摘要
目的探讨细胞外信号调节蛋白激酶-5(ERK5)对人肺成纤维细胞系(HLFs)自噬、凋亡和增殖的调控及对成纤维细胞表型转化的影响。方法转化生长因子β1(TGF-β1)诱导HLFs建立肺纤维化细胞模型,过表达ERK5基因(ERK5)后通过Western blot检测自噬、凋亡、增殖相关因子和表型转化标志因子α-SMA的表达,通过细胞增殖/毒性检测(CCK8)试剂盒检测细胞增殖率。结果经TGF-β1处理后,与对照组相比,ERK5组细胞α-SMA、P62蛋白表达水平升高,LC3Ⅱ、Beclin-1蛋白表达水平降低;促凋亡蛋白Cleaved-Caspase-3表达水平降低,抗凋亡蛋白Bcl-2和增殖细胞核抗原PCNA表达水平升高,CCK8检测到细胞增殖率升高。结论在TGF-β1诱导的HLFs表型转化过程中,ERK5高表达促进细胞增殖、抑制细胞自噬和凋亡。
Objective To investigate the regulation of extracellular signal-regulated protein kinase-5(ERK5)on autophagy,apoptosis and proliferation of human lung fibroblasts(HLFs)and its influence on the phenotypic transformation of fibroblasts.Methods TGF-β1 induced HLFs to establish the pulmonary fibrotic cell model.After overexpressing ERK5 gene(ERK5),the expressions of autophagy,apoptosis,proliferation-related factors and phenotypic transformation markerα-SMA were detected by Western blot,and the cell proliferation rate was detected by the cell proliferation/toxicity detection kit(CCK8).Results After the treatment with TGF-β1,compared with the control group,the expression levels ofα-SMA and p62 proteins in the ERK5 group were increased,and the expression levels of LC3Ⅱand Beclin-1 proteins were decreased;the expression level of the pro-apoptotic protein cleaved-caspase-3 was decreased,and the expression levels of anti-apoptosis protein Bcl-2 and proliferating cellular nuclear antigen PCNA were increased,and the increase of cell proliferation rate was detected by CCK8.Conclusion During the phenotypic transformation process of HLFs induced by TGF-β1,the high expression of ERK5 promotes the cell proliferation and inhibits the cellular autophagy and apoptosis.
作者
华晓敏
王昌明
HUA Xiaomin;WANG Changming(Department of Respiratory Medicine,Affiliated Guilin Municipal People′s Hospital,Guilin Medical University,Guilin,Guangxi 541000,China)
出处
《重庆医学》
CAS
2021年第14期2362-2365,2370,共5页
Chongqing medicine
