基于非靶向代谢组学和肠道菌群探究经典名方泻白散对过敏性哮喘大鼠的保护作用

Protective effect of Xiebaisan on allergic asthma in rats based on non-targeted metabolomics and intestinal bacterial flora

目的本研究拟从宿主-肠道菌群-代谢的角度探讨泻白散保护过敏性哮喘大鼠可能的微观机制。方法将SPF级SD大鼠分为正常对照组(NC组)、过敏性哮喘模型组(M组)和泻白散组(Xiebaisan组)。通过卵清白蛋白(OVA)诱导建立大鼠过敏性哮喘模型;HE染色观察大鼠肺组织病理学变化;取结肠内容物进行16S rDNA高通量测序,观察肠道菌群结构与功能的变化;采用超高效液相色谱-四级杆飞行时间串联质谱法(UHPLC-Q-TOF/MS)分别进行血清样本和肺组织样本非靶向代谢组学检测。结果HE染色显示:与NC组相比,Xiebaisan组可以在一定程度上改善哮喘大鼠肺部组织形态结构;16S rDNA高通量测序结果显示:M组肠道菌群多样性降低,与M组相比,Xiebaisan组肠道菌群多样性增加,肠道微生态得以改善;血清非靶向代谢组学检测结果显示:Xiebaisan组能够调节氨基酸代谢、mTOR等通路,且部分回调M组引起的差异代谢物表达;肺组织样本非靶向代谢组学检测结果显示,Xiebaisan组能够调节碳代谢、血管平滑肌收缩信号通路和cAMP等信号通路,且部分回调M组引起的差异代谢物表达。结论泻白散可能通过改善肺组织形态结构、肠道菌群的多样性以及调控mTOR通路、血管平滑肌收缩信号通路和cAMP等信号通路影响氨基酸代谢、碳代谢等途径发挥对过敏性哮喘大鼠的保护作用。

Objective To explore the possible mechanism of Xiebaisan in protecting against allergic asthma in rats from the perspective of host intestinal flora metabolism.Methods SPF SD rats were divided into normal group(NC group),model group(M group),and Xiebaisan group.The allergic asthma rat model was established by ovalbumin.Changes in lung histopathology were observed by HE staining.Colon contents were harvested for 16S rDNA high-throughput sequencing to assess changes in the intestinal flora structure and function.Serum and lung tissue samples were collected for non-targeted metabolomics by Ultra-high performance liquid-time-of-flight mass spectrometer.Results HE staining showed some improvement of lung histomorphology in asthmatic rats in the Xiebaisan group compared with that in the M group.16S rDNA high-throughput sequencing showed that the diversity of intestinal flora was decreased in the M group and increased in the Xiebaisan group compared with the M group,the microecosystem of intestinal was improved.Non-targeted metabolomics of serum showed regulation of amino acid metabolism and the mTOR pathway in the Xiebaisan group,and partially reversed differential metabolite expression in the M group.Non-targeted metabonomics of lung tissue samples showed regulation of carbon metabolism,vascular smooth muscle and cAMP signaling pathways in the Xiebaisan group,and partially reversed differential metabolite expression in the M group.Conclusions The protective effects of Xiebaisan on allergic asthma in rats may be related to improvement of the morphological structure of lung tissue,the diversity of intestinal flora,and regulation of mTOR,vascular smooth muscle contraction,and cAMP pathways,which affect amino acid and carbon metabolism.