基于分子对接及分子动力学模拟揭示植物甾醇治疗慢性肾脏病的作用机制

doi:10.5246/jcps.2026.05.029

摘要/Abstract

摘要：

慢性肾脏病(CKD)因其高发病率、复杂的发病机制及有限的治疗手段，已成为全球亟待应对的重大公共健康问题。植物甾醇是一类广泛存在于植物中的天然活性化合物，因其抗炎、抗氧化及免疫调节作用而受到广泛关注。其中，豆甾醇和β-谷甾醇是最主要且生物活性较高的代表性成分，既往研究表明它们对心血管疾病、代谢性疾病及肾脏疾病具有潜在保护作用。本研究在叶酸诱导的CKD小鼠模型中，探讨了其可能的作用机制。转录组分析显示，先天免疫相关基因Sting1在CKD模型组中显著上调，而在植物甾醇干预后下调。GO和KEGG富集分析提示，其治疗作用可能涉及细胞因子-受体相互作用及趋化因子信号通路等免疫和炎症相关通路。分子对接及分子动力学模拟结果表明，两种植物甾醇均可稳定结合Sting1蛋白，其中豆甾醇表现出更强的结合能力和更优的构象稳定性。RMSD、RMSF及Rg分析结果表明复合物具有良好的构象稳定性，MM-PBSA能量分解进一步识别出PHE-268和PRO-208为关键的结合位点。此外，在HK-2细胞中进行的细胞热稳定性实验(CETSA)结果显示，两种化合物可在中度热应激(40–49 °C)下增强Sting1蛋白的热稳定性，进一步证实了其直接结合作用。综上，植物甾醇可能是通过靶向Sting1蛋白而发挥肾脏保护作用。本研究为植物甾醇在CKD治疗中的应用提供了新的作用机制见解和理论依据。

关键词: 慢性肾脏病, 植物甾醇, 分子对接, 分子动力学模拟, Sting1

Abstract:

Chronic kidney disease (CKD) represents a growing global health challenge due to its high prevalence, multifactorial pathogenesis, and limited therapeutic options. Phytosterols, a class of naturally occurring bioactive compounds widely present in plants, have garnered increasing attention for their anti-inflammatory, antioxidant, and immunomodulatory properties. Among these, stigmasterol (Stig) and β-sitosterol (β-Sito) are the most abundant and biologically active representatives, previously reported to confer protective effects against cardiovascular, metabolic, and renal disorders. In the present study, we explored the potential molecular mechanisms of these phytosterols in a folic acid (FA)-induced CKD mouse model. Comprehensive transcriptomic analysis revealed that Sting1, a gene critically involved in innate immune responses, was markedly upregulated in CKD and significantly downregulated following phytosterol treatment. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses further implicated immune- and inflammation-related pathways, such as cytokine-cytokine receptor interaction and chemokine signaling, as potential mediators of the therapeutic effects. Molecular docking and molecular dynamics (MD) simulations demonstrated stable binding of both phytosterols to the Sting1 protein, with stigmasterol exhibiting higher binding affinity and greater conformational stability. Detailed analyses of root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), and radius of gyration (Rg) supported the structural integrity of the phytosterol-Sting1 complexes. MM-PBSA energy decomposition identified PHE-268 and PRO-208 as key residues mediating the interactions. Moreover, cellular thermal shift assays (CETSA) in HK-2 cells confirmed that both compounds enhanced the thermal stability of Sting1 under moderate heat stress (40–49 °C), providing additional evidence for a direct interaction. Taken together, these findings indicated that phytosterols might exert nephroprotective effects through direct modulation of Sting1, thereby attenuating immune and inflammatory dysregulation in CKD. This study offered novel mechanistic insights and established a theoretical foundation for the development of phytosterol-based therapeutic interventions for CKD.

