MicroRNA‐127‐5p Regulates Matrix Metalloproteinase 13 Expression and Interleukin‐1β–Induced Catabolic Effects in Human Chondrocytes

Abstract

Objective. MicroRNAs (miRNAs), small noncoding RNA molecules, are involved in the pathogenesis of various diseases such as cancer and arthritis. The aim of this study was to determine whether miR-127-5p regulates interleukin-1␤ (IL-1␤)-induced expression of matrix metalloproteinase 13 (MMP-13) and other catabolic factors in human chondrocytes.Methods. Expression of miR-127-5p and MMP-13 by normal and osteoarthritic (OA) human cartilage was determined using real-time polymerase chain reaction. The effect of miR-127-5p on MMP-13 expression was evaluated using transient transfection of human chondrocytes or chondrogenic SW-1353 cells with miR-127-5p or its antisense inhibitor (anti-miR-127-5p). MMP-13 protein production was quantified by enzymelinked immunosorbent assay, and the involvement of miR-127-5p in IL-1␤-mediated catabolic effects was examined by immunoblotting. MicroRNA-127-5p binding with the putative site in the 3 -untranslated region (3 -UTR) of MMP-13 messenger RNA (mRNA) was validated by luciferase reporter assay.Results. There was a significant reduction in miR-127-5p expression in OA cartilage compared with normal cartilage. Up-regulation of MMP-13 expression by IL-1␤ was correlated with down-regulation of miR-127-5p expression in human chondrocytes. MicroRNA-127-5p suppressed IL-1␤-induced MMP-13 production as well as the activity of a reporter construct containing the 3 -UTR of human MMP-13 mRNA. In addition, mutation of the miR-127-5p binding site in the 3 -UTR of MMP-13 mRNA abolished miR-127-5p-mediated repression of reporter activity. Conversely, treatment with anti-miR-127-5p remarkably increased reporter activity and MMP-13 production. Interestingly, the IL-1␤-induced activation of JNK, p38, and NF-B and expression of MMP-1 and cyclooxygenase 2 were significantly inhibited by miR-127-5p. Conclusion. MicroRNA-127-5p is an important regulator of MMP-13 in human chondrocytes and may contribute to the development of OA.Osteoarthritis (OA) is a degenerative disease of articular cartilage characterized by loss of the cartilage matrix, mainly collagen and proteoglycans, leading to tissue destruction and loss of joint function. Although OA is regarded as a noninflammatory form of arthritis, considerable evidence suggests that proinflammatory cytokines derived from the synovium and chondrocytes play a role in cartilage destruction. The activity of the proinflammatory cytokine interleukin-1␤ (IL-1␤) and its downstream mediators leads to up-regulation of matrix metalloproteinase (MMP) and a decrease in the synthesis of the cartilage extracellular matrix (ECM) (1). Among the target mediators of IL-1␤, MMP-13 has gained the most interest, due to its capacity to degrade collagens along with a wide range of matrix molecules (2).A variety of therapeutic strategies for OA have thus been developed to antagonize the activity of MMP-13; however, the toxicity of nonspecific MMP inhibitors has hampered their application in clinical settings. A detailed analysis of the mechanism of regulation of MMP-13 in chondro...

Onset and Progression of Human Osteoarthritis—Can Growth Factors, Inflammatory Cytokines, or Differential miRNA Expression Concomitantly Induce Proliferation, ECM Degradation, and Inflammation in Articular Cartilage?

“…MiR-127 expression is reduced in knee OA AC compared to normal AC [190,191]. Increased MMP-13 expression upon rIL-1β treatment in monolayer cell cultures of human OA AC correlated with miR-127 suppression, with MMP-13 as a direct target of miR-127.…”

“…Increased MMP-13 expression upon rIL-1β treatment in monolayer cell cultures of human OA AC correlated with miR-127 suppression, with MMP-13 as a direct target of miR-127. In addition, miR-127 inhibits also MMP-1 expression in response to rIL-1β [190]. Collectively, with downregulation of miR-127 in OA AC, another anti-catabolic miRNA in AC is disenabled.…”

MMP-13 is constitutively produced in human chondrocytes and co-endocytosed with ADAMTS-5 and TIMP-3 by the endocytic receptor LRP1

“…The activity of MMP-13 can be regulated at multiple levels, which include transcriptional regulation [26], epigenetic modification [27], post-transcriptional regulation by microRNAs [28], [29], [30], activation of pro-enzymes [9], [10] and inhibition by endogenous inhibitors [9], [10]. In addition, Partridge and co-workers have shown that rat MMP-13 is endocytosed and degraded by osteoblastic cells [31] and this requires a two-step processes, involving a first binding to a 170 kDa cell surface receptor with a high-affinity and it is subsequently endocytosed through low-density lipoprotein (LDL) receptor-related protein 1 (LRP1) [32], [33].…”

Anti-arthritic effect of astragaloside IV and its molecular mechanism

“…Increasing data show that IL-1-induced chondrocyte responses represent central pathogenic events in RA and osteoarthritis [26,28] . IL-1 in vitro directly inhibits cartilage proteoglycan synthesis and chondrocyte proliferation [29] .…”