Fang Jing-Xian, Zhang Lian, Li Jing, Zhang Han-Rui, Liu Dan, Nie Jing, Ye Xiao-Chuan
Hubei Key Laboratory of Resource Science and Chemistry in Chinese Medicine,Pharmacy Faculty,Hubei University of Chinese Medicine Wuhan 430065,China.
Hubei Center for ADR Monitoring Wuhan 430071,China.
Zhongguo Zhong Yao Za Zhi. 2023 Aug;48(16):4446-4458. doi: 10.19540/j.cnki.cjcmm.20230320.401.
The present study aimed to explore the therapeutic effect and mechanism of non-polysaccharide fraction of Bletillae Rhizoma in the treatment of gastric ulcer by network pharmacology and animal experiments. UPLC-Q-TOF-MS/MS was employed to chara-cterize the chemical components of non-polysaccharide fraction of Bletillae Rhizoma, and the common targets of Bletillae Rhizoma and gastric ulcer were screened out by network pharmacology. The "drug-component-target-disease" network was constructed. Protein-protein interaction(PPI) network was established by STRING. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were performed based on Matescape database to predict the therapeutic effect and mechanism of Bletillae Rhizoma. Finally, the gastric ulcer model was induced in mice by alcohol to verify the therapeutic effect and mechanism of non-polysaccharide fraction of Bletillae Rhizoma on gastric ulcer. Forty-seven chemical components were identified from non-polysaccharide fraction of Bletillae Rhizoma, among which gymnoside Ⅰ, gymnoside Ⅱ, militarine, bletilloside A, and shancigusin I might be the main active components of non-polysaccharide fraction of Bletillae Rhizoma against gastric ulcer. PPI network analysis revealed core targets such as albumin(ALB), serine/threonine kinase 1(AKT1), tumor necrosis factor(TNF), and epidermal growth factor receptor(EGFR). The KEGG enrichment analysis showed that non-polysaccharide fraction of Bletillae Rhizoma mainly exerted the therapeutic effect by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway, mitogen-activated protein kinase(MAPK) signaling pathway, and Ras signaling pathway. The results of animal experiments showed that non-polysaccharide fraction of Bletillae Rhizoma could significantly improve alcohol-induced ulceration in mice to increase ulcer inhibition rate, decrease the levels of TNF-α, interleukin(IL)-1β, IL-6, vasoactive intestinal peptide(VIP), and thromboxane B2(TXB2), elevated the le-vels of IL-10, prostaglandin E2(PGE2), epidermal growth factor(EGF), and vascular endothelial growth factor(VEGF), down-re-gulate the protein levels of PI3K and AKT, and up-regulate the protein levels of p-PI3K and p-AKT. This study indicates that Bletillae Rhizoma may play a role in the treatment of gastric ulcer through multiple components, targets, and pathways and verifies partial prediction results of network pharmacology. The findings of this study provide a scientific and experimental basis for clinical application.
本研究旨在通过网络药理学和动物实验探讨白及非多糖部位治疗胃溃疡的作用效果及机制。采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）对白及非多糖部位的化学成分进行表征，并通过网络药理学筛选出白及与胃溃疡的共同靶点，构建“药物-成分-靶点-疾病”网络。利用STRING建立蛋白质-蛋白质相互作用（PPI）网络，基于Matescape数据库进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析，以预测白及的治疗作用及机制。最后，通过酒精诱导小鼠建立胃溃疡模型，验证白及非多糖部位对胃溃疡的治疗作用及机制。从白及非多糖部位鉴定出47种化学成分，其中裸茎苷Ⅰ、裸茎苷Ⅱ、白及胺、白及苷A和山慈菇苷Ⅰ可能是白及非多糖部位抗胃溃疡的主要活性成分。PPI网络分析揭示了白蛋白（ALB）、丝氨酸/苏氨酸激酶1（AKT1）、肿瘤坏死因子（TNF）和表皮生长因子受体（EGFR）等核心靶点。KEGG富集分析表明，白及非多糖部位主要通过调节磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号通路、丝裂原活化蛋白激酶（MAPK）信号通路和Ras信号通路发挥治疗作用。动物实验结果表明，白及非多糖部位可显著改善酒精诱导的小鼠溃疡，提高溃疡抑制率，降低TNF-α、白细胞介素（IL）-1β、IL-6、血管活性肠肽（VIP）和血栓素B2（TXB2）水平，升高IL-10、前列腺素E2（PGE2）、表皮生长因子（EGF）和血管内皮生长因子（VEGF）水平，下调PI3K和AKT蛋白水平，上调p-PI3K和p-AKT蛋白水平。本研究表明，白及可能通过多种成分、靶点和途径发挥治疗胃溃疡的作用，并验证了网络药理学的部分预测结果。本研究结果为临床应用提供了科学的实验依据。
