Comparison of gel contraction mediated by airway smooth muscle cells from patients with and without asthma
Comparison of gel contraction mediated by airway smooth muscle cells from patients with and without asthma
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DOI：
10.1136/thx.2006.070474
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发表时间：
2007-10-01
期刊：
影响因子：
10
通讯作者：
McParland, Brent E.
中科院分区：
文献类型：
--
作者：
Matsumoto, Hisako;Moir, Lyn M.;McParland, Brent E.
背景：支气管过度收缩是哮喘患者对吸入性刺激最显著和危及生命的反应。然而，很少有研究调查这些患者气道平滑肌（ASM）的收缩性。本研究的目的是建立一种通过将ASM细胞包埋于胶原凝胶中来测量ASM细胞收缩的方法，并比较哮喘和非哮喘受试者之间的收缩情况。方法：将哮喘患者和非哮喘患者培养的ASM细胞置于铸型板上孵育过夜（方法1）或贴附（方法2），观察悬浮凝胶对组胺的凝胶收缩。同时检测平滑肌肌球蛋白轻链激酶蛋白水平。结果：含有ASM细胞的胶原凝胶在组胺刺激下呈浓度依赖性减小，在平均（SE）为15.7 (1.2)min时达到最大值。这种凝胶收缩被磷脂酶C （U73122）、肌球蛋白轻链激酶（ML-7）和Rho激酶（Y27632）抑制剂抑制。对比两组患者，方法1和方法2中含有哮喘患者ASM细胞的凝胶最大减少面积分别为19 (2)% （n = 8）和22 (3)% (n = 6)，均大于非哮喘患者的13 (2)% （n = 9, p = 0.05）和10 (4)% （n = 5, p = 0.024）。两组间平滑肌肌球蛋白轻链激酶水平无显著差异。结论：哮喘时ASM细胞收缩增强可能是哮喘支气管收缩过度的原因。
Backgrounds: Exaggerated bronchial constriction is the most significant and life threatening response of patients with asthma to inhaled stimuli. However, few studies have investigated the contractility of airway smooth muscle (ASM) from these patients. The purpose of this study was to establish a method to measure contraction of ASM cells by embedding them into a collagen gel, and to compare the contraction between subjects with and without asthma.Methods: Gel contraction to histamine was examined in floating gels containing cultured ASM cells from subjects with and without asthma following overnight incubation while unattached (method 1) or attached (method 2) to casting plates. Smooth muscle myosin light chain kinase protein levels were also examined.Results: Collagen gels containing ASM cells reduced in size when stimulated with histamine in a concentration-dependent manner and reached a maximum at a mean (SE) of 15.7 (1.2) min. This gel contraction was decreased by inhibitors for phospholipase C (U73122), myosin light chain kinase (ML-7) and Rho kinase (Y27632). When comparing the two patient groups, the maximal decreased area of gels containing ASM cells from patients with asthma was 19 (2)% (n = 8) using method 1 and 22 (3)% (n = 6) using method 2, both of which were greater than that of cells from patients without asthma: 13 (2)% (n = 9, p = 0.05) and 10 (4)% (n = 5, p = 0.024), respectively. Smooth muscle myosin light chain kinase levels were not different between the two groups.Conclusion: The increased contraction of asthmatic ASM cells may be responsible for exaggerated bronchial constriction in asthma.