Integrating Network Pharmacology and Experimental Validation to Explore the Key Mechanism of Gubitong Recipe in the Treatment of Osteoarthritis

Abstract

Background. Gubitong Recipe (GBT) is a prescription based on the Traditional Chinese Medicine (TCM) theory of tonifying the kidney yang and strengthening the bone. A previous multicentral randomized clinical trial has shown that GBT can effectively relieve joint pain and improve quality of life with a high safety in treating osteoarthritis (OA). This study is aimed at elucidating the active compounds, potential targets, and mechanisms of GBT for treating OA. Method. The network pharmacology method was used to predict the key active compounds, targets, and mechanisms of GBT in treating OA. An OA rat model was established with Hulth surgery, and the pathological changes of articular cartilage were observed to evaluate the effects of GBT. Chondrocytes were stimulated with LPS to establish in vitro models, and key targets and mechanisms predicted by network pharmacology were verified via qRT-PCR, ELISA, western blot, and immunofluorescence. The Contribution Index Model and molecular docking were used to determine the key active compounds of GBT and the major nodes affecting predicted pathways. Result. A total of 500 compounds were acquired from related databases, where 87 active compounds and their 254 corresponding targets were identified. 2979 OA-related genes were collected from three databases, 150 of which were GBT-regulating OA genes. The compound-target network weight analysis and PPI results showed that IL-6 and PGE2 are key targets of GBT in treating OA. KEGG results showed that PI3K/AKT, Toll-like receptor, NFκB, TNF, and HIF-1 are the key signaling pathways. An in vivo experiment showed that GBT could effectively suppress cartilage degradation of OA rats. In vitro experiments demonstrated that GBT can inhibit the key targets of KEGG-related pathways. Molecular-docking results suggested that luteolin, licochalcone A, and β-carotene were key targets of GBT, and the mechanisms may be associated with the NFκB signaling pathway. Blockage experiments showed that the NFκB pathway is the key pathway of GBT in treating OA. Conclusion. This study verified that GBT can effectively protect articular cartilage through multitarget and multipathway, and its inhibitory effect on the NFκB pathway is the most key mechanism in treating OA.

Chondroprotective Effects of Gubitong Recipe via Inhibiting Excessive Mitophagy of Chondrocytes

“…Our study showed that the GBT recipe can effectively suppress the cartilage degeneration and apoptosis of the chondrocytes in the OA rat model. This was consistent with a previous study [ 28 ]. We found an increased expression of MMP-13 in the OA rat model by immunohistochemistry.…”

Potential Effects of Traditional Chinese Medicine in Anti-Aging and Aging-Related Diseases: Current Evidence and Perspectives

“… Atherosclerosis Qing-Xue-Xiao-Zhi formula Reynoutria japonica Houtt., Rheum palmatum l., Typha angustifolia l., Curcuma longa l., Alisma plantago-aquatica Linn., Dioscorea septem loba Thunb. Artemisia capillaris Thunb., Atractylodes macrocephala Koidz., Acori tatarinowii Rhizoma, Carthamus tinctorius l. Regulate TLR4/MyD88/NF-κB pathway [ 98 ] Parkinson’s disease Yiguanjian decoction Glehniae Radix, Toosendan Fructus, Angelicae Sinensis Radix, Rehmanniae Radix Praeparata, Lycii Fructus, Ophiopogon japonicus, Regulate the PI3K-Akt pathway, modulate endocrine resistance of the central nervous [ 99 ] Osteoporosis Qing’e Pill Eucommia ulmoides Oliv., Psoralea corylifolia L., Juglans regia L., and Allium sativum L. Inhibit ferroptosis and promoting cell survival via the PI3K/AKT pathway and ATM [ 100 ] Osteoarthritis Gubitong Recipe Davallia mariesii Moore ex bak, Epimedium brevicornu Maxim., Psoralea corylifolia Linn., Eucommia ulmoides Oliver, Lycium chinense Miller, Rhizoma Bolbostematis, Stephania tetrandra S. Moore, Spatholobus suberectus Dunn Inhibit the NF κB pathway [ 101 ] Chronic kidney disease Jian-Pi-Yi-Shen Formula Astragalus membranaceus, Dioscorea opposita Thunb. Rehmannia glutinosa, Cistanche deserticola Ma, Amomum kravanh Pierre ex Gagnep.…”

Six Genes Associated with Lymphatic Metastasis in Colon Adenocarcinoma Linked to Prognostic Value and Tumor Immune Cell Infiltration

“…Gene Ontology (GO) enrichment analysis (BP: biological process; CC: cellular component; MF: molecular function) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed on the selected differential lymphoid metastasis molecules [18][19][20]. "q value <0.2 and p. adj <0.1" were used as the threshold to enrich functional categories and pathways.…”