TGF-β1和TNF-α协同刺激诱导人支气管上皮细胞优化上皮间质转化模型  

Synergy of TGF-β1 and TNF-α inducing human bronchial epithelial cells to optimize epithelia-mesenchymal transformation model

作　　者：何铭[1] 朱恺[1] 段洪涛[1] 李鹏程[2] 闫小龙[1] 张志培[1] 李小飞[1] 

机构地区：[1]第四军医大学唐都医院胸腔外科,西安710038 [2]第四军医大学唐都医院呼吸与危重症医学科,西安710038

出　　处：《中国胸心血管外科临床杂志》2017年第3期228-232,共5页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：国家自然科学基金资助(81370115)

摘　　要：目的探索转化细胞生长因子-β1(transforming growth factor-β1,TGF-β1)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)协同刺激诱导人气管上皮细胞(human bronchial epithelial cell,HBE)细胞,优化上皮间质转化细胞模型的效果。方法采用空白对照、TGF-β1 10 ng/ml、TNF-α10 ng/ml、TGF-β1 10 ng/ml+TNF-α10 ng/ml刺激诱导HBE细胞24 h后,观察细胞的变化。采用CCK8、细胞迁徙、蛋白印迹实验来实现。结果 TGF-β1+TNF-α协同诱导下大多数细胞从鹅卵石样形态变化为梭形、边界消失、间隙明显增宽。空白对照组、TGF-β1 10 ng/ml组、TNF-α10 ng/ml组、TGF-β1 10 ng/ml+TNF-α10 ng/ml组的细胞迁移距离分别为(420.06±10.38)μm、(499.86±34.00)μm、(514.93±10.56)μm和(569.68±33.58)μm。TGF-β1+TNF-α组与对照组、TGF-β1、TNF-α的差异有统计学意义(P<0.05)。TGF-β1+TNF-α协同诱导时,上皮标志物E-cadherin明显降低,间质标志物Vimentin明显升高。结论在HBE细胞中采用TGF-β1和TNF-α协同诱导可以优化细胞上皮间质转化模型。Objective To explore the effectiveness of the transforming growth factor-β1（TGF-β1） and tumor necrosis factor-α（TNF-α） inducing human bronchial epithelial（HBE） cells to optimize epithelia-mesenchymal transformation（EMT） model. Methods Blank control, TGF-β1 10 ng/ml, TNF-α 10 ng/ml, TGF-β1 10 ng/ml＋TNF-α 10 ng/ml induced human epithelial cells for 24 hours. Then the change of morphological alteration were observed by applying CCK8, cells migration assay and Western blot technique. Results When TGF-β1 plus TNF-α induced human epithelial cells for 24 hours, most of HBE cells traits changed including morphological alteration from cobblestone to fusiform, connection between cells vanishing, intercellular space broadening. In the experiments of checking cell migration capacity by the vitro scratch test, the group spacing was 420.06±10.38 μm in the blank control group, 499.86±34.00 μm in the TGF-β1 10 ng/ml group, 514.93±10.56 μm in the TNF-α 10 ng/ml group, 569.68±33.58 μm in the TGF-β1 10 ng/ml＋TNF-α10 ng/ml group. TGF-β1 cooperated with TNF-α led to scratch spacing narrowing significantly. Western blot analysis showed that expression of E-cadherin and Vimentin varied significantly in the TGF-β1＋TNF-α group. Conclusion Inducing human bronchial epithelial cell by TGF-β1 cooperated with TNF-α optimizes EMT model.

关 键 词：转化细胞生长因子-β1 肿瘤坏死因子-Α 上皮间质转化 

分 类 号：R56[医药卫生—呼吸系统]