Key words: Chronic kidney disease, Phytosterols, Molecular docking, Molecular dynamics simulations, Sting1

Supporting:

李睿, 张影, 陈聿荣, 高映杰, 杨帆. 基于分子对接及分子动力学模拟揭示植物甾醇治疗慢性肾脏病的作用机制[J]. 中国药学(英文版), 2026, 35(5): 407-419.

Rui Li, Ying Zhang, Yurong Chen, Yingjie Gao, Fan Yang. Molecular docking and dynamics simulations reveal phytosterol mechanisms in chronic kidney disease therapy[J]. Journal of Chinese Pharmaceutical Sciences, 2026, 35(5): 407-419.

图/表 8

Figure 1. Volcano plots of gene expression changes in renal tissue samples from each group. (A) Normal group vs. Model group; (B) Model group vs. Stig-treated group; (C) Model group vs. β-Sito-treated group.

Figure 2. Venn diagram of DEGs and functional enrichment analysis. (A) Venn diagram of DEGs; (B−D) GO functional enrichment analysis of DEGs; (E) KEGG pathway enrichment analysis of DEGs.

Figure 3. Molecular docking results. (A) Results of Stig with Sting1; (B) Results of β-Sito with Sting1.

Figure 4. RMSD and RMSF plots of Stig-Sting1 and β-Sito-Sting1. (A) RMSD plot of Stig-Sting1; (B) RMSD plot of β-Sito-Sting1; (C) RMSF plot of Stig-Sting1; (D) RMSF plot of β-Sito-Sting1.

Figure 5. Rg plot of Stig-Sting1 and β-Sito-Sting1. (A) Rg plot of Stig-Sting1; (B) Rg plot of β-Sito-Sting1.

Table 1. MM-PBSA of Stig-Sting1 and β-Sito-Sting1.

Figure 6. Residue decomposition plot of the binding energy of Stig-Sting1 and β-Sito-Sting1. (A) Stig-Sting1; (B) β-Sito-Sting1.

Figure 7. CETSA results of HK-2 cells. (A) CETSA results of Stig; (B) CETSA results of β-Sito.

