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支气管哮喘因严重影响病人的生活质量及远期预后,已成为呼吸系统常见的慢性疾病之一[1]。尤其是儿童病人,因临床表现及发作形式多样化,且患儿就诊时对于病史及症状描述不确切,更是增加了诊断的难度,导致诊治不及时,随着病程的迁延,患儿出现进行性加重的气道狭窄和不可逆性的气道重塑。因此,早期诊断和治疗儿童支气管哮喘的重要性不言而喻。促使哮喘向慢性疾病演变的重要因素在近年来的研究中被发现,主要是由于气道炎症及气道重塑,它们在哮喘发展过程中起着重要的病理生理作用[2]。有研究[3]表明,卡介苗(bacillus calmette-guerin, BCG)通过免疫调节,发挥其减轻气道上皮细胞损伤及间质转化的作用,从而减轻哮喘性气道炎症和气道阻塞,但目前国内外对BCG在改善气道重塑的作用机制方面进行报道的相关文献较少。本研究着重探讨在BCG干预下,支气管哮喘大鼠气道重塑情况以及血清缺氧诱导因子-1α(hypoxia-inducible factor-1 alpha,HIF-1α)的表达情况,旨在为临床控制哮喘症状、改善哮喘病人预后提供实验室依据,现作报道。
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通过光学显微镜进行观察,对照组大鼠肺组织肺泡形态正常,排列均匀,肺泡壁未见明显增厚,气管及血管结构均正常;哮喘组大鼠肺组织肺泡壁有明显增厚,视野可见大量炎性细胞浸润,气管腔内可见明显黏液残留,模型构建成功;BCG治疗组大鼠肺组织整体结构趋于正常,表明BCG对大鼠哮喘有治疗作用(见图 1)。
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3组大鼠血清HIF-1α表达水平比较,哮喘组> BCG治疗组>对照组,差异有统计学意义(P < 0.01)(见表 1)。
分组 n HIF-1α 对照组 10 10.12±0.95 哮喘组 10 20.85±2.47** BCG组 10 13.57±2.11**△△ F — 78.58 P — < 0.01 MS组内 — 3.819 q检验:与对照组比较**P < 0.01;与哮喘组比较△△P < 0.01 表 1 3组大鼠血清HIF-1α表达水平比较(x±s; pg/mL)
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与对照组比较,哮喘组和BCG治疗组大鼠WAt/Pi均升高(P < 0.01和P < 0.05),哮喘组大鼠WAm/Pi升高(P < 0.01);与哮喘组比较,BCG治疗组大鼠WAt/Pi和WAm/Pi均降低(P < 0.01和P < 0.05)(见表 2)。
分组 n WAt/ Pi WAm/ Pi 对照组 10 5.64±3.47 1.27±0.96 哮喘组 10 56.76±29.93** 3.68±2.33** BCG治疗组 10 28.08±11.90*△△ 1.79±0.63△ F — 17.99 6.55 P — < 0.01 < 0.01 MS组内 — 349.819 2.249 q检验:与对照组比较*P < 0.05,**P < 0.01;与哮喘组比较△P < 0.05,△△P < 0.01 表 2 3组大鼠WAt/Pi和WAm/Pi的比较(x±s; μm)
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结果显示,大鼠血清HIF-1α与支气管壁厚度和支气管平滑肌厚度之间呈正相关(r=0.618、0.503,P < 0.01)。
卡介苗对支气管哮喘大鼠气道重塑及血清HIF-1α表达的影响
Effect of BCG on the airway remodeling and serum level of HIF-1α in bronchial asthma rats
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摘要:
目的探讨卡介苗(BCG)干预下,支气管哮喘大鼠气道重塑以及血清缺氧诱导因子-1α(HIF-1α)的表达情况。 方法将30只清洁级健康雄性Sprague-Dawley(SD)大鼠随机分为对照组、哮喘组和BCG治疗组,每组10只。哮喘组大鼠采用腹腔注射10%卵白蛋白(OVA)+氢氧化铝混合液致敏,再用1% OVA超声雾化吸入的方法进行激发制作支气管哮喘模型;对照组大鼠采用相同的制模方法,但在相应的时间点使用0.9%氯化钠溶液替代致敏剂,进行腹腔注射和超声雾化吸入处理。BCG治疗组大鼠制模方法与哮喘组相同,每天雾化前半小时皮内注射BCG 0.025 mg进行干预治疗。3组大鼠在末次激发24 h后收集血清标本,采用ELISA法检测HIF-1α含量,采血后处死大鼠并制作气管和肺组织病理切片,观察大鼠气道炎症及气管重塑情况。运用图像分析软件测量支气管内径周长(Pi)、支气管管壁面积(WAt)、支气管平滑肌总面积(WAm)等指标, 并与Pi比较进行标化。 结果对照组大鼠肺组织中肺泡、气管、血管结构均正常;哮喘组大鼠肺泡壁明显增厚,气管腔可见黏液残留,部分可见肺泡腔内出血;BCG治疗组大鼠肺组织整体结构趋于正常。哮喘组大鼠血清中HIF-1α含量较对照组升高,经BCG干预后,大鼠血清HIF-1α降低(P < 0.01)。HIF-1α和支气管壁厚度(WAt/Pi)之间的相关性呈正相关,和支气管平滑肌厚度(WAm/Pi)之间的相关性呈正相关(P < 0.01)。 结论支气管哮喘大鼠经过BCG干预治疗,气道重塑减轻,可能是通过抑制HIF-1α的表达来发挥作用。 