南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (3): 465-473.doi: 10.12122/j.issn.1673-4254.2024.03.07

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健脾滋肾方抑制系统性红斑狼疮患者的足细胞自噬：基于网络药理学和临床研究

陈君洁，黄传兵，李 明   

  1. 安徽中医药大学第一临床医学院，安徽 合肥 230000；安徽中医药大学第一附属医院风湿免疫科，安徽 合肥 230031
  • 出版日期:2024-03-20 发布日期:2024-04-03

Jianpi Zishen granule inhibits podocyte autophagy in systemic lupus erythematosus: a network pharmacology and clinical study

CHEN Junjie, HUANG Chuanbing, LI Ming   

  1. First Clinical College, Anhui University of Chinese Medicine, Hefei 230000, China; Department of Rheumatology and Immunology, First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, China
  • Online:2024-03-20 Published:2024-04-03

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摘要： 目的 基于网络药理学结合分子对接，通过临床验证，探讨健脾滋肾方从足细胞自噬途径治疗系统性红斑狼疮（SLE）潜在机制。方法 运用TCMSP、GeneCards、OMIM及TTD数据库获取健脾滋肾方及SLE、足细胞自噬相关靶点；取三者交集得到健脾滋肾方经足细胞自噬途径治疗SLE的潜在作用靶点。Cytoscape3.9.1构建“复方-成分-疾病-潜在靶点”网络及蛋白质互作网络，筛选出关键活性成分和靶点，进行分子对接。并临床纳入46例SLE患者和10例健康人，将SLE患者随机分为观察组（n=22）和对照组（n=21）。对照组口服醋酸泼尼松及吗替麦考酚酯，观察组在其基础上加用健脾滋肾方，疗程均为12周。运用ELISA检测SLE组及健康人群尿液中nephrin、synaptopodin水平；使用Western blot检测外周血内p-JAK1/JAK1、p-STAT1/STAT1、LC3II/LC3I、p62蛋白水平，并观察经健脾滋肾方干预后对SLE患者足细胞相关蛋白（nephrin、synaptopodin）、自噬水平（LC3II/LC3I、p62）及p-JAK1/JAK1、p-STAT1/STAT1表达影响。结果 经筛选得到健脾滋肾方经足细胞自噬途径治疗SLE的关键活性成分4种，核心靶点基因5个（STAT1、PIK3CG、MAPK1、PRKCA、CJA1）；富集分析显示可能涉及AGE-RAGE、JAK/STAT、EGFR、PI3K/Akt等信号通路。分子对接结果显示关键活性物质和核心靶点均有较好的结合活性。其中STAT1 是结合活性最好的靶蛋白，表明STAT1 可能是健脾滋肾方给药后负责信号转导的重要介质。临床试验证实健脾滋肾方能够显著降低血清中p-JAK1/JAK1及p-STAT1/STAT1、LC3II/LC3I蛋白表达水平（P<0.05或P<0.01），升高p62蛋白表达水平（P<0.05），降低尿液中nephrin、synaptopodin水平（P<0.05）。结论 健脾滋肾方经足细胞自噬治疗SLE具有多成分、多靶点、多通路协同作用的特点，其可能通过槲皮素、山奈酚、β-谷甾醇、异鼠李素等关键活性成分作用于STAT1靶基因，抑制JAK/STAT通路相关基因表达，调控自噬减轻SLE足细胞损伤。

关键词: 系统性红斑狼疮；健脾滋肾方；网络药理学；足细胞；自噬

Abstract: Objective To explore the therapeutic mechanism of Jianpi Zishen (JPZS) granules for systemic lupus erythematosus (SLE) in light of podocyte autophagy regulation. Methods TCMSP, GeneCards, OMIM, and TTD databases were used to obtain the targets of JPZS granules, SLE, and podocyte autophagy. The protein-protein interaction network was constructed using Cytoscape, and the key active ingredients and targets were screened for molecular docking. In the clinical study, 46 patients with SLE were randomized into two groups to receive baseline treatment with prednisone acetate and mycophenolate mofetil (control group) and additional treatment with JPZS granules (observation group) for 12 weeks, with 10 healthy volunteers as the healthy control group. Urinary levels of nephrin and synaptopodin of the patients were detected with ELISA. Western blotting was performed to determine peripheral blood levels of p- JAK1/JAK1, p-STAT1/STAT1, LC3II/LC3I, and p62 proteins of the participants. Results Four key active ingredients and 5 core target genes (STAT1, PIK3CG, MAPK1, PRKCA, and CJA1) were obtained, and enrichment analysis identified the potentially involved signaling pathways including AGE-RAGE, JAK/STAT, EGFR, and PI3K/Akt. Molecular docking analysis showed that STAT1 was the most promising target protein with the highest binding activity, suggesting its role as an important mediator for signal transduction after JPZS granule treatment. In the 43 SLE patients available for analysis, treatment with JPZS granule significantly reduced serum levels of p- JAK1/JAK1, p- STAT1/STAT1, and LC3II/LC3I (P<0.05 or 0.01), increased the protein level of P62 (P<0.05), and reduced urinary levels of nephrin and synaptopodin (P<0.05). Conclusion The therapeutic effect of JPZS granules on SLE is mediated probably by coordinated actions of quercetin, kaempferol, β-sitosterol, and isorhamnetin on their target gene STAT1 to inhibit the JAK/STAT pathway, thus suppressing autophagy and alleviating podocyte injuries in SLE.

Key words: systemic lupus erythematosus; Jianpi Zishen granules; network pharmacology; podocytes; autophagy