背景:干姜-/葡萄酒-/柳叶黄连(CR,zCR/wCR/eCR)是临床上常用的CR加工产品。用不同的赋形剂加工后,CR的疗效会相应改变。I.e.,wCR可以解决上部激励器的过热,ZCR可以消除胃热,协调胃,eCR可以平滑肝脏和协调胃。然而,潜在机制尚不清楚.
目的:进一步验证3种CR产品的疗效差异,并比较其治疗胃溃疡的机制。
方法:首先,GU模型,其发病与胃热和肝胃不和密切相关,成立了,通过病理观察和细胞因子水平的测量来评价zCR/wCR/eCR/CR的治疗效果。第二,通过代谢组学分析和网络药理学揭示zCR/eCR对GU的差异干预机制。第三,代谢组学分析和网络药理学的预测机制使用蛋白质印迹进行了验证,流式细胞术和免疫荧光。
结果:zCR/wCR/eCR/CR可以不同程度地减轻病理损伤。在代谢组学研究中,血清样本中富集的代谢途径较少,其中大部分也存在于胃组织样本的结果中。胃保护剂,抗炎,抗氧化剂,zCR/wCR/eCR/CR的抗凋亡作用可能是由于它们对组氨酸的干扰,花生四烯酸,和甘油磷脂代谢。定量结果表明,zCR/eCR治疗GU的疗效优于wCR/CR。代谢组学和网络药理学的综合分析显示,zCR和eCR通过干预5个核心靶点发挥抗GU作用。包括AKT,TNF-α,IL6、IL1B和PPARG。在验证实验中,zCR/eCR可以显著逆转细胞凋亡相关蛋白的异常表达,炎症,氧化应激,胃功能,以及PI3K/AKT信号通路。
结论:zCR和eCR可以通过抵抗炎症和凋亡来提供胃保护作用,抑制胃酸分泌,以及加强胃粘膜防御和抗氧化能力。整合网络药理学和代谢组学分析可以揭示药物的作用机制,促进抗GU药物的发展。
BACKGROUND: Zingiberis rhizoma recens-/wine-/euodiae fructus-processed Coptidis Rhizoma (CR, zCR/wCR/eCR) are the commonly used processed products of CR in clinic. After being processed with different excipients, the efficacy of CR will change accordingly. I.e., wCR could resolve excessive heat of the upper energizer, zCR could eliminate gastric heat and harmonize the stomach, eCR could smooth the liver and harmonize the stomach. However, the underlying mechanisms were still unclear.
OBJECTIVE: To further verify the differential efficacy of the three processed CR products and compare the mechanisms on gastric ulcer.
METHODS: First, a GU model, whose onset is closely related to the heat in stomach and the disharmony between liver and stomach, was established, and the therapeutic effects of zCR/wCR/eCR/CR were evaluated by pathologic observation and measurement of cytokine levels. Second, metabolomics analysis and network pharmacology were conducted to reveal the differential intervening mechanism of zCR/eCR on GU. Third, the predicted mechanisms from metabolomics analysis and network pharmacology were validated using western blotting, flow cytometry and immunofluorescence.
RESULTS: zCR/wCR/eCR/CR could alleviate the pathologic damage to varying degrees. In metabolomics research, fewer metabolic pathways were enriched in serum samples, and most of them were also present in the results of gastric tissue samples. The gastroprotective, anti-inflammatory, antioxidant, and anti-apoptotic effects of zCR/wCR/eCR/CR might be due to their interference on histidine, arachidonic acid, and glycerophospholipids metabolism. Quantitative results indicated that zCR/eCR had a better therapeutic effect than wCR/CR in treating GU. A comprehensive analysis of metabolomics and network pharmacology revealed that zCR and eCR exerted anti-GU effects via intervening in five core targets, including AKT, TNF, IL6, IL1B and PPARG. In the validation experiment, zCR/eCR could significantly reverse the abnormal expression of proteins related to apoptosis, inflammation, oxidative stress, gastric function, as well as the PI3K/AKT signaling pathways.
CONCLUSIONS: zCR and eCR could offer gastroprotective benefits by resisting inflammation and apoptosis, inhibiting gastric-acid secretion, as well as strengthening gastric mucosal defense and antioxidant capacity. Integrating network pharmacology and metabolomics analysis could reveal the acting mechanism of drugs and promote the development of medications to counteract GU.