金莲花胶囊对卵清蛋白诱导的过敏性哮喘小鼠气道炎症的作用研究

Effects of Jinlianhua capsules on airway inflammation in ovalbumin-induced allergic asthma mice

为探讨金莲花胶囊对卵清蛋白诱导的过敏性哮喘小鼠气道炎症的作用,将C57BL/6J小鼠随机分为5组,分别为对照组、模型组、阳性药物地塞米松组(2 mg·kg^(-1))和金莲花胶囊给药组(200和400 mg·kg^(-1)),每组6只。采用卵清蛋白和铝佐剂致敏小鼠,每周1次,连续3周;气管滴注卵清蛋白激发1次,滴鼻法给予卵清蛋白激发4 d,建立哮喘模型。金莲花胶囊给药组小鼠D19开始灌胃给药,连续给药7 d;阳性药物组腹腔注射给药7 d。末次激发24 h(D26),检测肺重/体重比值,HE染色观察肺组织形态学变化,PAS染色观察气道上皮细胞黏液分泌状态,天狼星红染色观察肺组织嗜酸性粒细胞浸润,分光光度法检测肺组织嗜酸性粒细胞过氧化物酶活性。结果表明:与模型组相比,金莲花胶囊能降低哮喘小鼠肺重/体重比值,改善气道嗜酸性粒细胞炎症,减少黏液高分泌和PAS评分,降低肺组织嗜酸性粒细胞过氧化物酶活性;还能减少肺组织中IL-5含量,抑制肺组织IL-5、IL-13、Muc5ac和Eotaxin的mRNA水平。且金莲花胶囊(12.5~50μg·mL^(-1))可体外抑制卵清蛋白刺激的致敏小鼠脾淋巴细胞分泌IL-5。此外,金莲花胶囊400 mg·kg^(-1)组还能抑制哮喘小鼠肺组织p38 MAPK蛋白质磷酸化水平。综上,金莲花胶囊可能通过p38 MAPK途径抑制OVA诱导的过敏性哮喘小鼠气道炎症。

To investigate the effect of Jinlianhua capsules on airway inflammation in ovalbumin(OVA)-induced allergic asthma mice.C57BL/6 Jmice were randomly divided into 5 groups with 6 mice in each group,including control group,model group,dexamethasone(2mg·kg^(-1))group and Jinlianhua capsule groups(200 and 400mg·kg^(-1)).The mice were sensitized with OVA plus aluminum adjuvant once a week for 3 weeks,and the asthma model was established by transtracheal instillation and intranasal instillation of OVA for 5 days.Mice in Jinlianhua capsule groups were intragastrically administrated with Jinlianhua capsule from D19 to D25 for 7 days,and mice in positive drug group were injected intraperitoneally with dexamethasone for 7 days.24 hours after the last challenge,the ratio of lung weight to body weight was measured.The slice of lung tissue was conducted by HE staining,PAS staining and Sirius red staining for observing the morphological changes of lung tissue,the mucus secretion of airway epithelial cells and the eosinophil infiltration in lung tissue,respectively.The activity of eosinophil peroxidase in lung tissue was detected by spectrophotometry.The content of IL-5 in lung tissue homogenate and the content of IL-5 secreted by spleen cells of mice sensitized by OVA were detected by ELISA.The mRNA levels of IL-5,IL-13,Muc5ac and Eotaxinin lung tissue were detected by qRT-PCR.The phosphorylation level of p38 MAPK protein in lung tissue was detected by Western blot.The results showed that compared with the model group,the ratio of lung weight to body weight and the activity of peroxidase in lung tissue were reduced in Jinlianhua capsule groups.The results of morphological changes of lung tissue showed that Jinlianhua capsule reduced airway eosinophil inflammation and mucus hypersecretion.The content of IL-5 in lung tissue and the mRNA levels of IL-5,IL-13,Muc5ac and Eotaxinin lung tissue of asthmatic mice were decreased after Jinlianhua capsule treatment.Moreover,Jinlianhua capsule(12.5-50μg·mL^(-1))could also inhibit the secretion of IL-5 by spleen lymphocytes of sensitized mice stimulated by ovalbumin in vitro.In addition,the expression of p38MAPK protein phosphorylation in lung tissue of asthmatic mice was inhibited after 400mg·kg^(-1)Jinlianhua capsule treatment.In conclusion,Jinlianhua capsule alleviates airway inflammation in OVA-induced allergic asthma mice through inhibiting p38MAPK pathway.