摘要
目的建立Balb/C小鼠的急性哮喘模型,应用复方黄龙汤进行治疗,并与地塞米松治疗效果比较,探讨复方黄龙汤对小鼠哮喘气道炎症的影响及作用机制。方法选取24只6～8周龄雌性Balb/C小鼠,体重(20±2)g,应用随机数字法随机分为正常对照组(A组)、哮喘模型组(B组)、地塞米松治疗组(C组)和复方黄龙汤治疗组(D组),每组6只。应用卵白蛋白致敏液腹腔注射联合OVA雾化吸入法建立哮喘模型,B、C、D组在第0、14天向小鼠腹腔注射OVA配置而成的致敏液0.2ml,第28、29、30天吸入1%OVA/PBS 10ml激发哮喘,造模23天后,C组每天给予地塞米松(2mg/kg),D组每天给予复方黄龙汤(2.7ml/kg);激发结束后,用无创肺功能测量仪测量小鼠气道反应性,qRT-PCR法测定小鼠肺组织中T-bet及GATA-3mRNA的含量,酶联吸附反应测定肺泡灌洗液中IL-4及IFN-γ含量,将肺泡灌洗液离心沉淀后涂片进行中性粒细胞、嗜酸性粒细胞和淋巴细胞等炎性细胞计数,用HE染色肺组织,评价病理改变。结果无创肺功能显示雾化吸入乙酰胆碱剂量>12.5μg/ml时,C组和D组penh值明显低于B组(P<0.05),但高于A组,差异无统计学意义;肺组织中T-bet mRNA表达B组高于A组(P<0.001),D组含量高于B组,但差异无统计学意义(P>0.05)。而GATA-3mRNA表达C、D组低于B组(均P<0.01)。肺泡灌洗液中IL-4含量B组高于A、C和D组(均P<0.01),而IFN-γ含量B组远低于A组(均P<0.01),D组肺泡灌洗液中IFN-γ含量较B组升高,差异有统计学意义(P<0.05);在小鼠肺泡灌洗液中B组嗜酸粒细胞(EOS)比C、D组明显增多(P<0.01);B组气道壁和肺组织中大量炎性细胞浸润(以嗜酸性粒细胞、淋巴细胞为主),支气管管壁增厚,管腔狭窄,与A组比较差异明显;C、D两组小鼠细支气管管腔呈圆形,较规整,其中C组管壁增厚较D组明显,气道壁和肺组织可见嗜酸粒细胞(EOS)及少量淋巴细胞浸润,基底膜无增厚,与B组比较明显减轻。结论复方黄龙汤可通过降低小鼠肺组织炎症反应,减轻支气管重塑,改善肺功能,其作用可能是通过上调T-bet和下调GATA-3平衡来实现的。
Objective To investigate the effect and mechanism of Fu-Fang- Huang- Long Decoction on acute airway inflammation of asthmatic mice. Methods Health female Balb/C mice were randomly divided into four groups： control group （group A）, asthma group （group B）, dexamethasone group （group C） and Fu-Fang-Huang- Long Decoction group （group D）. Asthma rat model was induced by ovalbumin （OVA, via intraperitoneal injection） combined with OVA aerosol administration. After modeling, dexamethasone （2mg/kg） and Fu Fang Huang Long decoction （2.7ml/kg） was administra- ted in group C and D, respectively. The airway reactivity （AR） was determined with pulmonary function test apparatus. The expression of T-bet/GATA-3mRNA was determined using q-RT-PCR. The level of IL-4 and IFN-7 in alveolar ]avage fluid was evaluated using enzyme linked immunosorbent assay （ELISA）. Hematoxylin and Eosin （HE） staining was per- formed to evaluate the pulmonary pathologies. Results None -invasive lung ruction reveals that when chloride acetycholine dose is higher than 12.5μg/ml, the penh value in groups C and D is significantly lower than group B（P〈0. 05）, but higher than group A. There is no significant difference between group C and D. Significant decrease of T-bet mRNA expression was noted in group B compared with group A（P〈0. 001）. Though T-bet mRNA expression in group D is higher than group C, there is no significant difference between them（P〉0.05）. For the expression of GATA-3 mR.NA, remarkable increase was noted in group B compared with group C and D. Higher expression of IL-4 was noted in group B compared with the other groups （P〈0.01）. For the level of IFN-7, remarkable decrease was noted in group B compared with group A. Significant increase was noted in BALF, eosinophitic granulocytes （EOS） in group B compared with those obtained from group C and D, respectively （P^0.01）. A large number of inflammatory cells were in the airway wall and lung tissue of Group B （eo- sinophils, lymphocyte predominant）. There is significant differences compared with group A ,such as the bronchial wail in group B become thicker and luminal narrower; The small bronchial lumen in group C and D is round and completely. The airway wall and lung tissue in group C showed eosinophils and a small amount of lymphocytic infiltrate basement membrane, which is significantly reduced in group B. Conclusions Fu-Fang- Huang- Long Decoction could relieve the pulmonary in- flammatory reaction and improve pulmonary function, which may resulted from the two-way regulation of T-bet and GATA- 3mRNA balance.
出处
《西部医学》
2012年第11期2051-2056,共6页
Medical Journal of West China
基金
国家自然科学基金资助项目(编号:30600265)
关键词
哮喘
气道炎症
转录因子
复方黄龙汤
地塞米松
Asthma
Airway inflammation
Transcription factors
Fu-Fang- Huang-Long decoction
Dexamethasone
