首页|鼻敏方对变应性鼻炎肺脾气虚证模型大鼠鼻黏膜TMEM16A/NF-κB/MUC5AC信号通路的影响

鼻敏方对变应性鼻炎肺脾气虚证模型大鼠鼻黏膜TMEM16A/NF-κB/MUC5AC信号通路的影响

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目的 探讨鼻敏方治疗变应性鼻炎(AR)肺脾气虚证黏液高分泌的可能作用机制.方法 34只SD大鼠随机分为空白组(8只)、模型组(8只)、鼻敏方低剂量组(8只)、鼻敏方高剂量组(10只).除空白组外,其余各组大鼠以烟熏+番泻叶灌胃+卵清蛋白法建立AR肺脾气虚证大鼠模型.造模结束后,鼻敏方低、高剂量组大鼠分别给予鼻敏方浓缩液(浓度为2.16g/ml)1.08g/100g、2.16g/100g灌胃,模型组给予0.5 ml/100 g生理盐水灌胃,均每日1次,持续28天;空白组不干预.干预后进行行为学评价,ELISA法检测大鼠外周血总免疫球蛋白E(IgE)水平,HE染色观察大鼠鼻上皮黏膜病理学变化,免疫组化法检测鼻上皮黏膜的跨膜蛋白16A(TMEM16A)、黏蛋白5AC(MUC5AC)蛋白表达,Western Blot法检测鼻上皮黏膜核转录因子κB(NF-κB)蛋白表达,RT-PCR法检测鼻上皮黏膜TMEM16A、MUC5AC、NF-κBmRNA表达.结果 HE染色显示,空白组大鼠鼻上皮黏膜上皮细胞结构完整无脱落,无水肿、坏死等病变;模型组大鼠鼻上皮黏膜上皮组织增厚且部分脱落,可见嗜酸性粒细胞和淋巴细胞浸润;鼻敏方高、低剂量组大鼠鼻上皮黏膜组织病理均有改善,且鼻敏方高剂量组改善更明显.与空白组比较,模型组大鼠行为学积分及外周血总IgE水平显著升高,鼻上皮黏膜TMEM16A、MUC5AC、NF-κB蛋白及mRNA表达升高(P<0.05或P<0.01).与模型组比较,鼻敏方高剂量组大鼠行为学积分及外周血总IgE水平降低,鼻上皮黏膜TMEM16A、MUC5AC、NF-κB蛋白及mRNA表达降低(P<0.05或P<0.01);鼻敏方低剂量组大鼠行为学积分及外周血总IgE水平降低,鼻上皮黏膜TMEM16A、MUC5AC蛋白及mRNA表达降低,NF-κB蛋白表达降低(P<0.05或P<0.01),NF-κB mRNA表达差异无统计学意义(P>0.05).与鼻敏方低剂量组比较,鼻敏方高剂量组NF-κB蛋白表达降低(P<0.01).结论 鼻敏方可能通过调控鼻上皮黏膜TMEM16A/NF-KB/MUC5AC信号通路改善AR肺脾气虚症状及黏液高分泌.
Effects of Bimin Formula(鼻敏方)on the Nasal Mucosa TMEM16A/NF-κB/MUC5AC Signaling Pathway in a Rat Model of Allergic Rhinitis with Lung-Spleen Qi Deficiency
Objective To explore the possible mechanism of Bimin Formula(鼻敏方)in treating lung-spleen qi deficiency syndrome of allergic rhinitis(AR)with high mucin secretion.Methods Thirty-four SD rats were randomly divided into a blank group(8 rats),a model group(8 rats),a low-dose Bimin Formula group(8 rats),and a high-dose Bimin Formula group(10 rats).Except for the blank group,the other groups were subjected to AR lung-spleen qi deficiency rat models induced by smoking,gavage of Ginkgo biloba leaf extract,and ovalbumin.After modeling,rats in the low-and high-dose Bimin Formula groups were given Bimin Formula concentrate(concentration of 2.16 g/ml)by gavage at doses of 1.08 g/100 g and 2.16 g/100 g,respectively,while rats in the model group were given 0.5 ml/100 g of normal saline by gavage,once daily for 28 days;the blank group was not intervened.Behavioral assessments were performed after intervention.ELISA was used to detect the levels of peripheral blood total immuno-globulin E(IgE).HE staining was used to observe the pathological changes of nasal mucosa epithelium in rats,while immunohistochemistry was used to detect the expression of transmembrane protein 16A(TMEM16A)and mucin 5 AC(MUC5AC)protein in nasal mucosa.Western Blot was used to detect the expression of nuclear factor kappa B(NF-κB)protein,and RT-PCR was used to detect the expression of TMEM16A,MUC5AC,and NF-κB mRNA in nasal mucosa.Results HE staining showed that the nasal mucosa epithelial cell structure in the blank group was intact without shedding,swelling,or necrosis;the nasal mucosa epithelial tissue of rats in the model group was thickened and par-tially shed,with infiltration of eosinophils and lymphocytes visible;the pathological changes in nasal mucosa tissue of rats in the high-and low-dose Bimin Formulagroups were improved,and more improvement was showen in the high-dose group.Compared with those in the blank group,the behavioral scores and peripheral blood total IgE levels of rats in the model group significantly increased,as well as the expression of TMEM16A,MUC5AC,and NF-κB pro-teins and mRNA in nasal mucosa(P<0.05 or P<0.01).Compared with those in the model group,the behavioral scores and peripheral blood total IgE levels of rats in the high-dose Bimin Formula group decreased,and the expres-sion of TMEM16A,MUC5AC,and NF-κB proteins and mRNA in nasal mucosaalso decreased(P<0.05 or P<0.01);the behavioral scores and peripheral blood total IgE levels of rats in the low-dose Bimin Formula group were reduced,and the expression of TMEM16A and MUC5AC proteins and mRNA in nasal mucosa,as well as the expres-sion of NF-κB protein decreased(P<0.05 or P<0.01),but the difference in NF-κB mRNA expression was not statistically significant(P>0.05).Compared with the low-dose Bimin Formula group,the expression of NF-κB pro-tein in the high-dose group decreased(P<0.01).Conclusion Bimin Formula may improve the symptoms and high mucus secretion of AR lung-spleen qi deficiency by regulating the TMEM16A/NF-κB/MUC5AC signaling path-way in nasalmucosa.

