression on proliferation and migration of human lung fibroblasts(HFL1)and explore the potentiality of MIF as a new therapeutic target for anti-fibrosis therapy.Methods Cell line HFL1-sh-NC was used to serve as a control group.Cell line in the experimental group was constructed by HFL1 cells with lentiviral infection,which was termed as HFL1-sh-MIF.The infected lentivirus were inhibited by MIF beforehand.The infection rate in HFL1 cells were assessed by fluorescence microscope.MIF mRNA expression in the cell line was quantified by qRT-PCR.CCK8 was evaluated to determine the influence on the proliferation of HFL1 cell line in vitro by knock-down MIF expression through the optical density(OD)measurement.The influence on the migration of HFL1 by knocking down MIF expression was assessed through the migration rate assessment by using Wouldhealing method.Results The infection rate by MIF-inhibited lentivirus in HFL1 cells was over 95%as determined by fluorescence microscope.Compared with the controlled group,MIF mRNA expression was significantly decreased in the experimental group(P<0.001)by q RTPCR.This demonstrated that the knock-down MIF HFL1 cell line by MIF-inhibited lentivirus infection was constructed successfully.The CCK8 OD measurement has shown that at 4 th hour,the proliferation rate was not affected by knockingdownMIFexpression(P>0.05);at 24 th hr,48 th hr and 72 nd hour,the proliferation rate was significantly impaired by knocking down MIF expression,as comparing the OD between control group and the experimental group.In terms of migration,at 24 th hour,there is no change on the migration rate between the experimental group and control group(P>0.05).At 30 th hour,there is a significant reduction of migration rate in the experimental group(P<0.01).Conclusions Knocking down MIF expression can significantly inhibit the proliferation and migration of HFL1.This in turn will improve the pulmonary fibrosis.MIF will

关 键 词：巨噬细胞移动抑制因子 人肺成纤维细胞 增殖 迁移 

分 类 号：R563.9[医药卫生—呼吸系统]