神经激肽-1受体拮抗剂WIN62577对哮喘气道重塑大鼠气管平滑肌细胞迁移的影响

Effects of NK-1R inhibitor WIN62577 on the migration of airway smooth muscle cells in asthmatic rats with airway remodeling

摘要:

目的 探讨哮喘气道重塑大鼠中气管平滑肌细胞迁移的变化及神经激肽-1受体(NK-1R)拮抗剂WIN62577对其的影响。方法 将Sprague-Dawley(SD)大鼠随机分为对照组和哮喘气道重塑组,应用卵清蛋白对模型组大鼠激发8周,原代培养纯化气管平滑肌细胞(ASMC),通过免疫荧光、实时定量PCR等方法,检测ASMC中NK-1R的表达,并给予不同剂量WIN62577干预,应用transwell小室检测ASMC迁移的变化。结果 哮喘气道重塑组ASMC中NK-1R主要分布在细胞质和细胞膜,其mRNA的表达高于对照组(P<0.01)。哮喘气道重塑组ASMC迁移数目较对照组明显增高(P<0.01),而分别给予不同浓度(10^-11 mol/L、10^-10 mol/L、10^-9 mol/L、10^-8 mol/L)的WIN62577干预后ASMC迁移数目较哮喘气道重塑组明显减少(P<0.05)。结论 NK-1R能通过促进ASMC迁移能力而影响哮喘气道重塑。

Abstract:

Objective To study the changes in the migration of airway smooth muscle cells (ASMC) in asthmatic rats with airway remodeling and the effect of NK-1R inhibitor WIN62577 on the migration of ASMC. Methods Sprague-Dawley rats were randomly assigned into two groups: airway remodeling induced by asthma and normal control. ASMC from rats with asthma and airway remodeling induced by ovalbumin (OVA) inhalation for 8 weeks were primary cultured and purified. Immunofluorescence and real-time PCR were used to measure the expression of NK-1R. With NK-1R inhibitor WIN62577 treatment, the changes in the migration of ASMC were measured by transwell chambers. Results NK-1R in ASMC was expressed mainly in the cytoplasm and cell membrane in the airway remodeling group, and the mRNA expression of NK-1R was higher than the normal control group (P<0.01). The number of the migrated ASMC in the airway remodeling group was significantly higher than that in the normal control group (P<0.01). Various concentrations (10^-11 mol/L, 10^-10 mol/L, 10^-9 mol/L and 10^-8 mol/L) of WIN62577 treatment decreased the number of the migrated ASMC (P<0.05). Conclusions NK-1R may affect airway remodeling possibly through promoting the migration ability of ASMC in rats with asthma.