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气道平滑肌细胞(airway smooth muscle cells,ASMCs)是引起哮喘患者气道收缩狭窄、呼吸阻力增加的主要效应细胞。ASMCs收缩效应检测是研究哮喘病理生理机制、评估或研发新的支气管舒张药物的重要实验依据。而常用的活体组织张力检测以及单层培养ASMCs显微镜形态观察等方法存在样品取材或测量误差等问题。该研究利用胶原蛋白凝胶构建气道平滑肌细胞的三维立体培养模型,将凝胶与培养孔壁分离,在不同的时间点记录细胞收缩力作用下凝胶面积的变化值,以此反映ASMCs的收缩效应。结果显示,0.1,1,10 mmol/L的乙酰胆碱(acetylcholine,Ach)刺激后,凝胶面积均显著缩小,随着剂量增加ASMCs的收缩反应增强。提前加入肌球蛋白ATP酶抑制剂BDM(butanedione monoxime),能明显抑制Ach诱导的ASMCs收缩反应,说明该方法的可重复性和准确性好。
Airway smooth muscle cells (ASMCs) are the main effector cells that cause airway constriction and respiratory resistance in asthmatic patients. ASMCs systolic effect test is to study the pathophysiology of asthma, an important experimental evidence to evaluate or develop new bronchodilator drugs. However, there are some problems such as sample preparation or measurement error, which are commonly used in the detection of living tissue tension and microscopic morphology observation of monolayer cultured ASMCs. In this study, collagen gel was used to construct a three-dimensional culture model of airway smooth muscle cells, and the gel was separated from the culture hole wall. The change of gel area under cell contraction force was recorded at different time points to reflect the change of ASMCs Shrinkage effect. The results showed that after 0.1, 1, 10 mmol / L acetylcholine (Ach) stimulation, the gel area was significantly reduced, as the dose increased ASMCs contractile response increased. The addition of myosin ATPase inhibitor BDM (butanedione monoxime) could significantly inhibit Ach-induced ASMCs contractile response, indicating that the method is reproducible and accurate.