参考文献 18

[1]	Chen, T.K.; Knicely, D.H.; Grams, M.E. Chronic kidney disease diagnosis and management: a review. JAMA. 2019, 322, 1294–1304.
[2]	Borg, R.; Carlson, N.; Søndergaard, J.; Persson, F. The growing challenge of chronic kidney disease: an overview of current knowledge. Int. J. Nephrol. 2023, 2023, 9609266.
[3]	Shen, M.Y.; Yuan, L.L.; Zhang, J.; Wang, X.F.; Zhang, M.Y.; Li, H.Z.; Jing, Y.; Zeng, F.J.; Xie, J.H. Phytosterols: physiological functions and potential application. Foods. 2024, 13, 1754.
[4]	Suttiarporn, P.; Chumpolsri, W.; Mahatheeranont, S.; Luangkamin, S.; Teepsawang, S.; Leardkamolkarn, V. Structures of phytosterols and triterpenoids with potential anti-cancer activity in bran of black non-glutinous rice. Nutrients. 2015, 7, 1672–1687.
[5]	Ramprasath, V.R.; Awad, A.B. Role of phytosterols in cancer prevention and treatment. J. AOAC Int. 2015, 98, 735–738.
[6]	Shahzad, N.; Khan, W.; Shadab, M.D.; Ali, A.; Saluja, S.S.; Sharma, S.; Al-Allaf, F.A.; Abduljaleel, Z.; Abdel Aziz Ibrahim, I.; Abdel-Wahab, A.F.; Afify, M.A.; Al-Ghamdi, S.S. Phytosterols as a natural anticancer agent: Current status and future perspective. Biomed. Pharmacother. 2017, 88, 786–794.
[7]	Gumede, N.M.; Lembede, B.W.; Nkomozepi, P.; Brooksbank, R.L.; Erlwanger, K.H.; Chivandi, E. Protective effect of β-sitosterol against high-fructose diet-induced oxidative stress, and hepatorenal derangements in growing female sprague-dawley rats. Lab. Anim. Res. 2024, 40, 30.
[8]	Yang, F.; Sun, L.; Gao, Y.J.; Liang, J.Z.; Ye, W.Q.; Yang, W.J.; Xie, S.Y.; Zhou, J.T.; Li, R.S. Integrating network pharmacology and metabolomics to explore the potential mechanism of β-sitosterol against hyperuricemia nephropathy. J. Food Biochem. 2024, 2024, 7645677.
[9]	Yang, F.; Gao, Y.J.; Xie, S.Y.; Yang, W.J.; Wang, Q.Y.; Ye, W.Q.; Sun, L.; Zhou, J.T.; Feng, X.E. Dietary phytosterol supplementation mitigates renal fibrosis via activating mitophagy and modulating the gut microbiota. Food Funct. 2025, 16, 2316–2334.
[10]	Antwi, A.O.; Obiri, D.D.; Osafo, N.; Forkuo, A.D.; Essel, L.B. Stigmasterol inhibits lipopolysaccharide-induced innate immune responses in murine models. Int. Immunopharmacol. 2017, 53, 105–113.
[11]	Qian, K.; Zheng, X.X.; Wang, C.; Huang, W.G.; Liu, X.B.; Xu, S.D.; Liu, D.K.; Liu, M.Y.; Lin, C.S. β-sitosterol inhibits rheumatoid synovial angiogenesis through suppressing VEGF signaling pathway. Front. Pharmacol. 2022, 12, 816477.
[12]	Tang, X.L.; Yan, T.; Wang, S.Y.; Liu, Q.Q.; Yang, Q.; Zhang, Y.Q.; Li, Y.J.; Wu, Y.M.; Liu, S.B.; Ma, Y.L.; Yang, L. Treatment with β-sitosterol ameliorates the effects of cerebral ischemia/reperfusion injury by suppressing cholesterol overload, endoplasmic reticulum stress, and apoptosis. Neural Regen. Res. 2024, 19, 642–649.
[13]	Deng, L.; Guo, S.J.; Liu, Y.P.; Zhou, Y.J.; Liu, Y.R.; Zheng, X.X.; Yu, X.J.; Shuai, P. Global, regional, and national burden of chronic kidney disease and its underlying etiologies from 1990 to 2021: a systematic analysis for the Global Burden of Disease Study 2021. BMC Public Health. 2025, 25, 636.
[14]	Sang, Y.H.; Yun, J.; Dai, L.J.; Song, L.Q. Chinese herbal enema therapy for the treatment of chronic kidney disease. J. Chin. Pharm. Sci. 2024, 33, 305–315.
[15]	Francis, A.; Harhay, M.N.; Ong, A.C.M.; Tummalapalli, S.L.; Ortiz, A.; Fogo, A.B.; Fliser, D.; Roy-Chaudhury, P.; Fontana, M.; Nangaku, M.; Wanner, C.; Malik, C.; Hradsky, A.; Adu, D.; Bavanandan, S.; Cusumano, A.; Sola, L.; Ulasi, I.; Jha, V.; American Society of Nephrology, European Renal Association & International Society of Nephrology. Chronic kidney disease and the global public health agenda: an international consensus. Nat. Rev. Nephrol. 2024, 20, 473–485.
[16]	Brüll, F.; Mensink, R. Plant sterols: functional lipids in immune function and inflammation? Clin. Lipidol. 2009, 4, 355–365.
[17]	Zhang, R.X.; Kang, R.; Tang, D.L. The STING1 network regulates autophagy and cell death. Signal Transduct. Target. Ther. 2021, 6, 208.
[18]	Maekawa, H.; Fain, M.E.; Wasano, K. Pathophysiological roles of the cGAS-STING inflammatory pathway. Physiology. 2023, 38, 167–177.