基于网络药理学和细胞实验探讨独活—牛膝治疗骨关节炎的作用机制  

Mechanism of Achranthis Bioentatae Radix-Angelicae Pubescentis Radix in the treatment of osteoarthritis based on network pharmacology and molecular docking

作　　者：庄耀隆 闵令田 ZHUANG Yaolong;MIN Lingtian(Suzhou Hospital of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Suzhou,Jiangsu 215009,China;Nantong Hospital of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing University of Chinese Medicine,Nantong,Jiangsu 226000)

机构地区：[1]南京中医药大学附属苏州市中医医院,江苏苏州215009 [2]南京中医药大学附属南通市中医院,江苏南通226000

出　　处：《徐州医科大学学报》2023年第9期650-656,共7页Journal of Xuzhou Medical University

基　　金：南通市科技局指导性项目(JCZ20195)。

摘　　要：目的探讨独活和牛膝治疗骨性关节炎(OA)的作用机制。方法通过网络药理学获得了OA相关靶标,构建了中药—活性成分—蛋白网络、蛋白—蛋白互作网络和核心基因网络,并对筛选得到的核心基因进行京都基因和基因组百科全书(KEGG)功能分析。细胞实验采用白细胞介素-1β(IL-1β)诱导的OA细胞模型,采用CCK8检测药物对于细胞活力的影响,Western blot及RT-PCR检测通路蛋白及基因的表达。结果独活—牛膝药对共得到14个活性化合物和35个潜在OA靶点,15个核心基因。其中血管内皮生长因子A(VEGFA)-肿瘤蛋白53(TP53)是独活—牛膝的网络和通路中的重要节点,也是可能是独活—牛膝治疗骨性关节炎的关键通路。另外,通过人类OA样本的转录组数据分析表明,VEGFA和TP53在OA患者软骨中显著降低,同时2个基因相关性显著。IL-1β诱导的OA细胞模型表明独活—牛膝可以显著激活VEGFA-TP53通路,同时显著降低MMP13基因和蛋白表达并上调collagenⅡ的基因和蛋白表达。结论独活—牛膝可能主要通过上调VEGFA-TP53通路从而影响其他多条通路,降低了OA发展过程中的关键蛋白,从而发挥其抗OA的功效。Objective To explore the mechanism of Achranthis Bioentatae Radix(AB)and Angelicae Pubescentis Radix(AP)in the treatment of osteoarthritis(OA).Methods In this study,the OA-related targets were screened out through network pharmacology.Then,a traditional Chinese medicine-active ingredient-protein network,a protein-protein interaction network and a core gene network were established.The resultant core genes were applied for KEGG function analysis.Furthermore,an interleukin(IL)-1β-induced model of OA cells was constructed.Cell viability was detected by CCK-8 assay.The expression of signaling proteins and gens were detected by Western blot and RT-PCR.Results A total of 14 active compounds,35 potential OA targets,and 15 core genes were obtained from the drug pair of AB-AP.Among them,vascular endothelial growth factor A(VEGFA)and tumor protein 53(TP53)were important nodes in the network and pathways of AB-AP,and might also be a key pathway for AB-AP to treat OA.Furthermore,the transcriptome data analysis of human OA samples showed that VEGFA and TP53 significantly decreased in the cartilage of OA patients,and the two genes were significantly correlated.The IL-1β-induced OA cell model showed that AB-AP significantly activated the VEGFA-TP53 pathway,while significantly reducing the gene and protein expression of MMP13 and up-regulating collagenⅡgene and protein expression.Conclusions AB-AP may mainly up-regulate the VEGFA-TP53 pathway,in order to affect other multiple pathways,reduce the key proteins in the development of OA,and exert its anti-OA effect.

关 键 词：网络药理学 体外实验 牛膝 骨性关节炎 血管内皮生长因子A 肿瘤蛋白53 

分 类 号：R593.22[医药卫生—内科学]