白芍麸炒调控肥大细胞Gab2/AKT/Erk信号通路和宿主代谢标志物增强抗过敏性哮喘的物质基础和机制研究

Bran fryed Radix Paeoniae Alba have helps calming the liver and spleen tonic effect. It has significant curative effects to applying the Chinese medicine theory of Radix Paeoniae Alba to cure immune system diseases. Previous study discovered enhance the inhibitory effect on allergic asthma rat after processed. and until now there is no research on the mechanism of Chinese herbal pieces effect changes. In our preliminary experiment, Radix Paeoniae Alba after bran fried decrease of the expression of P-Gab2, P-Akt and P-Erk, and inhibit the mast cell degranulation and calcium release. Based on the previous researches, this subject advance hypothesis that “Radix Paeoniae Alba after bran fried through the regulation of Gab2/AKT/Erk signaling pathway to inhibit vesicles transport, mast cell degranulation and calcium release, thereby enhancing its anti-allergic asthma synergistic effect”. This subject through discuss the regulatory effect of overall metabolite and the regulation of key targets, using gene knockout, spectrum-effect relationship, metabonomics and other technology innovatively to study the correlation between the main constituents in vivo and the multitargeted of metabolic. By exploring the regulation of Gab2/AKT/Erk signaling pathway to clear the anti-allergic asthma active substance and reveal the multi-target synergistic mechanism of bran fried Radix Paeoniae Alba. This project provides the basis for the clinical application of bran fried Radix Paeoniae Alba and new thought for the traditional Chinese medicine processing theory research.

白芍具有养血柔肝、健脾益气功效，临床上常用于治疗免疫系统疾病。申请人前期研究发现，白芍具有抗过敏性哮喘作用，且麸炒后疗效增强，但其作用的物质基础和机理尚未明确。申请人研究发现，白芍麸炒后化学组分发生变化，且能够降低P-Gab2，P-Akt和P-Erk蛋白表达水平，抑制肥大细胞脱颗粒和钙流释放。因此提出科学假说“白芍麸炒后体内外化学组分发了转化，进而调控Gab2/AKT/Erk信号通路关键因子影响囊泡转运，抑制钙流和肥大细胞脱颗粒，发挥抑制过敏性哮喘增效作用”。本项目运用基因修饰、谱效关系和代谢组学等技术，通过整体辨识和靶标分析，结合体内物质基础，揭示白芍麸炒前后体内成分与代谢经路靶点的相关性，并探究其对Gab2/AKT/Erk信号关键因子调控作用，阐明白芍麸炒增强抗过敏性哮喘的物质基础和多靶点协同作用机制。本研究不仅有助于阐明白芍麸炒增效机制，同时也为白芍抗过敏性哮喘药物开发提供参考依据。

白芍具有养血柔肝、健脾益气的功效，临床上常用于治疗免疫系统疾病。申请人前期研究发现，白芍具有抗过敏性哮喘作用，且麸炒后疗效增强，但其物质基础和机理尚未明确。本研究发现白芍抗过敏性哮喘最强的活性部位是二氯甲烷部位。并且在蜜麸炒炮制过后，炮制品相同部位能通过调控Gab2/PI3K/AKT信号通路发挥增强抗过敏性哮喘的作用。经UPLC/Q-TOF-MS检测，炮制前后主要成分的含量具有显著性变化的化合物是PF、AF和PGG等。本课题发现这些显著性变化成分的保留时间主要集中在3-7 min，因此通过中压制备液相，将白芍麸炒二氯甲烷部位拆分为含量显著增加的成分段和其他成分段。后经药效学的验证，证明FB-40是药理作用最强的活性段，且呈现出药物剂量依赖性，且其作用效果不止依靠其主要化学成分PF，还与其他化合物有关。并且在对FB-40及其主要化合物通过LPS诱导的BEAS-2B支气管上皮损伤细胞模型实验中，发现TGG、PGG等鞣酸类化合物具有很强的抗支气管上皮损伤细胞损伤的活性。上述的研究阐明白芍中PF和PGG等活性成分通过调控Gab2/PI3K/AKT信号通路抑制炎症的发展进程，从而发挥抗过敏性哮喘的功效。本研究为白芍在过敏性哮喘的临床应用上奠定了一定的理论基础。