文献类型：期刊文献

中文题名：基于网络药理学和实验验证探讨骨痨愈康丸含药血清干预类风湿关节炎骨破坏的作用机制

英文题名：Mechanism of Serum Containing Gulao Yukang(骨痨愈康)Pills in Intervening in Bone Destruction in Rheumatoid Arthritis Based on Network Pharmacology and Experimental Validation

作者：田杰祥[1];王钢[1];宋敏[2];漆文霞[2];姜刚刚[2];李平顺[1];贺童[2];魏勇[1];安文博[1]

第一作者：田杰祥

机构：[1]甘肃中医药大学附属医院,兰州730020;[2]甘肃中医药大学,兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2025

卷号：41

期号：5

起止页码：9

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金地区基金资助项目(编号:81960832);甘肃省自然科学基金项目(编号:22JR5RA588)。

语种：中文

中文关键词：骨痨愈康丸;类风湿关节炎;骨破坏;网络药理学;核因子κB信号通路

外文关键词：Gulao Yukang(骨痨愈康)Pills;Rheumatoid arthritis;Bone destruction;Network pharmacology;NF-kB signaling pathway

摘要：目的:基于网络药理学和实验验证探讨骨痨愈康丸含药血清干预类风湿关节炎骨破坏的作用机制。方法:利用中药系统药理学分析平台和中药分子机制的生物信息学分析工具筛选骨痨愈康丸组方中药活性成分及其相关靶点,筛选以类风湿关节炎骨破坏靶点的基因,构建“药物-成分-靶点-疾病”网络图。建立RAW 264.7诱导的破骨细胞模型,以骨痨愈康丸高中低剂量含药血清进行干预,CCK-8法检测RAW 264.7细胞增殖情况;ELISA法检测细胞上清液中TNF-α、MMP-3、IL-1β的含量;Western blotting法检测细胞C-FOS、NFATC1、NF-κB p65、p-NF-κB p65、IκBα、p-IκBα蛋白的表达;免疫荧光染色检测NF-κB p65核移位。结果:通过网络药理学数据库和中药分子机制的生物信息学分析工具,得到骨痨愈康丸治疗类风湿关节炎骨破坏的潜在靶点47个,活性成分184种,主要有效成分为槲皮素、银杏叶酮、β-谷甾醇,关键靶点有炎性细胞因子TNF-α、IL-17,生物学过程涉及对细胞因子及骨破坏的反应,涉及的通路主要有NF-κB、PI3K/AKT等。细胞实验结果显示,与空白对照组相比,模型对照组细胞中TNF-α、MMP-3、IL-1β含量明显升高(P<0.05),C-FOS、NFATC1、NF-κB P65、p-NF-κB P65蛋白表达上调(P<0.05),p-IκBα蛋白表达下调(P<0.05),NF-κB P65在细胞核的表达上调(P<0.05);与模型对照组相比,不同剂量的骨痨愈康丸含药血清组细胞中TNF-α、MMP-3、IL-1β含量明显降低(P<0.05);0.95、1.90 g/kg骨痨愈康丸10%含药血清组细胞中的C-FOS、NFATC1、NF-κB P65、p-NF-κB P65蛋白表达下调(P<0.05),各浓度骨痨愈康丸含药血清组细胞中p-IκBα表达上调(P<0.05),NF-κB P65(红色)在细胞核的表达下调(P<0.05)。结论:骨痨愈康丸可抑制C-FOS、NFATC1、NF-κB P65和IκB的活化和磷酸化表达,减少炎性介质分泌,抑制NF-κB P65核移位,调控NF-κB的信号传导通路对类风湿关节炎骨破坏发挥保护作用。
Objective:To explore the mechanism of action of Gulao Yukang(骨痨愈康)Pills-containing serum in intervening in bone destruction in rheumatoid arthritis based on network pharmacology and experimental validation.Methods:Traditional Chinese Medicine Systems Pharma-cology Database and Analysis Platform(TCMSP)and the bioinformatics analysis tool of molecular mechanism of Chinese medicine were used to screen the active ingredients of Chinese medicine in Gulao Yukang Pills and their related targets.Genes targeting diseases related to bone destruction in rheumatoid arthritis were screened,and a diagram of"drug-ingredient-target-disease"network was constructed.The model was established in RAW 264.7-induced osteoclasts,and the low-,medium-,and high-dose Gulao Yukang Pills-containing serum was used for inter-vention.The proliferation of RAW 264.7 cells was detected by CCK-8 assay,and the content of TNF-α,MMP-3,and IL-1β was detected by ELISA in the supernatant of the cells.The protein expression of cellular C-FOS,NFATC1,NF-κB p65,p-NF-κB p65,IκBα,and p-IκBα was detected by Western blotting,and immunofluorescence staining was used to detect NF-κB p65 nuclear translocation.Results:Through the net-work pharmacology database and the bioinformatics analysis tool of the molecular mechanism of Chinese medicine,47 potential targets and 184 active ingredients of Gulao Yukang Pills for the treatment of bone destruction in rheumatoid arthritis were obtained,involving main active in-gredients quercetin,Ginkgo biloba ketone,and lysine β-sitosterol.Key targets were cellular inflammatory factors TNF and IL-17.The biologi-cal processes involved the response to cytokines and bone destruction,and the main pathways involved NF-κB,PI3K/AKT,etc.Based on the results of cell experiments,compared with the results in the blank control group,the content of TNF-α,MMP-3,and IL-1β was significantly in-creased in the model control group(P<0.05),and the protein expression levels of C-FOS,NFATC1,NF-κB p65,and p-NF-κB p65 were up-regulated(P<0.05).Moreover,the protein expression of p-IκBα was downregulated(P<0.05),and the expression of NF-κB p65 in the cell nucleus was upregulated(P<0.05).Compared with the results in the model control group,the cell content of TNF-α,MMP-3,and IL-1β in Gulao Yukang Pills-containing serum groups with different doses was significantly decreased(P<0.05),and the protein expression levels of C-FOS,NFATC1,NF-κB p65,and p-NF-κB p65 in the cells of 10%drug-containing serum groups at doses of 0.95 g/kg and 1.90 g/kg were downregulated(P<0.05).In Gulao Yukang Pills-containing serum groups with different doses,the p-IκBα expression was upregulated(P<0.05),while the expression of NF-κB p65(red)in the cell nucleus was downregulated(P<0.05).Conclusions:Gulao Yukang Pills can inhibit the activation and phosphorylation expression of C-FOS,NFATC1,NF-κB p65,and IκBα,reduce the secretion of inflammatory media-tors,inhibit the nuclear translocation of NF-κB p65,and regulate the NF-κB signaling pathway to play a protective role in bone destruction in rheumatoid arthritis.