摘要
目的探讨抑制HOXB7对TGF-β1诱导的肺泡上皮细胞纤维化的影响及可能的相关机制。方法 qRT-PCR检测肺纤维化组织中HOXB7的mRNA表达量;TGF-β1诱导A549纤维化;转染HOXB7siRNA到A549细胞,抑制HOXB7;倒置相差显微镜观察细胞形态变化;qRT-PCR检测A549细胞HOXB7、E-cadherin、α-SMA和COL1A1的mRNA表达量;Western blot检测A549细胞HOXB7、E-cadherin、α-SMA、bFGF和COL1A1蛋白表达量。细胞分组:control组(空白对照)、TGF-β1组(TGF-β1诱导)、β1-HOXB7 si组(TGF-β1诱导后转染HOXB7 siRNA)、β1-non-spe组(TGF-β1诱导后转染non-specific siRNA)、β1-HOXB7si-bFGF组(bFGF处理的β1-HOXB7 si组)。结果肺纤维组织HOXB7的mRNA表达量显著升高;细胞转染实验中HOXB7的mRNA和蛋白表达量均显著降低;降低HOXB7表达量显著抑制A549向细胞纤维化形态改变;抑制HOXB7显著升高E-cadherin mRNA和蛋白表达量,且降低α-SMA和COL1A1的mRNA和蛋白表达量;抑制HOXB7显著降低bFGF的蛋白表达量,且bFGF孵育细胞可有效反转抑制HOXB7对细胞E-cadherin、α-SMA和COL1A1表达量的影响。结论抑制HOXB7可通过调控bFGF有效降低TGF-β1诱导的肺泡细胞EMT过程,进而抑制其纤维化。
Objective To explore the effect of HOXB7 inhibition on TGF-β1-induced alveolar epithelial cell fibrosis and the related mechanism.Methods The mRNA expressions of HOXB7 in pulmonary fibrosis tissues were measured by qRT-PCR. TGF-β1 was used to induce the fibrosis of A549 cells. HOXB7 siRNA was transfected into A549 cells to down-regulate HOXB7 expression. Cell morphology changes were observed using inverted phase contrast microscope. The m RNA expressions of HOXB7,E-cadherin,α-SMA and COL1 A1 in A549 cells were measured by qRT-PCR. The protein expressions of HOXB7,E-cadherin,α-SMA,bFGF and COL1 A1 in A549 cells were detected by Western blot method. The cells were grouped as follows:control group(blank control),TGF-β1 group(TGF-β1 abduction),β1-HOXB7 si group(HOXB7 siRNA transfection after TGF-β1 abduction),β1-non-spe group(non-specific siRNA transfection after TGF-β1 abduction),β1-HOXB7 si-bFGF group(β1-HOXB7 si group with bFGF treatment).Results Both of the mRNA and protein expressions of HOXB7 decreased signifi-cantly in cells transfection. The decrease of the expression of HOXB7 significantly inhibited cell morphology change of A549 cells to fibrosis. The inhibition of HOXB7 significantly increased the mRNA and protein expression of E-cadherin,but decreased the m RNA and protein expression of α-SMA and COL1 A1. HOXB7 inhibition obviously reduced the protein expression of bFGF,and bFGF incubation effectively reversed the effects of HOXB7 inhibition on E-cadherin,α-SMA and COL1 A1 expression levels in A549.Conclusion HOXB7 inhibiti on reduces EMT pro-cess TGF-β1-induced through the regulation of bFGF,and further control the fibrosis in alveolar cell.
作者
周淼
李风雷
张海龙
孙俊波
ZHOU Miao;LI Fenglei;ZHANG Hailong;SUN Junbo(Department of Pulmonary Disease,Third Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China)
出处
《实用医学杂志》
CAS
北大核心
2018年第24期4037-4041,共5页
The Journal of Practical Medicine
基金
国家中医临床研究基地业务建设科研专项(编号:JDZX2015158)
河南中医学院博士科研基金(编号:BSJJ2015-27)
