摘要
目的:观察钙池操纵性钙通道抑制剂SKF96365对小鼠慢性哮喘模型气道重塑和气道高反应性的影响。方法:用鸡卵清白蛋白(OVA)致敏和激发小鼠,建立慢性哮喘模型,33只雌性BALB/c小鼠随机分为3组:对照组、哮喘组、SKF96365组,每组11只,其中SKF96365组于每次激发前30 min给予SKF96365(10mg/kg)干预。哮喘组和SKF96365组于第0、7、14 d腹腔注射(i.p)200μL致敏液(含OVA粉剂20μg、氢氧化铝凝胶2 mg);自第21天起,腹腔注射1%戊巴比妥钠(70mg戊巴比妥钠/kg小鼠体重)麻醉小鼠后,OVA(40μg)滴鼻(i.n),连续6周,每周3次,共18次。对照组则在相同时间给予相应剂量的生理盐水腹腔注射和滴鼻。最后一次激发后24 h,各组分别随机取5只小鼠用于检测组织病理学变化,另6只小鼠采用Buxco小动物肺功能仪检测气道高反应性。其中组织病理学检测计算杯状细胞(过碘酸希夫染色阳性,PAS+)、胶原细胞(Masson阳性)和平滑肌细胞(α-SMA阳性)阳染面积/支气管基底膜周长值来评估小鼠气道重塑;观察给予不同浓度乙酰甲胆碱雾化时的气道阻力(resistance index,RI)最大值,评估小鼠气道高反应性。结果:对照组未见PAS阳性染色区域,哮喘组和SKF96365组杯状细胞增生高于对照组(7.29±2.04,4.49±1.70 vs 0.00±0.00,均P<0.01),且SKF96365组低于哮喘组(P<0.05)。Masson染色显示哮喘组和SKF96365组上皮下胶原沉积高于对照组(9.23±1.41,7.30±1.33 vs 1.60±0.77,均P<0.01),且SKF96365组低于哮喘组(P<0.05)。α-SMA免疫组化显示哮喘组和SKF96365组平滑肌增生肥大高于对照组(4.54±1.05,3.15±0.57 vs 1.97±0.69,均P<0.05),且SKF96365组低于哮喘组(P<0.05)。当乙酰甲胆碱(Mch)≤6.25 mg/m L时,3组小鼠气道阻力无显著差异(P>0.05),当25 mg/m L>Mch≥12.25 mg/m L时,哮喘组小鼠气道阻力明显大于对照组小鼠(P<0.001),当Mch≥25 mg/m L时,哮喘组小鼠气道阻力大于SKF96365组(P<0.05)。结论:采用SKF96365干预后,慢性哮喘
Objective： To investigate the effects of store operated calcium channel blocker SKF96365 on airway remodeling and airway hyperresponsiveness in mice model of chronic asthma. Methods： Thirty-three female BALB/c mice were randomly divided into three groups（ 11 mice in each group） ： control group,asthma group and SKF96365 group. The animals in asthma group and SKF96365 group were sensitized and challenged with ovalbumin（ OVA） to establish asthma model,and those in control group were given normal saline. The mice in the SKF96365 group were intraperitoneally injected with SKF96365（ 10 mg/kg） 30 min before OVA challenge. The asthma group and SKF96365 group were intraperitoneally injected with 20 g OVA emulsified in 2 mg alum in a total volume of 200 on days 0,7 and 14. These mice were then challenged by intranasal administration with OVA（ 40） for 6 weeks,3 times per week after anesthesia. The control group received normal saline at the same time. After last challenge periodic acid-Schiff（ PAS） staining was performed to evaluate goblet cell hyperplasia,and Masson-trichrome staining to evaluate the deposition of collagen matrix. In addition,immunohistochemistry on the α-smooth muscle actin（ α-SMA） was applied to examine airway smooth muscle cell hyperplasia and hypertrophy. The positive staining with PAS,Masson,α-SMA areas vs homologous bronchial basement membrane perimeter（ μm^2/μm） was used to indicate the degree of airway remodeling. Airway resistance index（ RI） of expiratory phase with different doses of acetyl-β-methylcholine chloride（ Mch） was measured. Results： In the asthma group and the SKF96365 group,the degree of the goblet cell hyperplasia was significantly higher than that in the control group（ 7. 29 ± 2. 04,4. 49 ± 1. 70 vs 0. 00 ± 0. 00,P〈0. 01）,and the level of goblet cell hyperplasia in the SKF96365 group was lower than that in the asthma group（ P〈0. 05）. The Masson staining showed that the deposition of collagen in the asthma group and the
作者
曹欢
王译民
谢宝娟
朱芳芳
李瑞婷
杨炯
高亚东
CAO Huan;WANG Yi-min;XIE Bao-juan;ZHU Fang-fang;LI Rui-ting;YANG Jiong;GAO Ya- dong(Zhongnan Hospital of Wuhan University,Wuhan 430071,China)
出处
《内科急危重症杂志》
2018年第3期248-252,共5页
Journal of Critical Care In Internal Medicine
