Jin Y, Chen X, Gao Z Y, Liu K, Hou Y, Zheng J
Department of Orthopaedics, Henan Provincial People's Hospital (Zhengzhou University People's Hospital), Zhengzhou 450003, China.
Department of Orthopaedics, Henan Provincial People's Hospital (Zhengzhou University People's Hospital), Zhengzhou 450003, China
Bone Joint Res. 2017 Apr;6(4):196-203. doi: 10.1302/2046-3758.64.BJR-2016-0224.R1.
This study aimed to explore the role of miR-320a in the pathogenesis of osteoarthritis (OA).
Human cartilage cells (C28/I2) were transfected with miR-320a or antisense oligonucleotides (ASO)-miR-320a, and treated with IL-1β. Subsequently the expression of collagen type II alpha 1 (Col2α1) and aggrecan (ACAN), and the concentrations of sulfated glycosaminoglycans (sGAG) and matrix metallopeptidase 13 (MMP-13), were assessed. Luciferase reporter assay, qRT-PCR, and Western blot were performed to explore whether pre-B-cell leukemia Homeobox 3 (PBX3) was a target of miR-320a. Furthermore, cells were co-transfected with miR-320a and PBX3 expressing vector, or cells were transfected with miR-320a and treated with a nuclear factor kappa B (NF-κB) antagonist MG132. The changes in Col2α1 and ACAN expression, and in sGAG and MMP-13 concentrations, were measured again. Statistical comparisons were made between two groups by using the two-tailed paired -test.
Expression of miR-320a was elevated in OA cartilage tissues and chondrocytes, and in IL-1β-stimulated C28/I2 cells (p < 0.05 or p < 0.01). MiR-320a overexpression enhanced IL-1β-induced down-regulation of Col2α1 and ACAN and sGAG, and increased the IL-1β-induced overexpression of MMP-13 (p < 0.01). PBX3 was a direct target of miR-320a. PBX3 and MG132 co-transfection attenuated the effects of miR-320a on the expression of Col2α1, ACAN, sGAG and MMP-13(p < 0.01).
Overexpression of miR-320a might enhance IL-1β-induced cartilage degradation factors. These effects might be via targeting PBX3 and regulating NF-κB. Y. Jin, X. Chen, Z. Y. Gao, K. Liu, Y. Hou, J. Zheng. The role of miR-320a and IL-1β in human chondrocyte degradation. 2017;6:-203. DOI: 10.1302/2046-3758.64.BJR-2016-0224.R1.
本研究旨在探讨miR - 320a在骨关节炎（OA）发病机制中的作用。
将miR - 320a或反义寡核苷酸（ASO）- miR - 320a转染人软骨细胞（C28/I2），并用白细胞介素 - 1β（IL - 1β）处理。随后评估Ⅱ型胶原蛋白α1（Col2α1）和聚集蛋白聚糖（ACAN）的表达，以及硫酸化糖胺聚糖（sGAG）和基质金属蛋白酶13（MMP - 13）的浓度。进行荧光素酶报告基因检测、qRT - PCR和蛋白质免疫印迹法以探究前B细胞白血病同源盒3（PBX3）是否为miR - 320a的靶标。此外，将细胞与miR - 320a和PBX3表达载体共转染，或者将细胞用miR - 320a转染并使用核因子κB（NF - κB）拮抗剂MG132处理。再次测量Col2α1和ACAN表达以及sGAG和MMP - 13浓度的变化。使用双侧配对t检验在两组之间进行统计学比较。
miR - 320a在OA软骨组织和软骨细胞以及IL - 1β刺激的C28/I2细胞中表达升高（p < 0.05或p < 0.01）。miR - 320a过表达增强了IL - 1β诱导的Col2α1、ACAN和sGAG的下调，并增加了IL - 1β诱导的MMP - 13过表达（p < 0.01）。PBX3是miR - 320a的直接靶标。PBX3与MG132共转染减弱了miR - 320a对Col2α1、ACAN、sGAG和MMP - 13表达的影响（p < 0.01）。
miR - 320a的过表达可能增强IL - 1β诱导的软骨降解因子。这些作用可能是通过靶向PBX3并调节NF - κB实现的。Y. Jin，X. Chen，Z.Y. Gao，K. Liu，Y. Hou，J. Zheng。miR - 320a和IL - 1β在人软骨细胞降解中的作用。2017；6：- 203。DOI：10.1302/2046 - 3758.64.BJR - 2016 - 0224.R1。