Abstract:ObjectiveTo investigate the effects of BCG intervention on the airway remodeling and serum hypoxia-inducible factor-1 alpha(HIF-1α)expression in bronchial asthma rats. MethodsThirtySD rats were randomly divided into the control group, asthma group and BCG treatment group(10 rats each group).The asthma group were intraperitoneally injected with 10% egg albumin(OVA)combined with aluminum hydroxide mixture for sensitization, and then stimulated by 1% OVA ultrasonic atomization inhalation to establish bronchial asthma model.The control group were treated with the same establishing model method, but at the corresponding time point, 0.9% sodium chloride solution was used to replace the sensitizer, and intraperitoneal injection and ultrasonic atomization inhalation were performed.The modeling method in BCG treatment group were the same as that of the asthma group, and the intradermal injection of 0.025 mg of BCG before half an hour of atomization was performed daily for intervention treatment.The inter-group variables were analyzed using Pearson correlation analysis method.The serum samples were collected after 24 h of last stimulation in three groups, and the contents of HIF-1α were detected using ELISA.After the rats were sacrificed, the trachea and lung histopathology sections were made to observe the airway inflammation and tracheal remodeling.The image analysis software was used to measure the internal diameter circumference(Pi), bronchial tube wall area(WAt)and total area of bronchial smooth muscle(WAm). ResultsThe structures of alveoli, trachea and blood vessels in control group were normal.In the asthma group, the alveolar wall significantly thickened, mucus remained in the trachea cavity, and intra-alveolar hemorrhage was partially observed.The overall structure of lung tissues in BCG treatment group tended to be normal.The serum HIF-1α content in asthma group increased compared with the control group, and the serum HIF-1 content decreased after BCG intervention(P < 0.01).The HIF-1α was positively correlated with bronchial wall thickness(WAt/Pi)and bronchial smooth muscle thickness(WAm/Pi)(P < 0.01). ConclusionsAfter BCG intervention treatment, the airway remodeling alleviates in bronchial asthma rats, and the effects may be by inhibiting HIF-1α expression to play. -
Key words:
- bronchial asthma /
- airway remodeling /
- Bacillus Calmette Guerin /
- hypoxia-inducible factor-1 alpha /
- rat
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表 1 3组大鼠血清HIF-1α表达水平比较(x±s; pg/mL)
分组 n HIF-1α 对照组 10 10.12±0.95 哮喘组 10 20.85±2.47** BCG组 10 13.57±2.11**△△ F — 78.58 P — < 0.01 MS组内 — 3.819 q检验:与对照组比较**P < 0.01;与哮喘组比较△△P < 0.01 表 2 3组大鼠WAt/Pi和WAm/Pi的比较(x±s; μm)
分组 n WAt/ Pi WAm/ Pi 对照组 10 5.64±3.47 1.27±0.96 哮喘组 10 56.76±29.93** 3.68±2.33** BCG治疗组 10 28.08±11.90*△△ 1.79±0.63△ F — 17.99 6.55 P — < 0.01 < 0.01 MS组内 — 349.819 2.249 q检验:与对照组比较*P < 0.05,**P < 0.01;与哮喘组比较△P < 0.05,△△P < 0.01 -
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