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目的研究层黏蛋白(laminin,LN)、纤维连接蛋白(fibronectin,FN)、I型胶原(collagen I,Col I)3种不同细胞外基质(extracelluar matrix,ECM)蛋白对血小板生长因子(platelet derived growth factor-BB,PDGF-BB)诱导的气道平滑肌细胞(airway smooth muscle cells,ASMCs)形态及收缩变化的影响。方法将ASMCs分别附着在LN、ColI、FN表衬的细胞培养皿中,并分成2组,分别在有或无PDGF-BB(10 mg/L)的无血清培养基中培养0～5 d。然后采用显微图像方法观测细胞形态和宽长比,光学磁微粒扭转细胞测量方法测量细胞受KCl或组胺(histamine)刺激时的收缩反应。结果在有PDGF-BB的培养条件下,ASMCs形态总体上变细变长,即细胞宽长比减小,但附着在LN上的ASMCs比附着在Col I或FN上的细胞宽长比相对要大。在无PDGF-BB的培养条件下,ASMCs对KCl刺激的收缩响应随培养时间增加而增加,但不受ECM蛋白成分的影响。而在有PDGF-BB的培养条件下,ASMCs对KCl或Histamine刺激的收缩响应总体上随培养时间呈下降趋势,但附着在LN上的ASMCs收缩响应下降程度相对较小。结论 ASMCs受PDGF-BB作用时,其形态和收缩性的变化与不同ECM蛋白成分有关,相对于Col I和FN,附着在LN的细胞形态和收缩性的变化较小。ECM蛋白成分对PDGF-BB诱导的ASMCs形态和收缩性变化的差异性影响,对于深入认识基质蛋白、炎症因子与气道平滑肌细胞的相互作用及其与哮喘病理生理机制的关系具有重要意义。
Objective To investigate the effect of three different extracellular matrix proteins (laminin, fibronectin (FN) and collagen I (Col I) on platelet derived growth factor-BB, PDGF-BB) induced airway smooth muscle cells (ASMCs) morphology and contraction changes. Methods ASMCs were respectively attached to the cell culture dishes lined with LN, ColI, FN and divided into two groups and cultured for 0-5 days in serum-free medium with or without PDGF-BB (10 mg / L) respectively. Then the cell morphology and the ratio of length to length were observed by microscopic image method, and the contractile response of the cells stimulated by KCl or histamine was measured by optical magnetic particle twisting cell measurement method. Results In the presence of PDGF-BB, the morphological changes of ASMCs were generally lengthened, ie, the ratio of cell length to length was decreased. However, the length of ASMCs attached to LN was longer than that of cells attached to Col I or FN Big. In the absence of PDGF-BB, the response of ASMCs to KCl stimulation contracted with increasing incubation time, but was not affected by ECM protein content. However, in the presence of PDGF-BB, the response of ASMCs to KCl or Histamine stimulation decreased generally with incubation time, but the reduction in ASMCs’ attachment to LN was relatively small. Conclusions The changes of morphological and contractile changes of ASMCs induced by PDGF-BB are related to different ECM protein components. Compared with Col I and FN, changes of morphology and contractility of ASMCs adhered to LN are small. The differential effect of ECM protein components on morphological and contractile changes of ASMCs induced by PDGF-BB is of great significance for understanding the interaction between matrix proteins, inflammatory cytokines and airway smooth muscle cells and their relationship with the pathophysiology of asthma.