维拉帕米对哮喘支气管杯状细胞的影响：致敏动物的实验研究。

Effect of verapamil on bronchial goblet cells of asthma: an experimental study on sensitized animals.

机构信息

Zakaria Research Center, Medical School of Islamic Azad University-Mashhad Branch, Mashhad, Iran.

出版信息

Pulm Pharmacol Ther. 2012 Apr;25(2):163-8. doi: 10.1016/j.pupt.2011.11.001. Epub 2011 Nov 25.

DOI:10.1016/j.pupt.2011.11.001

PMID:22133887

Abstract

INTRODUCTION

Goblet cell hyperplasia (GCH) and mucus hypersecretion in the airway is recognized as an important contributor to morbidity and mortality in asthma and COPD. Verapamil is a calcium channel blocker that binds to the alpha-subunit of L-type calcium channels and inhibits the mucin gene via the calmodulin and CaM kinase pathway. The objective of this study was to determine the in vivo effect of verapamil on GCH and eosinophilic inflammation in sensitized mice.

METHODS

Male BALB/c mice were sensitized to ovalbumin using the standard method. Two groups of animals were received verapamil via an intramuscular injection: 1-low dose (0.5 mg/kg/day for two weeks), 2-high dose (1.5 mg/kg/day for two weeks). Serum and bronchoalveolar lavage fluid (BALF) was collected and analyzed for inflammatory cells, interferon-γ and IL-4. The left lung was sent for histopathological evaluation, especially for periodic acid-Schiff (PAS), to identify goblet cells in the epithelium. The degree of inflammatory cell infiltration, including eosinophils, mucus plugging, and smooth muscle thickness of the airways were classified on a semi quantitative scale.

RESULTS

Inflammatory cell infiltration in peribronchial and perivascular areas was observed in all sensitized groups. Eosinophils percentage in the BALF significantly decreased in verapamil-treated mice compared with sensitized mice (from 19.8% in asthmatic to 5.4% for low dose and 4.4% for high dose). The ratio of airway goblet cells per epithelial cells were significantly lower in verapamil-treated mice versus sensitized mice (1.57±1.30% for low dose; 1.50±0.93% for high dose versus 12.93±7.55%, P<0.05, respectively). Mucus production of goblet cells decreased significantly in verapamil-treated mice versus sensitized mice (mean score was 1.45±0.30 for low dose; 0.81±1.00 for high dose versus 2.85±0.86 in the sensitized control group, P<0.05, respectively). The concentration of serum and BALF-IFN-γ in verapamil-treated mice markedly increased by the verapamil treatment when compared to sensitized mice (15.1±0.43 versus 4.7±0.96, P<0.05 and 91.8±47.7 versus 14.8±4.6, P<0.01, respectively).

CONCLUSION

Verapamil is a useful drug with therapeutic targeting on GCH and a potential way to limit mucous production and improve bronchial inflammation.

摘要

简介

杯状细胞增生（GCH）和气道黏液分泌过度被认为是哮喘和 COPD 发病率和死亡率的重要原因。维拉帕米是一种钙通道阻滞剂，它与 L 型钙通道的α亚单位结合，并通过钙调蛋白和 CaM 激酶途径抑制粘蛋白基因。本研究的目的是确定维拉帕米对致敏小鼠 GCH 和嗜酸性粒细胞炎症的体内作用。

方法

雄性 BALB/c 小鼠采用标准方法致敏卵白蛋白。两组动物通过肌肉注射接受维拉帕米：1-低剂量（0.5mg/kg/天，两周），2-高剂量（1.5mg/kg/天，两周）。收集血清和支气管肺泡灌洗液（BALF），分析炎症细胞、干扰素-γ 和白细胞介素-4。左肺用于组织病理学评估，特别是过碘酸-Schiff（PAS），以识别上皮中的杯状细胞。炎症细胞浸润的程度，包括嗜酸性粒细胞、黏液栓和气道平滑肌厚度，采用半定量评分进行分类。

结果

所有致敏组均观察到支气管周围和血管周围区域的炎症细胞浸润。与致敏小鼠相比，维拉帕米治疗小鼠的 BALF 中嗜酸性粒细胞百分比显著降低（从哮喘中的 19.8%降至低剂量的 5.4%和高剂量的 4.4%）。与致敏小鼠相比，维拉帕米治疗小鼠的气道杯状细胞与上皮细胞的比例显著降低（低剂量组为 1.57±1.30%；高剂量组为 1.50±0.93%，分别为 12.93±7.55%，P<0.05）。与致敏小鼠相比，维拉帕米治疗小鼠的杯状细胞黏液生成显著减少（低剂量组平均评分 1.45±0.30；高剂量组 0.81±1.00，致敏对照组 2.85±0.86，P<0.05）。与致敏小鼠相比，维拉帕米治疗小鼠的血清和 BALF-IFN-γ 浓度在维拉帕米治疗后显著增加（15.1±0.43 与 4.7±0.96，P<0.05 和 91.8±47.7 与 14.8±4.6，P<0.01）。