背景:白芍(RPA),中药,经常用于治疗哮喘。先前的研究表明,与RPA(DCM)的二氯甲烷部分相比,搅拌油炸RPA(FDCM)的二氯甲烷部分增强了抗过敏性哮喘的作用。
目的:芍药苷(PF)的显着增加,没食子酸乙酯(EG),在FDCM中已观察到1,2,3,4,6-五酰葡萄糖(PGG)。本研究旨在探讨FDCM化合物在卵清蛋白(OVA)诱导的过敏性哮喘小鼠模型中的作用及机制。
方法:采用中压液相色谱(MPLC)富集FDCM中含量差异显著的化合物组分(FB-40)和其他组分。在OVA诱导的过敏性哮喘小鼠的所有部分中验证了药效学。此外,FB-40的药物剂量依赖性(0.42mg/kg,0.21mg/kg,和0.07mg/kg),这是FDCM中抗过敏哮喘最活跃的部分,被探索了。通过Western印迹分析分析IL-6、p-STAT3和STAT3的表达。此外,FB-40的主要成分通过UPLC标准进行鉴定。最后,通过LPS刺激的BEAS-2B细胞使用Elisa测定法检测FB-40主要成分的抗炎作用.
结果:结果显示FB-40是FDCM中最活跃的部分,能显著改善肺组织病理状况,并减少支气管肺泡灌洗液(BALF)中炎性细胞的数量。其具有比其主要成分PF更大的药理活性。FB-40在过敏性哮喘小鼠中也表现出剂量依赖性并调节IL-6/STAT3信号通路。此外,PF,Albiflorin(AF),PGG,EG,来自FB-40的1,2,3,6-四-O-没食子酰-β-D-葡萄糖(TGG)通过UPLC用标准品鉴定。最后,在LPS诱导的BEAS-2B细胞实验中,EG,PGG,1,2,3,6-Tetra-O-没食子酰-β-D-葡萄糖(TGG)显示出比单萜苷(PF和AF)更强的细胞因子抑制活性。
结论:研究证明FB-40是FDCM中的活性组分,调节IL-6/STAT3信号通路改善过敏性哮喘。没食子酸包括TGG和PGG,和EG也在治疗FB-40过敏性哮喘中发挥作用。
BACKGROUND: Radix Paeoniae Alba (RPA), a traditional Chinese medicine, has been used frequently in the treatment of asthma. Previous studies demonstrated the dichloromethane fraction of Stir-Frying RPA (FDCM) enhanced the effect of anti-allergic asthma compared with the dichloromethane fraction of RPA (DCM).
OBJECTIVE: The significant increasing of Paeoniflorin (PF), ethyl gallate (EG), 1,2,3,4,6-pentagalloylglucose (PGG) had been observed in FDCM. This study aimed to investigate the effects and mechanisms of these compounds from FDCM in ovalbumin (OVA)-induced allergic asthma mouse model.
METHODS: The significant difference contents compounds fraction (FB-40) and other fractions in FDCM were enriched by Medium Pressure Liquid Chromatography (MPLC). The pharmacodynamics was verified among all fractions in OVA-induced allergic asthma mice. Moreover, the drug dose dependence of FB-40 (0.42 mg/kg, 0.21 mg/kg, and 0.07 mg/kg), which were the most active fraction from FDCM for anti-allergic asthma, was explored. The expression of IL-6, p-STAT3, and STAT3 was analyzed by Western blot analysis. In addition, the main components of FB-40 were identified by UPLC with standards. Finally, the anti-inflammatory effects of the main components from FB-40 were detected by LPS-stimulated BEAS-2B cells using an Elisa assay.
RESULTS: The results showed that FB-40 was the most active fraction from FDCM, which could significantly improve the lung tissue pathological condition, and decrease the number of inflammatory cells in bronchoalveolar lavage fluid (BALF). It had greater pharmacological activity than its main component PF. FB-40 also showed dose dependence and regulated the IL-6/STAT3 signaling pathway in allergic asthma mice. Besides, PF, Albiflorin (AF), PGG, EG, and 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG) from FB-40 were identified by UPLC with the standard. At last, in the LPS-induced BEAS-2B cell experiments, EG, PGG, 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG) showed stronger inhibiting activities of cytokine than the monoterpenoid glycosides (PF and AF).
CONCLUSIONS: The research proved that FB-40 was an active fraction in FDCM, which regulates IL-6/STAT3 Signaling Pathway to ameliorate allergic asthma. Gallic acids including TGG and PGG, and EG also play a role in the treatment of allergic asthma in FB-40.