胃复春治疗糖尿病性胃轻瘫作用机制预测及药效物质基础初探

Screening Weifuchun Capsules for the treatment of Functional Dyspepsia based on blood components and pharmacophore modelsMaterial basis

关键词:  糖尿病性胃轻瘫  胃复春胶囊  动物实验  网络药理学  分子对接  物质基础

Key words:  diabetic gastroparesis  Weifuchun capsules  animal experiments  network pharmacology  molecular docking  material basis

摘要:

目的：利用网络药理学、动物实验及分子生物学实验进行验证与分子对接,初步阐明胃复春治疗糖尿病性胃轻瘫(diabetic gastroparesis,DGP)的潜在作用机制及物质基础。方法：首先通过网络药理学筛选胃复春的主要有效化学成分及其作用靶点,获取DGP相关的疾病靶点。将二者的交集靶点进行基因本体(Gene Ontology,GO)功能注释和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。其次利用糖尿病胃轻瘫动物模型,检测胃动力能力相关指标,评价胃复春的药效,采用蛋白免疫印迹法(Western blot)验证网络药理学预测的关键靶点及通路。基于拓扑网络分析潜在关键化合物,并进行分子对接验证。结果：基于文献检索获取胃复春胶囊入血成分77个,TCMSP检索及SwissTargetPrediction预测得入血成分靶点663个。与DGP交集靶点76个,经蛋白互作网络分析显示TNF、IL6、INS、IL1B、ALB、IL2、PTGS2等为核心作用靶点。GO富集结果显示涉及生物过程1503种,细胞组成43种,分子功能98种。KEGG富集结果显示涉及通路107条,其中糖尿病并发症AGR受体信号通路、钙离子信号通路、C型凝集素受体信号通路、流体剪切应力和动脉粥样硬化、血脂和动脉粥样硬化或为胃复春起效的核心通路。与模型组相比,给药组大鼠体质量均明显增加；与模型组相比,各给药组大鼠胃排空率、肠道推进率以及MTL、GAS、SP均显著增高(P<0.05)。挑选网络药理学预测的作用蛋白进行Western blot实验验证,发现胃复春干预组大鼠INS、MAPK、TNF、NOS3、IL1B相对表达水平均有显著改变。潜在活性物质基础蓝萼甲素、蓝萼乙素、Melissoidesin U、异鼠李素等与核心作用靶点对接良好。结论：胃复春可通过作用于INS、MAPK、TNF、NOS3、ILB等靶点,调节糖尿病并发症AGR受体等信号通路,发挥改善DGP模型大鼠胃肠动力、调节糖脂代谢紊乱、减少微血管损伤等作用,其潜在的药效物质基础可能为蓝萼甲素、蓝萼乙素、Melissoidesin U、异鼠李素等。

Abstract:

Objective: Use network pharmacology, animal experiments and molecular biology experiments to conduct verification and molecular docking to preliminarily elucidate the potential mechanism and material basis of Weifuchun in the treatment of diabetic gastroparesis (DGP). Methods: First, network pharmacology was used to screen the main effective chemical components and action targets of Weifuchun to obtain DGP-related disease targets. The intersection targets of the two were subjected to Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Secondly, an animal model of diabetic gastroparesis was used to detect indicators related to gastric motility capacity, evaluate the efficacy of gastric rejuvenation, and use Western blot to verify the key targets and pathways predicted by network pharmacology. Potential key compounds were analyzed based on topological networks and verified by molecular docking. Results: Based on the literature search, 77 blood components of Weifuchun Capsules were obtained, and 663 blood component targets were predicted by TCMSP search and SwissTargetPrediction. There are 76 targets that intersect with DGP, and protein interaction network analysis shows that TNF, IL6, INS, IL1B, ALB, IL2, PTGS2, etc. are the core targets. The GO enrichment results showed that 1503 types of biological processes, 43 types of cellular components, and 98 types of molecular functions were involved. KEGG enrichment results show that 107 pathways are involved, including diabetic complications AGR receptor signaling pathway, calcium ion signaling pathway, C-type lectin receptor signaling pathway, fluid shear stress and atherosclerosis, blood lipids and atherosclerosis It may be the core pathway for the effect of stomach rejuvenation. Compared with the model group, the body weight of rats in the administration group increased significantly; compared with the model group, the gastric emptying rate, intestinal propulsion rate, MTL, GAS, and SP of rats in each administration group were significantly increased (P< 0.05). Western blot experiments were performed on selected proteins predicted by network pharmacology, and it was found that the relative expression levels of INS, MAPK, TNF, NOS3, and IL1B in rats in the Weifuchun intervention group were significantly changed. The basic potential active substances include bluecalyxin A, bluecalyxine B, Melissoidesin U, isorhamnetin, etc., which are well connected with the core targets. Conclusion: Weifuxun can regulate diabetic complications AGR receptor and other signaling pathways by acting on INS, MAPK, TNF, NOS3, ILB and other targets, thereby improving gastrointestinal motility, regulating glucose and lipid metabolism disorders, and reducing diabetes complications in DGP model rats. Microvascular damage and other effects, its potential pharmacological material basis may be cyanacaldin A, cyanacaldin B, Melissoidesin U, isorhamnetin, etc.