中国药物警戒 ›› 2026, Vol. 23 ›› Issue (3): 241-249.
DOI: 10.19803/j.1672-8629.20250638

• 院士笔谈 • 上一篇    下一篇

基于蛋白质组学解析雷公藤红素诱导成纤维细胞转分化的机制

孙悦1,2, 张宇男2,3△, 孙源伯2,4, 李高芾2, 杨芳1, 王宁宁2, 张鹏飞2, 高月2#, 周维1,2,*   

  1. 1青海大学药学院,青海 西宁 810016;
    2军事科学院军事医学研究院,北京 100850;
    3北京中医药大学生命科学学院,北京 102488;
    4清华大学自动化系,北京 100084
  • 收稿日期:2025-09-09 出版日期:2026-03-15 发布日期:2026-03-17
  • 通讯作者: *周维,男,博士,副研究员,中药药理与毒理研究。E-mail: zhouweisyl802@163.com。#为共同通信作者。
  • 作者简介:孙悦,女,在读硕士,药学。△为并列第一作者。高月,中国工程院院士,军事药学和中药药理学专家,专业技术少将军衔,中国医学科学院学术咨询委员会委员。现为全军特需中药及天然药物重点实验室主任。先后主持国家自然科学基金重大项目、国家重点研发计划等23项。以第一完成人获得国家科技进步一等奖、二等奖和军队科技进步一等奖等6项; 荣获何梁何利基金科学与技术进步奖、首批全国创新争先奖状、全国中医药杰出贡献奖、树兰医学奖等。荣立个人二等功2次、三等功1次。
  • 基金资助:
    国家自然科学基金资助项目（82192910）

Proteomics-Based Analysis of Mechanisma Underlying Celastrol-Induced Fibroblast Transdifferentiation

SUN Yue1,2, ZHANG Yunan2,3△, SUN Yuanbo2,4, LI Gaofu2, YANG Fang1, WANG Ningning2, ZHANG Pengfei2, GAO Yue2#, ZHOU Wei1,2,*   

  1. 1School of Pharmacy, Qinghai University, Xining Qinghai 810016, China;
    2Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China;
    3School of Life Sciences, Beijing University of Chinese Medicine, Beijing 102488, China;
    4Department of Automation, Tsinghua University, Beijing 100084, China
  • Received:2025-09-09 Online:2026-03-15 Published:2026-03-17

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摘要： 目的 通过蛋白质组学分析探讨雷公藤红素产生心脏毒性及潜在促进纤维化的机制。方法 提取乳鼠原代心脏成纤维细胞,利用Cell Counting Kit-8（CCK-8）实验进行雷公藤红素给药浓度及时间的确定,观察成纤维细胞给药前后细胞形态的变化。雷公藤红素给药后（72 h,0.01 μmol·L-1）,采用实时荧光定量PCR测定相关促进纤维化基因表达水平;酶联免疫吸附试验（ELISA）测定细胞外基质（Extracellular Matrix,ECM）金属蛋白酶抑制剂-1（TIMP-1）、基质金属蛋白酶-2（MMP2）的含量变化;免疫荧光实验测定相关纤维化蛋白表达情况;通过蛋白质组学探讨雷公藤红素潜在促进纤维化机制。结果 雷公藤红素给药后（72 h,0.01 μmol·L-1）,与空白对照组相比,给药组细胞形态由梭形向多边形或扁平铺展状转变,且细胞体积增大。雷公藤红素给药组促纤维化相关基因及蛋白的表达水平明显升高,且细胞外基质TIMP-1、MMP2含量升高。基于蛋白质组学的差异蛋白富集分析表明,雷公藤红素影响成纤维细胞中纤维连接蛋白1（Fn1）、整合素α11（Itga11）、整合素β3（Itgb3）、转化生长因子β1诱导蛋白1（Tgfb1i1）等蛋白的表达,通过激活炎症与细胞外基质相关通路,进而促进心脏纤维化。结论 短期雷公藤红素给药对心脏功能产生潜在影响,其可能通过影响胶原蛋白合成和干扰ECM平衡,促进心脏成纤维细胞向肌成纤维的转变,加剧纤维化进程。

关键词: 雷公藤红素, 成纤维细胞, 转分化, 纤维化, 细胞外基质, 蛋白质组学, 乳鼠

Abstract: Objective To explore the mechanisms of celastrol-induced cardiotoxicity and its potential promotion of fibrosis via proteomic analysis. Methods Primary cardiac fibroblasts were isolated from neonatal mice. The Cell Counting Kit-8 (CCK-8) assay was used to determine the concentration and duration of celastrol treatment. Bright-field microscopy was employed to observe the morphological changes of fibroblasts before and after treatment. Seventy-two hours after treatment with 0.01 μmol·L-1, real-time quantitative PCR (qPCR) was used to measure the expression levels of related pro-fibrotic genes. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify changes in the levels of extracellular matrix (ECM), including tissue inhibitor of metalloproteinases-1 (TIMP-1) and matrix metalloproteinase-2 (MMP-2). Immunofluorescence staining was performed to calculate the expressions of fibrosis-related proteins. Proteomic analysis was used to investigate the potential mechanisms by which celastrol promoted fibrosis. Results 72 hours after treatment with celastrol 0.01μmol·L-1, the morphology of cells in the treatment group changed from spindle-shaped to polygonal or flattened, and the cell volume increased compared with the blank control group. The expression levels of pro-fibrotic genes and proteins were significantly elevated in the celastrol-treated group, along with increased levels of the extracellular matrix proteins TIMP-1 and MMP2. Differential protein enrichment analysis based on proteomics indicated that celastrol affected the expressions of such proteins as fibronectin 1 (Fn1), integrin alpha 11 (Itga11), integrin beta 3 (Itgb3), and transforming growth factor beta 1 induced transcript 1 (Tgfb1i1) in fibroblasts. By activating inflammatory and extracellular matrix-related pathways, celastrol subsequently promoted cardiac fibrosis. Conclusion Short-term administration of celastrol may potentially impact cardiac function, likely by influencing collagen synthesis and disrupting extracellular matrix (ECM) homeostasis, thereby promoting the transformation of cardiac fibroblasts into myofibroblasts and exacerbating the fibrosis process.

Key words: Celastrol, Fibroblast, Transdifferentiation, Fibrosis, Extracellular Matrix (ECM), Poteomics, Neonatal Mice

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