炎症微生态响应TAS2R14/SIgA/TSLP通过肺-肠轴以射干麻黄汤及苦泄方调控寒哮大鼠上皮细胞屏障的机制研究  

Mechanism of inflammatory microecological response to TAS2R14/SIgA/TSLP in regulating epithelial cell barrier in cold asthma rats through lung-gut axis by using Shegan Mahuang Decoction and bitter and purging Chinese herbs

作　　者：袁亚美[1,2,3] 叶卫东 程悦 李秋慧 刘家鑫[1] 乔佳乐 王坤 方向明[1,2,3] YUAN Ya-meil;YE Wei-donggl;CHENG Yue;LI Qiu-hui;LIU Jia-xin;QIAO Jia-le;WANG Kun;FANG Xiang-ming(Anhui University of Chinese Medicine,Hefei 230012,China;the Second Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China;Anhui Province Key Laboratory of Application and Transformation of Traditional Chinese Medicine in Prevention and Treatment of Major Pulmonary Diseases,Hefei 230031,China;Key Laboratory of Xin'an Medicine,Ministry of Education,Hefei 230038,China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]安徽中医药大学第二附属医院,安徽合肥230012 [3]中医药防治肺系重大疾病应用转化安徽省重点实验室,安徽合肥230031 [4]新安医学教育部重点实验室,安徽合肥230038

出　　处：《中国中药杂志》2024年第24期6713-6723,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82074401);国家自然科学基金青年科学基金项目(82305187);安徽省教育厅自然科学研究重点项目(2022AH050446,2023AH050821);安徽中医药大学高层次人才支持计划项目(2024rcyb021);中医药防治肺系重大疾病应用转化安徽省重点实验室开放课题(2024ZYFBAHKLA09)。

摘　　要：探讨射干麻黄汤及其苦味药组通过肺-肠轴调控苦味受体14(TAS2R14)/分泌型免疫球蛋白A(SIgA)/胸腺基质淋巴细胞生成素(TSLP)干预寒哮大鼠上皮细胞屏障的机制。将50只SD大鼠随机分为5组,空白组、模型组、地塞米松组、射干麻黄汤组、苦味药组,10%鸡卵清白蛋白(OVA)腹腔及腹股沟两侧皮下注射致敏,联合2%OVA雾化及寒冷(2~4℃)刺激,激发大鼠寒哮模型。射干麻黄汤组、苦味药组、地塞米松组灌胃及雾化,空白组、模型组以生理盐水灌胃及雾化。末次激发后,对大鼠肺、肠组织进行苏木素-伊红(HE)染色、碘酸雪夫(PAS)染色观察肺、肠病理性变化;观察肺功能第1秒用力呼气量占用力肺活量比(FEV1/FVC)、用力呼气量为25%~75%肺活量时的平均流量/用力肺活量(FEV25%~75%/FVC)、最大呼气流量(PEF)及肺阻力(RL);酶联免疫吸附测定法(ELISA)检测血清中白细胞介素(IL)-4、IL-5、IL-13、肿瘤坏死因子(TNF)-α的含量,血清、肠与支气管黏膜中SIgA的含量;蛋白免疫印迹法检测大鼠肺组织TAS2R14蛋白表达情况;气相色谱-质谱联用法(GC-MS)测定大鼠粪便中短链脂肪酸(SCFAs)的含量;实验观察苦味药组含药血清通过TAS2R14/TSLP对脂多糖(LPS)诱导的上皮细胞炎症反应,蛋白免疫印迹法检测细胞中TAS2R14、TSLP蛋白表达。结果显示,与空白组相比,模型组大鼠饮水、饮食及体质量等均下降,肺与肠道炎性细胞浸润,杯状细胞增生,FEV1/FVC、FEV25%~75%/FVC、PEF均显著下降,RL显著上升,血清中IL-4、IL-5、IL-13和TNF-α含量均升高,血清、肠与支气管黏膜中SIgA均显著降低,肺组织中TAS2R14的表达抑制,粪便中乙酸、丙酸、丁酸含量均显著降低;LPS组TSLP表达上升,TAS2R14表达抑制。与模型组相比,射干麻黄汤、苦味药组大鼠一般情况均好转,肺与肠道炎性细胞浸润、杯状细胞增生均有所改善;FEV1/FVC、FEV25%~75%/FVC、PEF显著上升,RL显著下降;血清中LThis study aimed to investigate the mechanism by which Shegan Mahuang Decoction(SGMH) and its bitter Chinese herbs(BCHs) regulated the lung-gut axis through the bitter taste receptor 14(TAS2R14)/secretory immunoglobulin A(SIgA)/thymic stromal lymphopoietin(TSLP) to intervene in the epithelial cell barrier of cold asthma rats. Fifty SD rats were randomly divided into the following five groups: normal group, model group, dexamethasone group, SGMH group, and BCHs group. A 10% ovalbumin(OVA) solution was used to sensitize the rats via subcutaneous injection on both sides of the abdomen and groin, combined with 2% OVA atomization and cold(2-4 ℃) stimulation to induce a cold asthma model in rats. The SGMH, BCHs, and dexamethasone groups were given corresponding treatments by gavage and nebulization, while the normal and model groups received normal saline by gavage and nebulization. After the final stimulation, pathological changes in the lung and intestine tissues were observed using hematoxylin-eosin(HE) and periodic acid-Schiff(PAS) staining. Lung function was assessed by measuring the ratio of forced expiratory volume in the first second to forced vital capacity(FEV1/FVC), the ratio of the average flow rate at 25%-75% of forced vital capacity to foned vital capacity(FEV25%-75%/FVC), the peak expiratory flow(PEF), and pulmonary resistance(RL). The levels of IL-4, IL-5, IL-13, and TNF-α in serum, and sIgA in serum, intestinal, and bronchial mucosa were detected by enzyme-linked immunosorbent assay(ELISA). The expression of TAS2R14 protein in lung tissue was detected by Western blot(WB). The content of short-chain fatty acids(SCFAs) in rat feces was determined by gas chromatography-mass spectrometry(GC-MS). The effect of TAS2R14/TSLP on lipopolysaccharide(LPS)-induced inflammation in epithelial cells in the BCHs group was observed, and the expression of TAS2R14 and TSLP in cells was detected by WB. Compared with the normal group, the model group showed reduced water intake, diet, and body weight, increased infiltra

关 键 词：射干麻黄汤 苦味药 TAS2R14 SIGA SCFAs 

分 类 号：R28[医药卫生—中药学]