基于网络药理学探讨莱菔及莱菔子防治痛风性关节炎的作用机制  

Exploring the mechanism of action of Raphanus sativus L and Raphani semen against gouty arthritis based on network pharmacology

作　　者：许诺 高明利[2] 赵夜雨[2] 关蕊[1] 齐庆[2] 孙蓬远[2] 张瑞君 XU Nuo;GAO Mingli;ZHAO Yeyu;GUAN Rui;QI Qing;SUN Pengyuan;ZHANG Ruijun(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Liaoning University of Chinese Medicine Affiliated Hospital,Shenyang 110032,China;Shenyang Kangfenghong Biotechnology Co.,Shenyang 110001,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032 [3]沈阳抗风竑生物技术有限公司,辽宁沈阳110001

出　　处：《沈阳药科大学学报》2025年第1期74-85,共12页Journal of Shenyang Pharmaceutical University

基　　金：辽宁省科学技术计划项目(2022-PJLH-04)。

摘　　要：目的采用网络药理学与分子对接探讨莱菔及莱菔子防治痛风性关节炎(gouty arthritis,GA)的作用机制,并建立急性痛风性关节炎(acute gouty arthritis,AGA)大鼠模型进行验证。方法通过TCMSP、HERB数据库及文献查阅获得两味药潜在活性成分与治疗靶点,通过Genecards、OMIM、DisGeNET数据库获得GA靶点,利用VENNY2.1平台计算得出莱菔及莱菔子成分与GA的交集靶点。对交集靶点进行GO功能及KEGG通路富集分析。雄性SD大鼠随机分为空白组、模型组、雌性红莱菔(0.27 g·kg^(-1))+莱菔子(0.27 g·kg^(-1))组、美洛昔康组(0.66 mg·kg^(-1)),每组8只,踝关节注射单钠尿酸盐(monosodiumurate,MSU)混悬液建立AGA大鼠模型。采用游标卡尺测量大鼠踝关节直径,计算踝关节肿胀度;采用苏木素-伊红(HE)染色观察踝关节滑膜组织病理变化;采用ELISA、Western blotting检测踝关节滑膜组织中关键靶点和信号通路的表达。结果网络药理学共筛选出莱菔及莱菔子的10种活性成分及262个潜在靶点,GA相关靶点1949个,交集靶点87个,其中核心靶点13个,富集分析得出TNF通路、PI3K/AKT通路及癌症相关通路等,TNF通路是莱菔及莱菔子干预GA最有可能性的通路。大鼠踝关节注射MSU混悬液后明显肿胀(P<0.001),滑膜细胞坏死脱落,结缔组织重度增生,大量炎性细胞浸润,NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、半胱天冬酶-1(cysteinyl aspartate specific proteinase-1,caspase-1)、消皮素D(gasdermin D,GSDMD)、IL-1β、IL-18及TNF-α表达水平明显升高(P<0.001、P<0.01);雌性红莱菔及美洛昔康治疗后,有效缓解关节肿胀(P<0.001、P<0.01、P<0.05),关节滑膜坏死、炎性细胞浸润明显改善,同时显著抑制IL-1β、IL-18、TNF-α水平及NLRP3、ASC、Caspase-1、GSDMD蛋白表达(P<0.001、P<0.01�Objective Network pharmacology and molecular docking were used to explore the mechanism of action of Raphanus sativus L and Raphani semen against gouty arthritis(GA),and a rat model of acute gouty arthritis(AGA)was established for validation.Methods The potential active ingredients and therapeutic targets of the two herbs were obtained from TCMSP,HERB database and published literatures,and the GA targets were obtained from Genecards,OMIM and DisGeNET databases,and the intersecting targets of Raphanus sativus L and Raphani semen constituents with GA were calculated by using VENNY2.1 platform.The intersecting targets were analyzed for GO function and KEGG pathway enrichment.Male SD rats were randomly divided into blank group,model group,female red radish(0.27 g·kg^(-1))+radish semen(0.27 g·kg^(-1))group,meloxicam group(0.66 mg·kg^(-1)),8 rats in each group,and ankle joint injection of MSU suspension was used to establish AGA rat model.Vernier calipers were used to measure the diameter of the ankle joints of rats and calculate the swelling degree of the ankle joints;Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of the synovial tissue of the ankle joints;ELISA and Western blotting were used to detect the expression of key targets and signaling pathways in the synovial tissue of the ankle joints.Results A total of 10 active components and 262 potential targets of Raphanus sativus L and Raphani semen were screened in the network pharmacology,and 1949 GA-related targets and 87 intersecting targets were identified,among which 13 were core targets.Enrichment analysis showed that TNF pathway,PI3K/AKT pathway and cancer-related pathway were identified,and that TNF pathway was the most probable pathway of Raphanus sativus L and Raphani semen for interfering with GA.The ankle joints of rats injected with MSU suspension were significantly swollen(P<0.001),with necrotic detachment of synovial cells,heavy proliferation of connective tissues,and infiltration of a large number of inflammatory c

关 键 词：网络药理学 莱菔 莱菔子 痛风性关节炎 分子机制 

分 类 号：R96[医药卫生—药理学]