Regulation of lncRNA-H19/miR-140-5p in cartilage matrix degradation and calcification in osteoarthritis.
Long non-coding RNA H19 (lncRNA H19)
calcification
cartilage
mechanism degradation
miR-140-5p
osteoarthritis (OA)
Journal
Annals of palliative medicine
ISSN: 2224-5839
Titre abrégé: Ann Palliat Med
Pays: China
ID NLM: 101585484
Informations de publication
Date de publication:
Jul 2020
Jul 2020
Historique:
received:
19
03
2020
accepted:
26
05
2020
pubmed:
25
6
2020
medline:
30
4
2021
entrez:
25
6
2020
Statut:
ppublish
Résumé
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. 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. 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 MMP1 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). 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.
Sections du résumé
BACKGROUND
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
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
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 MMP1 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
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.
Identifiants
pubmed: 32576007
pii: apm-20-929
doi: 10.21037/apm-20-929
doi:
Substances chimiques
MicroRNAs
0
Mirn140 microRNA, human
0
RNA, Long Noncoding
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM