草乌甲素对哮喘小鼠治疗作用及其机制研究  

Study on therapeutic effect and mechanism of BLA in asthmatic mice

作　　者：宋乔文 许月野 郭伟[1] 湛孝东[1] 赵金红[1] 李媛媛 徐静云 唐小牛[1,2] SONG Qiaowen;XU Yueye;GUO Wei;ZHAN Xiaodong;ZHAO Jinhong;LI Yuanyuan;XU Jingyun;TANG Xiaoniu(Department of Medical Parasitology,Wannan Medical College,Wuhu 241002,Anhui,China;Anhui Province Key Laboratory of Biological Macromlecules Research,Wannan Medical College)

机构地区：[1]皖南医学院医学寄生虫学教研室,安徽芜湖241002 [2]皖南医学院活性生物大分子研究安徽省重点实验室

出　　处：《中国病原生物学杂志》2024年第4期441-445,449,共6页Journal of Pathogen Biology

基　　金：安徽省高校自然科学重点基金(No.KJ2018A0263);一种植物来源的具有除螨去屑功效的洗发水研发(No.H202106)。

摘　　要：目的研究草乌甲素对哮喘小鼠的治疗作用及其对气道上皮紧密连接蛋白occludin,claudins以及黏蛋白5AC的影响。方法48只SPF级BALB/c雌鼠以每组12只随机分成4组,阴性对照组(NC组),哮喘组(AS组),草乌甲素(BLA组)和地塞米松阳性对照组(Dex组)。以粉尘螨提取浸液构建哮喘模型。酶联吸附反应(ELISA)检测血清IgE和肺泡灌洗液(BALF)中IL-4、IFN-γ及黏蛋白5AC(MUC5AC)的含量;苏木精-伊红(HE)和过碘酸-雪夫(PAS)染色观察肺组织病理改变;Western blot和免疫荧光检测肺组织气道上皮紧密连接蛋白occludin和claudins表达;RT-qPCR检测肺组织MUC5AC mRNA的表达。结果ELISA结果表明:与AS组(306.70±46.73)ng/mL相比,BLA组小鼠IgE(142.17±25.66)ng/mL水平显著下降;与AS组(242.35±41.81)pg/mL相比,BLA组小鼠IL-4(166.76±26.32)pg/mL水平显著下降;与AS组(272.34±57.32)pg/mL相比,BLA组小鼠IFN-γ(369.13±51.41)pg/mL水平显著上升;与AS组(1.42±0.34)ng/mL相比,BLA组小鼠MUC5AC(0.66±0.24)ng/mL水平显著下降(均P<0.01)。HE和PAS染色结果表明:与AS组相比,BLA组小鼠气道炎症细胞浸润,杯状细胞增生等病理改变明显缓解。Western blot结果表明:与AS组相比,BLA组小鼠occludin、claudin-1蛋白表达明显上调,claudin-4蛋白表达明显下调(均P<0.05)。免疫荧光结果表明:与AS组相比,BLA组小鼠气道上皮紧密性明显改善。RT-qPCR结果表明:与AS组相比,BLA组小鼠Muc5ac mRNA表达显著抑制(P<0.01)。结论草乌甲素可能通过调节气道上皮紧密连接蛋白和MUC5AC的表达水平改善哮喘小鼠气道上皮屏障功能。Objective To study the therapeutic effect of Bulleyaconitine A(BLA)on asthmatic mice and itseffects on occludin,claudins and Mucin5AC(MUC5AC).Methods 48female mice were randomly divided into 4groups:NC group,AS group,BLA group and Dex group,with 12mice in each group.The asthmatic model was established by the extract of Dermatophagoides farinae.Enzyme linked immunosorbent assay(ELISA)was used to detect the content of Immunoglobulin E(IgE)in serum and interleukin-4(IL-4),interferon-γ(IFN-γ)andMUC5AC in Bronchoalveolar lavage fluid(BALF).Hematoxylin-eosin(HE)and Periodic Acid-Schiff(PAS)staining were used to observe the pathological changes of lung tissues.Western blot and immunofluorescence were used to detect the expression of occludin and claudins in lung tissues.RT-qPCR was used to detect the expression of Muc5ac mRNA in lung tissues.Results Compared with AS group,the levels of IgE in BLA group was significantly decreased(306.70±46.73ng/mL vs 142.17±25.66ng/mL,P<0.01).Compared with AS group,the levels of IL-4in BLA group was significantly decreased(242.35±41.81pg/mL vs 166.76±26.32pg/mL,P<0.01).Compared with AS group,the levels of IFN-γin BLA group was significantly increased(272.34±57.32pg/mL vs 369.13±51.41pg/mL,P<0.01).Compared with AS group,the levels of MUC5AC in BLA group was significantly decreased(1.42±0.34ng/mL vs 0.66±0.24ng/mL,P<0.01).Compared with the AS group,the airway inflammatory cell infiltration and goblet cell proliferation were significantly relieved in the BLA group.Compared with AS group,the expression of occludin and claudin-1proteins in BLA group were significantly up-regulated(P<0.05),and claudin-4protein expression in BLA group was significantly down-regulated(P<0.05).Immunofluorescence results showed that BLA could significantly improve the tightness of airway epithelium in asthmatic mice.Compared with AS group,the expression of Muc5ac mRNA in BLA group was significantly inhibited(P<0.01).Conclusion BLA may improveairway epithelial barrier function of asthmatic mice by regul

关 键 词：过敏性哮喘 草乌甲素 上皮屏障 紧密连接蛋白 MUC5AC 

分 类 号：R285.5[医药卫生—中药学]