Harpagide inhibits the TNF--induced inflammatory response in rat articular chondrocytes by the glycolytic pathways for alleviating osteoarthritis

Osteoarthritis (OA) causes severe and functional dysfunction due to abnormal inflammation. The objective of this study was to evaluate the effect of Harpagide (HPG) on TNF-alpha-induced inflammation in vitro and in vivo. The effect of HPG on the proliferation of rat chondrocytes was studied. The anti-inflammatory effect of HPG and its molecular mechanisms were elucidated by qPCR, Western blotting, flow cytometry, metabolome analysis in vitro. In addition, the OA rat model was established, and the effect of HPG on OA was verified in vivo. We revealed 10 mu M HPG demonstrated biocompatibility. The results demonstrated that HPG restored the upregulation of MMP-13, COX2, IL-1 beta and IL-6 induced by TNF-alpha. Moreover, HPG reversed TNF-alpha induced degradation of the extracellular matrix of chondrocytes. TNF-alpha treatment induced down-regulation of the mRNA/protein levels of proliferative markers Bcl2, CDK1 and Cyclin D1 were also recovered. HPG can inhibit TNF-alpha-induced inflammatory response through glycolytic metabolic pathways. HPG can restore TNF-alpha-induced upregulation of GRP78/IRE1 alpha, and downregulation of AMPK proteins. In vivo experiments demonstrated that after HPG treatment, the appearance and physiological structure of articular cartilage were more integrated with highly organized chondrocytes and rich cartilage matrix compared with OA group. Finally, the molecular docking of HPG and selected key factors in glycolysis results showed that HPG had good binding potential with PFKM, PFKP, PFKFB3, PKM, HK2, and PFKL. In conclusion, the results shown HPG protects and activates chondrocytes, inhibits TNF-alpha-induced inflammatory response by glycolysis pathway in rat articular chondrocytes, and plays a role in the treatment of OA.