Regulation of lncRNA-H19/miR-140-5p in cartilage matrix degradation and calcification in osteoarthritis

Abstract

Background: To study the effect of long non-coding RNA H19/microRNA (miR)-140-5p regulatory axis on the degradation and calcification of osteoarthritis (OA) cartilage matrix.Methods: Twenty cases of clinical OA cartilage tissue and healthy cartilage tissue were collected.Construction of small interfering RNA vector (si-H19) and negative control (si-Control), miR-140-5p mimic and negative control mimic-NC, miR-140-5p inhibitor, and negative control inhibitor-NC transfected human chondrocytes HC-A cells. Chondrocytes were categorized into classes of si-H19, si-Control, mimic, mimic-NC, inhibitor, NC inhibitor, inhibitor + si-H19, NC inhibitor + si-H19. The expression levels of H19, miR-140-5p, alkaline phosphatase (ALP), osteocalcin (OCN), and bone sialoprotein (BSP) in cartilage tissue and chondrocytes were detected using real-time fluorescent quantitative PCR (RT-qPCR). In each group of chondrocytes, Western blotting was used to detect the levels of matrix metalloproteinase (MMP)-1, MMP-13, and type II α1 collagen fibers (COL2A1). CCK-8 was used to detect the cell proliferation rate, and flow cytometry was used to detect the cell apoptosis rate.Results: H19 was up-regulated in OA cartilage tissue (P<0.05); Silencing H19 not only prevented chondrocyte apoptosis but also facilitated chondrocyte proliferation (both P<0.05), down-regulated MMP-1 and MMP-13 expression levels (both P<0.05), and up-regulated COL2A1 expression rates (P<0.05). Also, silenced H19 inhibited the mRNA level of ALP, OCN, and BSP and ALP activity of chondrocytes (all P<0.05). There was a negative correlation between miR-140-5p and H19 in OA cartilage tissue and chondrocytes (r=-0.98, P<0.001). Silenced miR-140-5p significantly reversed the inhibitory effects of silenced H19 on matrix degradation and calcification markers in chondrocytes (all P<0.05). Conclusions:The present study showed that the H19/ miR-140-5p regulatory axis could affect the degradation and calcification of the cartilage matrix in OA cells.

The umbilical cord mesenchymal stem cell‐derived exosomal lncRNA H19 improves osteochondral activity through miR‐29b‐3p/FoxO3 axis

“…It is possible that the H19/miR-106a-5p/TCF4 axis may be responsible for the progression of OA. Yang et al 27 confirmed that silencing lncRNA H19 prevented chondrocyte apoptosis, promoted proliferation, and downregulated MMP13 levels.…”

Platelet-rich plasma combined with ozone prevents cartilage destruction and improves weight-bearing asymmetry in a surgery-induced osteoarthritis rabbit model

“…It is reported that the MMP family plays a primary role in progressive OA, through a critical role in the degradative regulation of cartilage matrix components (36). MMP-1 is a rate-limiting type II collagenase causing degradation and destruction of extracellular matrix and activates other MMP family members to play an essential role in OA (37). Consistent with the results of previous studies, the Hulth-induced OA rabbits showed elevated expression of BMP-2 mRNA in joint fluid, and MMP-1 expression in the articular cartilage.…”

MicroRNA-140-5p represses chondrocyte pyroptosis and relieves cartilage injury in osteoarthritis by inhibiting cathepsin B/Nod-like receptor protein 3

“…In line with our findings, a prior study has revealed that miR-140-5p is essential for in vitro chondrogenesis whilst being down-regulated in osteoarthritic cartilages [ 30 ]. Moreover, miR-140-5p is indicated to protect chondrocytes from inflammatory injury during the pathogenesis of OA [ 31 ], whereas another prior investigation has uncovered that inhibition of miR-140-5p promotes the cartilage matrix degradation in OA cells [ 32 ]. Altogether, these findings and evidence indicate that OA induction inhibits miR-140-5p expression, while over-expression of miR-140-5p alleviates cartilage degradation and inflammation.…”