allergic rhinitislung-spleen qi deficiency syndromeBimin Formula(鼻敏方)transmembrane protein 16Anuclear factor kappa Bmucin 5AChigh mucin secretion

徐宁聪、画奕惟、谭希、王今晗、梁紫华、周世卿、李云英、陈文勇、夏纪严、罗秋兰

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广州中医药大学第二临床医学院,广东省广州市白云区机场路12号,510405

广州中医药大学第二附属医院

广州中医药大学中药学院

变应性鼻炎 肺脾气虚证 鼻敏方 跨膜蛋白16A 核转录因子κB 黏蛋白5AC 黏液高分泌

国家自然科学基金广州市科技局基础研究计划(2022)广东省基础与应用基础研究基金企业联合基金(公共卫生与医药健康领域)项目(2021)全国名老中医专家传承工作室建设项目广东省中医药局中医药科研项目(科研平台专项)(2023)广东省普通高等学校青年创新人才类项目(2021)广东省中医院中医药科学技术研究专项广东省中医院中医药科学技术研究专项

816036672022010204732021A1515220007粤中医办函[2022]52号202330182021KQNCX014YN2020MS10YN2018ML09

2024

中医杂志
中华中医药学会 中国中医科学院

中医杂志

CSTPCD北大核心
影响因子:1.464
ISSN:1001-1668
年,卷(期):2024.65(8)
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