肥胖已成为影响糖尿病等各种慢性疾病出现的全球性问题,发育不良,心脏病,和癌症。在这项研究中,在Murrayapaniculata活性化合物的代谢物分布与使用网络药理学探索脂肪组织的靶向机制之间开发了一种整合方法,分子对接,分子动力学模拟,和体外测试。使用块嵌套循环(BNL)的天际线查询技术获得的网络药理学结果表明,组蛋白乙酰转移酶p300(EP300),过氧化物酶体增殖物激活受体γ(PPARG),过氧化物酶体增殖物激活受体γ-辅激活因子1-α(PPARGC1A)是治疗肥胖的潜在靶点.这三种蛋白质的富集分析显示它们与肥胖有关,产热,能量代谢,脂肪细胞因子,脂肪细胞分化,和葡萄糖稳态。M.paniculata叶子的代谢物分析揭示了16种活性化合物,根据药物相似度和ADME结果选择了其中10种进行分子对接。PPARG和EP300与10种活性化合物的分子对接结果显示,所有对接中的结合亲和力值为≤-5.0kcal/mol,表明有很强的约束力。使用分子动力学模拟检查了对接产生的蛋白质-配体复合物的稳定性,我们观察到PPARG与反式-3-吲哚丙烯酸的最佳平均均方根偏差(RMSD)为0.99µ,低于天然配体BRL(2.02µ)。此外,EP300和天然配体99E的RMSD为2.70。和最低的RMSD与配体(1R,9S)-5-[(E)-2-(4-氯苯基)乙烯基]-11-(5-嘧啶基羰基)-7,11-二氮杂三环[7.3.1.02,7]十三碳-2,4-二烯-6-酮为3.33µA。验证M.paniculata治疗肥胖症潜力的体外试验表明,用M.paniculata乙醇提取物处理的3T3-L1成熟脂肪细胞中PPARG和EP300基因表达显着降低,浓度为62.5μg/ml和15.625μg/ml,分别。这些结果表明,M.paniculata可以通过破坏脂肪细胞成熟和影响细胞内脂质代谢来潜在地治疗肥胖症。
Obesity has become a global issue that affects the emergence of various chronic diseases such as diabetes mellitus, dysplasia, heart disorders, and cancer. In this study, an integration method was developed between the metabolite profile of the active compound of Murraya paniculata and the exploration of the targeting mechanism of adipose tissue using network pharmacology, molecular docking, molecular dynamics simulation, and in vitro tests. Network pharmacology results obtained with the skyline query technique using a block-nested loop (BNL) showed that histone acetyltransferase p300 (EP300), peroxisome proliferator-activated receptor gamma (PPARG), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A) are potential targets for treating obesity. Enrichment analysis of these three proteins revealed their association with obesity, thermogenesis, energy metabolism, adipocytokines, fat cell differentiation, and glucose homeostasis. Metabolite profiling of M. paniculata leaves revealed sixteen active compounds, ten of which were selected for molecular docking based on drug-likeness and ADME results. Molecular docking results between PPARG and EP300 with the ten active compounds showed a binding affinity value of ≤ -5.0 kcal/mol in all dockings, indicating strong binding. The stability of the protein-ligand complex resulting from docking was examined using molecular dynamics simulations, and we observed the best average root mean square deviation (RMSD) of 0.99 Å for PPARG with trans-3-indoleacrylic acid, which was lower than with the native ligand BRL (2.02 Å). Furthermore, the RMSD was 2.70 Å for EP300 and the native ligand 99E, and the lowest RMSD with the ligand (1R,9S)-5-[(E)-2-(4-Chlorophenyl)vinyl]-11-(5-pyrimidinylcarbonyl)-7,11-diazatricyclo[7.3.1.02,7]trideca-2,4-dien-6-one was 3.33 Å. The in vitro tests to validate the potential of M. paniculata in treating obesity showed that there was a significant decrease in PPARG and EP300 gene expressions in 3T3-L1 mature adipocytes treated with M. paniculata ethanolic extract starting at concentrations 62.5 μg/ml and 15.625 μg/ml, respectively. These results indicate that M. paniculata can potentially treat obesity by disrupting adipocyte maturation and influencing intracellular lipid metabolism.