麻黄细辛附子汤对尘螨变应原Der p1诱导的鼻黏膜上皮细胞屏障损伤的影响  

Effects of Mahuang Xixin Fuzi Decoction on Dust Mite Allergen Der p1-induced Barrier Damage in Nasal Epithelial Cells

作　　者：倪钰莹 范淑月 崔颖[1] 高婧 罗再奕 刘敏[1] NI Yuying;FAN Shuyue;CUI Ying;GAO Jing;LUO Zaiyi;LIU Min(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《中国中医药信息杂志》2023年第10期101-107,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81774375)。

摘　　要：目的基于RhoA/ROCK信号通路探讨麻黄细辛附子汤及麻黄对尘螨变应原Der p1诱导的人鼻黏膜上皮细胞(HNEpC)屏障损伤的影响及作用机制。方法体外培养HNEpC,将细胞分为对照组、模型组、麻黄组、抑制剂组及麻黄细辛附子汤低、中、高剂量组。电阻仪检测细胞跨膜电阻,酶标仪检测FITCDextran通透性,倒置显微镜观察细胞形态,Western blot检测紧密连接蛋白咬合蛋白(Occludin)、闭合蛋白1(Claudin-1)及通路蛋白RhoA、ROCK1表达,免疫荧光染色检测肌动蛋白(F-actin)表达。结果与对照组比较,模型组细胞间隙增大,出现环形损伤,标准化跨膜电阻显著降低(P<0.01),FITC-Dextran通透性显著增加(P<0.01),Occludin、Claudin-1蛋白表达显著降低(P<0.01),RhoA、ROCK1、F-actin蛋白表达显著升高(P<0.01);与模型组比较,麻黄组、抑制剂组和麻黄细辛附子汤低、中、高剂量组细胞排列紧密,环形损伤被修复,标准化跨膜电阻显著升高(P<0.01),FITC-Dextran通透性显著降低(P<0.01),Occludin、Claudin-1蛋白表达显著升高(P<0.01),RhoA、ROCK1蛋白表达显著降低(P<0.01),麻黄细辛附子汤低、中、高剂量组和抑制剂组细胞F-actin蛋白表达显著降低(P<0.05,P<0.01)。结论麻黄细辛附子汤及麻黄可能通过抑制RhoA/ROCK信号通路改善鼻黏膜通透性,修复Der p1诱导的HNEpC屏障损伤。Objective To explore the effects and mechanism of Mahuang Xixin Fuzi Decoction and Ephedrae Herba on dust mite allergen Der p1-induced barrier damage in human nasal epithelial cells(HNEpC)based on RhoA/ROCK signaling pathway.Methods HNEpC were cultured in vitro.The cells were divided into control group,model group,Ephedrae Herba group,inhibitor group,and Mahuang Xixin Fuzi Decoction low-,medium-,high-dosage groups.Transepithelial resistance(TER)was measured by using the resistance measuring,FITC-Dextran permeability was detected by enzyme-labeled instrument,cell morphological changes were observed by inverted microscopy,Western blot was used to detect the expression levels of tight junction protein Occludin,Claudin-1,pathway protein RhoA and ROCK1,immunofluorescence staining was used to detect F-actin expression.Results Compared with the control group,the model group showed an increased cell gap,ring damage,significantly lower TER(P<0.01),significantly increased FITC-Dextran permeability(P<0.01),significantly lower Occludin and Claudin-1 protein expressions(P<0.01),and significantly higher RhoA,ROCK1,and F-actin protein expressions(P<0.01).Compared with the model group,cells were tightly arranged in Ephedrae Herba group,inhibitor group,and Mahuang Xixin Fuzi Decoction low-,medium-,high-dosage groups,ring damage was repaired,TER was significantly higher(P<0.01),FITCDextran permeability was significantly lower(P<0.01),Occludin and Claudin-1 protein expressions were significantly higher(P<0.01),RhoA and ROCK1 protein expressions significantly decreased(P<0.01),F-actin protein expression significantly decreased in Mahuang Xixin Fuzi Decoction low-,medium-,high-dosage groups and the inhibitor group(P<0.05,P<0.01).Conclusion Mahuang Xixin Fuzi Decoction and Ephedrae Herba may improve nasal mucosal permeability and repair Der p1-induced HNEpC barrier damage by inhibiting the RhoA/ROCK signaling pathway.

关 键 词：变应性鼻炎 麻黄细辛附子汤 屏障功能 RhoA/ROCK信号通路 人鼻黏膜上皮细胞 

分 类 号：R285.5[医药卫生—中药学]