Senomorphic agent pterostilbene ameliorates osteoarthritis through the PI3K/AKT/NF-κB axis: an in vitro and in vivo study.

ABSTRACT

OBJECTIVES: Osteoarthritis (OA) is the most common joint disease in the world. Among the many risk factors for OA, aging is one of the most critical factors. The treatment with senop-associated secretory phenotype (SASP) is one of the important, promising anti-aging strategies at present. Pterostilbene (PTE) is a trans-stilbene compound with anti-tumor, anti-oxidation, anti-inflammatory, and anti-aging pharmacologic activities. The purpose of this study is to explore the therapeutic effects of PTE on articular chondrocyte senescence and OA and its related mechanisms. METHODS: Male Sprague-Dawley rats were operated on with transection of the anterior cruciate ligament (ACLT) and a destabilized medial meniscus (DMM) surgery to establish the OA model and then injected intraperitoneally with PTE (20 mg/kg) for 5 weeks. Finally, rats were sacrificed and knee joints were collected for histologic analysis. Rat chondrocytes were stimulated with interleukin-1ß (IL-1ß) with or without PTE treatment. The therapeutic effects of PTE and related mechanisms were investigated by examining and analyzing relative markers through senescence-associated ß-galactosidase (SA-ß-Gal) assay, cell cycle, qRT-PCR, western blot, bioinformatic analysis, immunofluorescence, and molecular modeling. RESULTS: With in vivo experiments, PTE can significantly reduce the Mankin scores and OARSI scores of the knee joint in ACLT+DMM OA model rats and reduce the interleukin-6 (IL-6) level in the knee lavage fluid. Immunohistochemical staining showed that compared to the OA group, the PTE treatment group had significantly increased expression of collagen type II in articular cartilage, and significantly decreased matrix metalloproteinase 13 (MMP-13) and IL-6, the main SASP proteins, and had expression of p16 and p21, markers of aging in chondrocytes. In vitro, PTE reduced the ratio of SA-ß-Gal positive chondrocytes and G0-G1 phase chondrocytes in IL-1ß-induced rat chondrocytes. PTE significantly inhibited the expression of MMP-13, IL-6, thrombospondin motif 5 (ADAMTS5), p16, and p21, and significantly increased the expression of collagen type II. Bioassay and subsequent western blot showed that PTE significantly inhibited the activation of PI3K/AKT and NF-κB signaling pathways. The results of molecular docking experiments showed that PTE could bind closely to the sites of PI3K protein, thereby inhibiting the phosphorylation of PI3K. CONCLUSIONS: The experimental results indicate that PTE plays an anti-chondrocyte senescence role in the treatment of OA by inhibiting the PI3K/AKT/NF-κB signaling pathway and reducing expression of SASP.