主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2021, Vol. 44 ›› Issue (4): 340-349.doi: 10.3969/j.issn.1006-2157.2021.04.009

• 中药药理 • 上一篇    下一篇

应用系统药理学探讨穿龙汤治疗风湿性关节炎的多层次作用机制*

江利1,2, 梁晟楠3, 王海清4, 郭子湖2,4, 王永华1,2,4#   

  1. 1 石河子大学新疆植物药资源利用教育部重点实验室 新疆 832000;
    2 陕西秦岭七药协同创新中心;
    3 西北农林科技大学化学与药学院;
    4 西北大学生命科学学院西部资源生物与生物技术教育部重点实验室
  • 收稿日期:2020-09-29 发布日期:2021-04-29
  • 通讯作者: #王永华,男,博士,教授,博士生导师,主要研究方向:中西医结合治疗心血管病的临床基础研究,E-mail:dcpwyh@163.com
  • 作者简介:江利,女,在读硕士生
  • 基金资助:
    *国家自然科学基金项目(No.U1603285),国家科技重大专项(No.2019ZX09201004-001)

Multi-level mechanisms of Chuanlong decoction in treating rheumatic arthritis from the perspective of systems pharmacology*

Jiang Li1,2, Liang Shengnan3, Wang Haiqing4, GuoZihu2,4, Wang Yonghua1,2,4#   

  1. 1 Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, Shihezi University, Xinjiang 832000, China;
    2 Collaborative Innovation Center of Qiyao in Mt. Qinling, Shaanxi 712100, China;
    3 School of Chemistry & Pharmacy, Northwest A & F University, Shaanxi 712100, China;
    4 Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Life Sciences, Northwest University, Shaanxi 712100, China
  • Received:2020-09-29 Published:2021-04-29
  • Contact: Prof. Wang Yonghua, Ph. D., Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, Shihezi University, Xinjiang 832002, E-mail: dcpwyh@163.com
  • Supported by:
    National Natural Science Foundation of China (No.U1603285); National Science and Technology Major Project (No. 2019ZX09201004-001)
    Ethical review: Committee of Ethic Review on Medical and Experimental Animals in Jiangsu Kanion Pharmaceutical Co.Ltd. (No. 2020072003)

摘要: 目的 研究极具地域特色的秦岭七药之穿山龙为主组成的穿龙汤复方对风湿性关节炎的疗效及其作用机制。方法 利用中药系统药理学数据库和分析平台(TCMSP)筛选穿龙汤各味草药中候选活性成分。利用系统打靶(SysDT)和加权系综相似度算法(WES)鉴定各味中药的靶点,并构建分子-靶标网络。进一步,整合网络分析和功能富集分析工具揭示这些潜在药理学靶点所涉及的生物过程和通路。选取穿龙汤中的主要活性成分β-谷甾醇进行小鼠体内实验验证,采用完全弗氏佐剂(0.02 mL/只)进行慢性炎症模型构建。随机将Balb/c小鼠分为正常组、模型组、β-谷甾醇组(35 mg/kg)、地塞米松组(15 mg/kg),每组6只,连续灌胃给药7 d。利用HE染色观察各组滑膜炎性程度变化;RT-PCR方法检测小鼠踝关节肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)mRNA的表达。结果 筛选出穿龙汤中候选活性成分27个和药理学靶点148个。整合网络分析和富集分析结果显示,穿龙汤中的主要活性成分薯蓣皂苷类、β-谷甾醇、川牛膝甾酮类、忍冬苷、金粉蕨素等可通过调节PI3K-AKT通路、TNF通路、Toll样受体通路等达到调节骨细胞增殖与凋亡、抗炎和免疫抑制等作用,从而达到治疗风湿性关节炎的效果。验证结果显示,模型组滑膜层增厚,炎性浸润增加,予以β-谷甾醇及地塞米松治疗后,小鼠滑膜炎性程度有明显改善,且TNF-α、IL-6的水平较模型组降低(P<0.05)。这与系统药理学预测结果基本一致,表明系统药理学预测结果具有较高的准确度。结论 穿龙汤复方中不同草药在风湿性关节炎治疗的作用机制上各有侧重,尤其是与TNF-α和Toll样受体信号通路有关。

关键词: 秦岭七药, 穿龙汤, 系统药理学, 关节炎, 中药配伍, 验证, 小鼠

Abstract: Objective To study the effects and mechanism of Chuanlong Decoction (CLT), a formula containing Chuanshanlong (Rhizoma Dioscorae Nipponicae) as the major ingredient and also a regional featured herb in Mt. Qingling area. Methods The candidate active ingredients of each herb in CLT Decoction were screened by using the Traditional Chinese Medicine System Pharmacology (TCMSP). Potential targets of these compounds were identified with System Drug Target Model (SysDT) and Weighted Ensemble Similarity (WES); a compound-target network was subsequently constructed. Further, network analysis and functional enrichment analysis were used to uncover the biological processes and signal pathways involved in these pharmacological targets. β-sitosterol, the main active ingredient in CLT Decoction, was selected to test in mice in vivo. Chronic inflammation model was established by using Freund's complete adjuvant (0.02 mL/per mouse). Mice were randomly divided into four groups: normal group, model group, β-sitosterol group and dexamethasone group (n=6 in each group for 7-day continuous intervention). The degree of synovial inflammation was measured with HE staining; The mRNA expressions of TNF-α and IL-6 in synovial tissue were detected by using RT-PCR. Results 27 candidate active ingredients in CLT Decoction were screened, and 148 pharmacological targets of arthritis were predicted. Network analysis and enrichment analysis revealed that the main active ingredients in CLT Decoction including diosgenin, β-sitosterol, cyasterone, lonicerin and onychin can regulate bone cellular proliferation, apoptosis, anti-inflammatory and immunosuppression processes via PI3K-AKT, TNF and Toll-like receptor signaling pathways. The results showed that the synovial layer significantly thickened, and that the inflammatory infiltration increased in the model group. After treatment with β-sitosterol and dexamethasone, the severity of synovial inflammation were significantly reduced, and the levels of TNF-α and IL-6 were also reduced compared with the model group (P<0.05). These results were consistent with the system pharmacological prediction results, indicating that the system pharmacological prediction was quite accurate. Conclusion Different herbal medicines in CLT Decoction may have played different roles in mechanism of action in the treatment of RA, mostly involved in TNF-α and Toll-like signaling pathways.

Key words: Qiyao in Mt. Qinling, Chuanlong Decoction, systems pharmacology, arthritis, compatibility of Chinese medicine, verification, mice

中图分类号: 

  • R285.5