Triptolide in the treatment of osteoarthritis: network pharmacology analysis and animal model validation

doi:10.12307/2026.549

Abstract

Abstract: BACKGROUND: Osteoarthritis is a chronic degenerative disease of the joints that can lead to disability. Its main pathological features are persistent inflammation and cartilage destruction. Triptolide has been used to treat a variety of chronic joint diseases. However, the mechanism of triptolide in the treatment of osteoarthritis has not been clarified
OBJECTIVE: To identify the effective targets of triptolide in the treatment of osteoarthritis by network pharmacology, and to investigate the therapeutic effect of triptolide on osteoarthritis in the osteoarthritis model.
METHODS: Network pharmacology was used to anticipate the potential targets and signaling pathways of triptolide in the treatment of osteoarthritis, and molecular docking technology was used to validate the core targets. A rat osteoarthritis model was established by anterior cruciate ligament transection. Eight weeks after modeling, the rats were administered with triptolide and sodium hyaluronate by intra-articular injection for 6 weeks. After 6 weeks of intervention, the pathological changes in rat knee joints were observed by hematoxylin-eosin staining and safranin O-fast green staining. The levels of inflammatory factors in rat serum were detected by enzyme-linked immunosorbent assay. The expression of aggrecan, type I platelet-responsive protein-containing desmoglein metalloproteinase 5, type II collagen and matrix metalloproteinase 13 proteins in rat articular cartilage was tested by immunohistochemical staining.
RESULTS AND CONCLUSION: (1) The results of network pharmacology indicated that the target of triptolide may be related to the inhibition of the release of factors such as interleukin 6, tumor necrosis factor ɑ, interleukin 1β, matrix metalloproteinase 9, and the over-activation of the nuclear factor-κB/JAK2-STAT3 signaling pathway. (2) Triptplide could reduce the degree of joint swelling in osteoarthritic rats; pathologically improve the articular cartilage and maintain the cartilage structure; decrease the serum levels of interleukin 6, tumor necrosis factor ɑ, interleukin 1β, matrix metalloproteinase 9, and matrix metalloproteinase 3 in osteoarthritic rats; reduce the protein expression of matrix metalloproteinase 13 and type I platelet-responsive protein-containing desmoglein metalloproteinase 5 in the articular cartilage; and increase the expression of type II collagen and aggrecan in the cartilage, thereby achieving cartilage protection.

Key words: osteoarthritis, triptolide, network pharmacology, inflammation, articular cartilage, matrix metalloproteinase 13, nuclear factor-κB signaling pathway, engineered tissue construction

CLC Number:

Chen Yixian, Chen Chen, Lu Liheng, Tang Jinpeng, Yu Xiaowei. Triptolide in the treatment of osteoarthritis: network pharmacology analysis and animal model validation[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(4): 805-815.

Figures/Tables 11

2.2 蛋白相互作用网络图的构建及核心靶点的筛选将交集靶点导入STRING数据库获得靶点基因对应的蛋白互作网络，见图2A。通过Cytoscape软件将蛋白互作网络进行可视化，得到67个节点和1 084条边，见图2B。采用CytoNCA插件对度中心性、介数中心性、接近中心性、特征向量中心性、基于局部平均连通性的方法和网络中心性进行参数运算以筛选核心靶基因，运算范围为分别大于6个参数的中位数，循环运算2次。最终获得14个核心基因，分别为 NFKB1、MMP9、IL6、TNF、IL1β、TP53、STAT3、BCL2、MYC、PTEN、JUN、PTGS2、CASP3、IFNG，见图3。说明雷公藤甲素治疗骨关节炎与细胞炎症、细胞凋亡等关系密切。 2.3 GO及KEGG富集分析通过DAVID数据库获得GO功能400条(P < 0.05)，其中生物过程325条、细胞组分29条、分子功能46条。选取排名前10的条目借助微生信平台进行可视化，生成GO功能条形图，见图4。雷公藤甲素可能通过调控细胞凋亡、炎症反应、细胞因子活性、基因表达、聚合酶Ⅱ启动子的转录等方式发挥治疗骨关节炎作用。KEGG富集分析共获得123条通路(P < 0.05)，选取其中具有代表性的25条通路进行可视化分析，见图5。雷公藤甲素治疗骨关节炎主要涉及核因子κB(nuclear factor κB，NF-κB)信号通路、Janus激酶信号转导和转录激活因子(janus kinase/signal transducer and activator of transcription，JAK/STAT)信号通路、丝裂原活化蛋白激酶(mitogen-activated protein kinase，MAPK)信号通路、白细胞介素17信号通路、肿瘤坏死因子(tumor necrosis factor，TNF)信号通路、Toll样受体通路、核苷酸结合寡聚化域(nucleotide-binding oligomerization domain，NOD)样受体通路、缺氧诱导因子1(hypoxia inducible factor-1，HIF-1)通路、P53通路、T细胞受体通路、C型凝集素受体信号通路等。 2.4 分子对接结果运用AutoDockTools软件对雷公藤甲素与核心靶点NFKB1、MMP9、IL6、TNF、IL1β进行分子对接，结果显示分子结合能均≤-29.29 kJ/mol，说明雷公藤甲素与骨关节炎的核心靶点具有较好的结合作用，见表5。最后通过PyMOL软件，将对接结果进行可视化分析，见图6。"

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