Withanolide A抑制hIAPP聚集：基于计算机、生物物理和果蝇的体内验证
Withanolide A inhibits hIAPP aggregation: An In silico, biophysical, and drosophila-based In vivo validation.
摘要(中文译文)
人胰岛淀粉样多肽（hIAPP）或胰淀素异常聚集形成毒性寡聚体和纤维，导致胰腺β细胞功能障碍及进行性细胞死亡，这是II型糖尿病（T2DM）的关键病理特征。本研究采用虚拟筛选与分子动力学（MD）模拟相结合的策略，并结合实验验证，以寻找hIAPP聚集抑制剂。我们通过分子对接从天然产物中筛选了2000种植物化学成分，随后进行计算机ADMET预测。发现Withaferin A和Withanolide A（南非醉茄的植物化学成分）是具有合适类药特性的先导分子。接下来，我们进行MD模拟以评估hIAPP-配体复合物的稳定性和相互作用动力学，以及配体对hIAPP纤维的影响。在计算筛选的基础上，我们进一步开展了全面的实验分析以验证先导分子的抑制作用。硫黄素T（ThT）测定、FTIR实验以及共聚焦和透射电子显微镜（TEM）的综合实验结果表明，配体（尤其是Withanolide A）通过延长滞后期、减少β-折叠含量和抑制hIAPP纤维形成，对hIAPP聚集具有强效抑制作用。在使用果蝇模型进行的体内验证中，通过多种细胞死亡染色测定和活性氧（ROS）分析证实，Withanolide A可通过减少果蝇肠道中的凋亡、坏死和氧化应激来减轻hIAPP寡聚体诱导的毒性。此外，在糖尿病果蝇中，Withanolide A降低了葡萄糖水平，表现出抗糖尿病活性。因此，本研究首次提出Withanolide A可能是抑制hIAPP聚集的潜在候选物及T2DM治疗药物。
Abstract
The aberrant aggregation of human islet amyloid polypeptide (hIAPP) or Amylin into toxic oligomers and fibrils leads to pancreatic β-cell dysfunction and progressive cell death, which is a key pathological feature of Type II diabetes mellitus (T2DM). In this study, we adopted a hybrid approach combining virtual screening and molecular dynamics (MD) simulation, with experimental validation, to identify inhibitors of hIAPP aggregation. Herein, we screened 2000 phytoconstituents from natural products using molecular docking, followed by in silico ADMET predictions. Withaferin A and Withanolide A (phytoconstituents of Ashwagandha) were found to be lead molecules with suitable drug-like properties. Next, we performed MD simulation to assess the stability and interaction dynamics of hIAPP-ligand complexes, and the effect of ligands on hIAPP fibrils. Building on the computational screening, we further carried out a comprehensive experimental analysis to validate the inhibitory effects of lead molecules. The collective experimental results from the Thioflavin T (ThT) assays, FTIR experiment, combined with Confocal and Transmission Electron Microscopy (TEM), suggest that the ligands (preferably Withanolide A) have a potent inhibitory effect against hIAPP aggregation by increasing the lag phase, reducing β-sheet content, and inhibiting fibril formation of hIAPP. For in vivo validation using Drosophila model, Withanolide A was found to mitigate hIAPP oligomer-induced toxicity by reducing apoptosis, necrosis, and oxidative stress in the Drosophila gut, as confirmed by multiple cell death staining assays and reactive oxygen species (ROS) analysis. Besides, in diabetic flies, Withanolide A lowered glucose levels, demonstrating anti-diabetic activity. Thus, this work, for the first time, suggests that Withanolide A may be a potential candidate for inhibiting hIAPP aggregation and as a T2DM drug.
如何引用
AMA
Smita Manjari Panda, Kalpanarani Dash, Devi Prashanna Behera, Monalisa Mishra, Harekrushna Sahoo, Umakanta Tripathy. Withanolide A inhibits hIAPP aggregation: An In silico, biophysical, and drosophila-based In vivo validation.. Scientific reports. 2026; doi:10.1038/s41598-026-60648-x.
APA
Smita Manjari Panda, Kalpanarani Dash, Devi Prashanna Behera, Monalisa Mishra, Harekrushna Sahoo, Umakanta Tripathy (2026). Withanolide A inhibits hIAPP aggregation: An In silico, biophysical, and drosophila-based In vivo validation.. Scientific reports. https://doi.org/10.1038/s41598-026-60648-x
GB/T 7714
Smita Manjari Panda, Kalpanarani Dash, Devi Prashanna Behera, Monalisa Mishra, Harekrushna Sahoo, Umakanta Tripathy. Withanolide A inhibits hIAPP aggregation: An In silico, biophysical, and drosophila-based In vivo validation.[J]. Scientific reports, 2026 doi:10.1038/s41598-026-60648-x.
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