Icariin improves chondrocyte function in osteoarthritis via the miR-29c/COL2A1 axis
10.13406/j.cnki.cyxb.003644
- VernacularTitle:淫羊藿苷通过miR-29c/COL2A1轴改善骨关节炎软骨细胞功能
- Author:
Zike HE
1
;
Shangzeng WANG
Author Information
1. 河南省中医院关节病科,郑州 450000
- Keywords:
osteoarthritis;
icariin;
microRNA-29cmiR-29c;
collagen type Ⅱ alpha 1 chain COL2A1;
cell apoptosis
- From:
Journal of Chongqing Medical University
2025;50(9):1271-1280
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of icariin(ICA)on chondrocyte function in osteoarthritis(OA)and its mechanism.Methods:After chondrocytes were treated with different concentrations of ICA(0.1,0.5,1.0,5.0,10.0 μmol/L),methylthiazolydiphenyl-tetrazolium(MTT)assay was used to measure the viability of chondrocytes,and then the optimal concentration of ICA was selected for subsequent experiments.Chondrocytes were stimulated by interleukin-1β(IL-1β,10 ng/mL)for 24 hours to establish a cell model of OA,and then the cells were treated with ICA at the optimal concentration of 1 μmol/L.Quantitative real-time polymerase chain reac-tion(RT-qPCR)was used to measure the expression level of mi-croRNA-29c(miR-29c);Western blot was used to measure the pro-tein expression level of collagen type Ⅱ alpha 1 chain(COL2A1);Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of matrix metalloproteinase-3(MMP-3)and tumor necro-sis factor-α(TNF-α)in the supernatant of cell culture;flow cytom-etry was used to measure cell apoptosis rate.Bioinformatics web-sites were used to predict the target genes of miR-29c,and dual-luciferase reporter assay was used to verify the targeted binding be-tween miR-29c and COL2A1.Functional rescue experiments were used to clarify the effect of ICA on inflammatory response and cell apoptosis in OA and its mechanism of action.Results:Compared with the control group,the IL-1β group had significant increases in the content of MMP-3(P<0.001)and TNF-α(P<0.001)in chondrocyte supernatant,a significant reduction in the expression level of COL2A1(P<0.001),and a significant increase in cell apoptosis rate(P<0.001).Compared with the IL-1β group,the ICA treatment group had significant reductions in the content of MMP-3(P=0.001;P<0.001)and TNF-α(P<0.001;P<0.001)in chondrocyte super-natant,a significant increase in the expression level of COL2A1(P<0.001;P<0.001),and a significant reduction in cell apoptosis rate(P<0.001;P<0.001).Compared with the control group,the IL-1β group had a significant increase in the expression level of miR-29c in chondrocytes(P<0.001),while ICA treatment significantly reduced the expression level of miR-29c(P<0.001).Compared with the control group,the miR-29c mimic group had significant increases in the content of MMP-3(P<0.001)and TNF-α(P<0.001)in chon-drocyte supernatant,a significant reduction in the expression level of COL2A1(P=0.001),and a significant increase in cell apoptosis rate(P<0.001);however,opposite results were obtained for the miR-29c inhibitor group.TargetScan7.1 database showed the exis-tence of binding sites between miR-29c and COL2A1.Dual-luciferase reporter assay showed that miR-29c mimic significantly inhib-ited the luciferase activity of wild-type COL2A1 vector(P<0.001),but with no influence on the luciferase activity of mutant COL2A1 vector(P=0.140).Knockdown of COL2A1 could partially reverse the effect of downregulated miR-29c on the secretion of MMP-3(P<0.001)and TNF-α(P<0.001),the protein expression level of COL2A1(P<0.001),and cell apoptosis rate(P<0.001)in chondro-cytes.ICA treatment could alleviate the influence of miR-29c overexpression on the secretion of MMP-3(P<0.001)and TNF-α(P<0.001),the protein expression level of COL2A1(P<0.001),and the apoptosis rate(P<0.001)of chondrocytes.Conclusion:ICA can alleviate the secretion of inflammatory cytokines and the apoptosis of chondrocytes induced by IL-1β,which may be associated with the miR-29c/COL2A1 axis..
