麻黄-杏仁药对对磷酸组胺诱导的RTE细胞损伤的保护作用

Protective Effect of Ephedra-Almond on RTE Cell Injury Induced by Histamine Phosphate

目的:观察麻黄-杏仁药对对大鼠气道细胞损伤的保护作用,探讨其缓解哮喘症状与调控EGFR、PI3K和Bax表达以及大分子通透性的相关作用。方法:选用U6(64)均匀设计表安排药对配比,以磷酸组胺孵育建立气道上皮细胞损伤模型,给予不同配比麻黄-杏仁药对的含药肠吸收液进行干预,采用RT-PCR检测气道上皮细胞EGFR、PI3K、Bax mRNA的表达,采用Western blotting检测气道上皮细胞EGFR、PI3K和Bax蛋白的表达,采用电阻测量仪检测跨细胞单层电阻值(TEER),采用荧光酶标仪检测FITC-BSA的荧光强度,采用荧光显微镜检测气道上皮细胞损伤模型对FITC-BSA的通透性。结果:麻黄-杏仁药对各组含药肠吸收液中,N1A-N5A均可下调EGFR和PI3K mRNA表达,N_(2)A和N_(3)A均可下调Bax mRNA表达;N_(1)A-N_(6)A均可下调EGFR蛋白相对含量,N_(3)A-N_(6)A均可下调PI3K蛋白含量,N_(1)A-N_(6)A均可下调Bax蛋白含量;N_(3)A、N_(5)A和N_(6)A均可增加TEER,N_(2)A-N_(6)A均可降低FITC-BSA的透过率。结论:麻黄-杏仁药对可下调EGFR、PI3K和Bax的表达,提高细胞膜完整性,增加TEER,降低大分子通透性,从而维持机体内环境稳态,抑制气道损伤,达到缓解哮喘症状的目的;进一步表明含药肠吸收液用于中药复方体外药理研究是客观、可行的。

Objective:To observe the protective effect of ephedra-almond on airway cell injury in rats,and to explore the relationship between asthma treatment and regulation of EGFR,PI3 K,Bax expression and macromolecular permeability.Methods:The injury model of airway epithelial cells was established by using U6(64)uniform design table,and the injury model of airway epithelial cells was established by histamine phosphate.The intestinal absorption solution containing different ratios of ephedra-almond was used to intervene.The expression of EGFR,PI3 Kand Bax mRNA in airway epithelial cells was detected by RT-PCR,the expression of EGFR,PI3 Kand Bax protein in airway epithelial cells was detected by Western blotting,and TEER,was detected by resistance meter.The fluorescence intensity of FITC-BSA was detected by fluorescence enzyme labeling instrument,and the permeability of airway epithelial cell injury model to FITC-BSA was detected by fluorescence microscope.Results:In the intestinal absorption solution of ephedra-almond,N1 A-N5 Acould down-regulate EGFR and PI3 KmRNA expression;N2 Aand N3 Acould down-regulate the expression of Bax mRNA;N1 A-N6 Acould down-regulate the relative content of EGFR protein;N3 A-N6 Acould down-regulate PI3 Kprotein content,and N1 A-N6 Acan down-regulate Bax Protein content;N3 A,N5 Aand N6 Acould increase TEER,N2 A-N6 A could decrease the transmittance of FITC-BSA.Conclusion:Ephedra-almond can down-regulate the expression of EGFR,PI3 Kand Bax,improve the integrity of cell membrane,increase TEER,and decrease the permeability of macromolecules,so as to maintain the homeostasis of internal environment,inhibit airway injury and relieve asthma symptoms,which further shows that the pharmacological study of traditional Chinese medicine compound prescription in vitro is objective and feasible.