基质黏附强度调控血小板生长因子诱导的大鼠气道平滑肌细胞中Fyn激酶活性  

Fibronectin adhesion strength regulates PDGF-induced Fyn activity in rat airway smooth muscle cells

作　　者：姚辉 邓林红 欧阳明星 YAO Hui;DENG Linhong;OUYANG Mingxing(Institute of Biomedical Engineering and Health Sciences,Changzhou University,Changzhou 213164,China)

机构地区：[1]常州大学生物医学工程与健康科学研究院

出　　处：《生物医学工程研究》2019年第4期381-386,共6页Journal Of Biomedical Engineering Research

基　　金：国家自然科学基金资助项目(11872129、11532003、31670950);江苏省科技厅资助项目(BK20181464);常州市科技项目(CZ20180017)

摘　　要：气道平滑肌(airway smooth muscle,ASM)有收缩和舒张的功能,而哮喘的气道高反应中,ASM保持高收缩状态的表型,引起支气管痉挛和呼吸功能受损,但其生物力学机制尚不清楚。Fyn作为非受体酪氨酸激酶Src家族成员,在多个生理、病理过程中起作用。炎症因子如血小板生长因子(platelet-derived growth factor,PDGF)能刺激ASM细胞的增殖和诱导收缩延长效应,利用荧光共振能量转移技术(fluorescence resonance energy transfer,FRET),本研究发现PDGF能快速高效地激活ASM细胞中Fyn的活性(约110%FRET变化)。通过改变表面Fibronectin的包被浓度(1.25~400μg/mL)调节细胞-基质黏附强度,证实PDGF诱导Fyn的激活水平与黏附强度有正相关性,显示力学微环境影响ASM细胞中化学信号的可能性。研究中,减弱细胞内部收缩力对Fyn激活水平无明显影响。此初步性工作对理解哮喘条件下力学微环境改变对ASM细胞内的生化信号影响提供了一些思路。Airway smooth muscle(ASM)in respiratory system has regular contraction and relaxation,whereas during airway hyper-responsiveness under asthma condition,ASM maintains contraction force to prevent air flow through bronchial tubes.Fyn as one member of non-receptor tyrosine kinase Src family is involved into multiple physiological and pathological processes,and reportedly fibronectin-coated substrate rigidity-regulated biomechanical response.Here we tried to look at Fyn activity by fluorescence resonance energy transfer(FRET)technology in cultured rat ASM cells.Platelets-derived growth factor(PDGF),which was reported able to cause ASM proliferation and hypercontractivity,induced a rapid and efficient Fyn activation measured by about 110% FRET change in ASM cells.By coating the cover slips with gradient fibronectin concentrations ranging from 1.25 to 40μg/mL,we found a positive correlation between PDGF-induced Fyn activity and cell-fibronectin adhesion strength,indicating a possible influence of mechanical microenvironment on biochemical signaling in ASM cells.Reduction of intracellular contraction force didn’t obviously change PDGF-induced Fyn activity.This work may help provide some clue to the possible impact on biochemical signals in ASM cells from changed mechanical microenvironment under asthma condition.

关 键 词：气道平滑肌细胞 Fyn激酶 细胞黏附强度 荧光共振能量转移 血小板生长因子 

分 类 号：R318[医药卫生—生物医学工程]