基于网络药理学和细胞实验研究硒代三角叶薯蓣次碱植生素减轻糖尿病肾病足细胞损伤的机制
Integrated Network Pharmacology and Cellular Assay to Explore the Mechanisms of Selenized Tripterine Phytosomes (Se@Tri-PTs) Alleviating Podocyte Injury in Diabetic Nephropathy.
机构信息
Department of Chinese Traditional Medicine, The First Affiliated Hospital of Jinan University, Guangzhou, 510630, People's Republic of China.
Department of Chinese Traditional Medicine, the First Affiliated Hospital of Jinan University, Guangzhou, 510630, People's Republic of China.
出版信息
Curr Pharm Des. 2023;29(38):3073-3086. doi: 10.2174/0113816128275079231102071508.
AIM
This work aimed to elucidate the mechanisms of Se@Tri-PTs in alleviating podocyte injury via network pharmacology and cellular assay.
BACKGROUND
Selenized tripterine phytosomes (Se@Tri-PTs) have been confirmed to undertake synergistic and sensitized effects on inflammation, which may be curatively promising for diabetic nephropathy (DN). However, the mechanisms of Se@Tri-PTs in alleviating podocyte injury, a major contributor to DN, still remain unclear.
OBJECTIVE
The objective of the study was to find out the underlying mechanisms of Se@Tri-PTs in alleviating podocyte injury in diabetic nephropathy.
METHODS
The key components and targets of significant for DN as well as the signaling pathways involved have been identified. A high glucose-induced podocyte injury model was established and verified by western blot. The protective concentration of Se@Tri-PTs was screened by CCK-8 assay. Podocytes cultured with high glucose were treated with Se@Tri-PTs under protective levels. The expression of key protective proteins, nephrin and desmin, in podocytes, was assayed by western blot. Further, autophagy- related proteins and factors, like NLRP3, Beclin-1, LC3II/LC3, P62, and SIRT1, were analyzed, which was followed by apoptosis detection.
RESULTS
Network pharmacology revealed that several monomeric components of , especially Tri, act on DN through multiple targets and pathways, including the NLRP3-mediated inflammatory pathway. Se@Tri-PTs improved the viability of podocytes and alleviated their injury induced by high glucose at 5 μg/L or above. High-glucose induction promoted the expression of NLRP3 in podocytes, while a low concentration of Se@Tri-PTs suppressed the expression. A long-term exposure of high glucose significantly inhibited the autophagic activity of podocytes, as manifested by decreased Beclin-1 level, lower ratio of LC3 II/LC3 I, and up- regulation of P62. This abnormality was efficiently reversed by Se@Tri-PTs. Importantly, the expression of SIRT1 was up-regulated and podocyte apoptosis was reduced.
CONCLUSION
Se@Tri-PTs can alleviate podocyte injury associated with DN by modulating NLRP3 expression through the pathway of SIRT1-mediated autophagy.
目的
本研究旨在通过网络药理学和细胞实验阐明硒化三尖杉酯碱植酸钠（Se@Tri-PTs）缓解足细胞损伤的作用机制。
背景
已证实硒化三尖杉酯碱植酸钠（Se@Tri-PTs）对炎症具有协同和敏化作用，这可能对糖尿病肾病（DN）具有治疗前景。然而，Se@Tri-PTs 缓解足细胞损伤的机制，即导致 DN 的主要因素，仍不清楚。
目的
本研究旨在探讨 Se@Tri-PTs 缓解糖尿病肾病足细胞损伤的作用机制。
方法
鉴定了与 DN 相关的关键成分和靶点，以及涉及的信号通路。通过 Western blot 验证了高糖诱导的足细胞损伤模型。通过 CCK-8 测定筛选 Se@Tri-PTs 的保护浓度。在保护性浓度下，用 Se@Tri-PTs 处理高糖培养的足细胞。通过 Western blot 检测足细胞中关键保护蛋白nephrin 和 desmin 的表达。进一步分析自噬相关蛋白和因子，如 NLRP3、Beclin-1、LC3II/LC3、P62 和 SIRT1，并进行凋亡检测。
结果
网络药理学显示， 中的几种单体成分，特别是三尖杉酯碱，通过多个靶点和通路作用于 DN，包括 NLRP3 介导的炎症通路。Se@Tri-PTs 可提高高糖诱导的足细胞活力，减轻其损伤，浓度为 5 μg/L 或以上。高糖诱导可促进足细胞中 NLRP3 的表达，而低浓度的 Se@Tri-PTs 可抑制其表达。长期高糖暴露可显著抑制足细胞的自噬活性，表现为 Beclin-1 水平降低、LC3 II/LC3 I 比值降低和 P62 上调，这些异常可被 Se@Tri-PTs 有效逆转。重要的是，SIRT1 表达上调，足细胞凋亡减少。
结论
Se@Tri-PTs 通过 SIRT1 介导的自噬途径调节 NLRP3 表达，缓解与 DN 相关的足细胞损伤。
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