机械牵拉对人支气管上皮样细胞气道重塑相关因子表达的影响  

Effects of Mechanical Stretch on Expression of Airway Remodeling Associated Factors in Human Bronchial Epithelioid Cells

作　　者：余倩[1] 李敏超[1] YU Qian LI Minchao(Department of Respiratory Medicine, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China)

机构地区：[1]重庆医科大学附属第二医院呼吸内科,重庆400010

出　　处：《生物医学工程学杂志》2016年第5期923-930,共8页Journal of Biomedical Engineering

基　　金：国家自然科学基金面上基金资助项目(81270102);重庆市自然科学基金资助项目(cstc2012jjA10050);重庆市教委科学技术研究资助项目(KJ120301)

摘　　要：本文为了研究机械牵拉人支气管上皮样细胞(16HBE)对转化生长因子-β1(TGF-β1)、成纤维细胞生长因子-2(FGF-2)表达的影响及其相关信号转导机制。采用柔性基底加载装置(FX-4000T)牵拉16HBE细胞,参数设置为拉伸幅度15%,频率1Hz,加载时间分别为:0.5h、1h、1.5h、2h。选择TGF-β1、FGF-2和Ca^(2+)表达较高的一组以经典瞬时感受器电位1(TRPC1)抑制剂SKF96365、蛋白激酶C(PKC)抑制剂HA-100干预。发现与空白对照组相比,4个时间段TGF-β1、FGF-2和Ca^(2+)的表达均高于空白对照组,差异有统计学意义(P<0.05),在0.5h时其增幅最大。对0.5h机械牵拉组予以SKF96365、HA-100干预,其TGF-β1、FGF-2和Ca^(2+)的表达均较0.5h机械牵拉组降低,差异有统计学意义(P<0.05)。本文研究结果说明,机械牵拉16HBE细胞可导致气道重塑相关因子表达增加,PKC、TRPC1、Ca^(2+)可能参与了该信号通路。This study is aimed to investigate the effects of mechanical stretch on the expression of transforming growth factor-β1 （TGF-β1） and fibroblast growth factor-2 （FGF-2）, and the signaling pathway in human bronchial epithelioid （16HBE） cells under mechanical stretch. Using loading device with flexible substrate （FX-4000T） to stretch 16HBE cells, we found that the stretching elongation was 15%, at frequency of 1 Hz, stretching for 0.5 h, 1 h, 1.5 h and 2 h. Choosing the higher expression of TGF-β1, FGF-2 and Ca^2＋ group to carry out intervention experiments, we used the cells pretreated with canonical transient receptor potential 1 （TRPC1） channel antagonist SKF96365, protein kinase C （PKC） inhibitor HA-100, and thereafter mechanical stretch to interpose. Compared with those in the blank control group, TGF-β1 and FGF-2＇ protein and mRNA, intracellular Ca^2＋ fluorescence intensity were higher, and the differences were statistically significant （P 〈 0.05） at the 4 time points, 0.5 h, 1 h, 1.5 h and 2 h. At 0.5 h, the increasing rate was the highest. TGF-β1 protein and mRNA, FGF-2 protein and mRNA, intracellular Ca^2＋ luorescence intensity in the stretch＋SKF96365 and stretch＋HA-100 intervented group were decreased, the differences were statistically significant than those in 0.5 h stretch group （P 〈 0.05） without intervention. The expression of TGF-β1, FGF-2 was up-regulated in 16HBE cells under mechanical stretch, PKC, TRPC1, and Ca^2＋ may participate in the signal path.

关 键 词：机械牵拉 人支气管上皮样细胞 转化生长因子-Β1 成纤维细胞生长因子2 

分 类 号：R56[医药卫生—呼吸系统]