Mechanistic insights of Cucumis melo L. seeds for gastrointestinal muscle spasms through calcium signaling pathway-related gene regulation networks in WGCNA and in vitro, in vivo studies
Mechanistic insights of Cucumis melo L. seeds for gastrointestinal muscle spasms through calcium signaling pathway-related gene regulation networks in WGCNA and in vitro, in vivo studies
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DOI：
10.1016/j.compbiomed.2023.106596
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发表时间：
2023-02-09
影响因子：
7.7
通讯作者：
Gandara，Jesus Simal
中科院分区：
文献类型：
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作者：
Wahid，Muqeet;Saqib，Fatima;Gandara，Jesus Simal
背景除了黄瓜的营养益处之外，巴基斯坦和印度的草药医生用种子来治疗各种疾病。本研究旨在确定C. meleseeds在钙介导的平滑肌收缩。采用LC ESI-MS/MS和HPLC技术测定其对平滑肌的舒张作用。然后，从NCBI GEO获得以大肠杆菌为主的肠易激综合征基因数据集，DEG和WGCNA随后进行功能富集分析。然后对关键基因进行分子对接。结果表明，苦瓜种子中芦丁、山萘酚和槲皮素的含量分别为702.38 μg/g、686.29 μg/g和658.41 μg/g。苦豆子对氯化钾（80 mM）诱导的痉挛性收缩具有剂量依赖性舒张作用，并在钙剂量-反应曲线中显示钙拮抗反应。在体内研究中，Cm.EtOH表现出抗腹泻、抗蠕动和抗分泌作用。WGCNA和DEG IBS相关致病基因的功能富集，包括参与钙介导的信号传导、MAPK级联和炎症反应的基因。通过分子对接，发现MAPK 1和PIK 3CG是与芦丁、槲皮苷和山奈酚结合能力较强的关键基因。通过改变钙介导的平滑收缩的调节基因产生的。
BackgroundIn addition to the nutritional benefits ofCucumis meloL., herbalists in Pakistan and India employ seeds to treat various ailments. This study aimed to determine the regulatory role ofC. meloseeds in calcium-mediated smooth muscle contraction.MethodsWe identified and quantified the phytochemicals ofC. melowith LC ESI–MS/MS and HPLC, then conductedin vitroandin vivotests to confirm the involvement in smooth muscle relaxation. Then, diarrhea-predominant irritable bowel syndrome gene datasets from NCBI GEO were acquired, DEGs and WGCNA followed by functional enrichment analysis. Next, molecular docking of key genes was performed.ResultsThe quantification ofC. meloseeds revealed concentrations of rutin, kaempferol, and quercetin were 702.38 μg/g, 686.29 μg/g, and 658.41 μg/g, respectively.In vitroexperiments revealed thatC. meloseeds had a dose-dependent relaxant effect for potassium chloride (80 mM)–induced spastic contraction and exhibited calcium antagonistic response in calcium dose-response curves. Inin vivostudies, Cm.EtOH exhibited antidiarrheal, antiperistaltic, and antisecretory effects. The functional enrichment of WGCNA and DEGs IBS-associated pathogenic genes, including those involved in calcium-mediated signaling, MAPK cascade, and inflammatory responses. MAPK1 and PIK3CG were identified as key genes with greater binding affinity with rutin, quercitrin, and kaempferol in molecular docking.ConclusionsThe bronchodilator and antidiarrheal effects ofC. melowere produced by altering the regulatory genes of calcium-mediated smooth contraction.