MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes
MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes
复制标题
DOI：
10.3390/cells8080874
复制
发表时间：
2019-08-01
期刊：
影响因子：
6
通讯作者：
Fioravanti, Antonella
中科院分区：
文献类型：
--
作者：
Cheleschi, Sara;Tenti, Sara;Fioravanti, Antonella
目前的证据表明，在骨关节炎(OA)的发病机制中，脂肪因子和microRNA(MiRNA)之间存在复杂的相互作用。本研究探讨miR-34a和miR-181a在调节内脂素诱导的人骨关节炎软骨细胞凋亡和氧化应激中的作用。将miR-34a和miR-181a抑制剂导入软骨细胞，加入核因子-kappaB抑制剂(Bay-11-7082，预孵育2 h)，用Visfatin刺激软骨细胞24 h。用实时定量聚合酶链式反应(Real Time PCR)和免疫印迹法检测细胞凋亡和活性氧(ROS)的产生，用实时定量聚合酶链式反应(Real Time PCR)和免疫印迹法检测抗氧化酶、核因子红系(NRF)2和B细胞淋巴瘤(BCL)2的表达。免疫荧光法检测P50核因子-kappaB亚单位。Visfatin可显著诱导骨性关节炎软骨细胞的凋亡和超氧阴离子产生，增加miR-34a、miR-181a、超氧化物歧化酶(SOD)-2、过氧化氢酶(CAT)、NRF2的表达，降低bcl2基因和蛋白的表达。用miR-34a和miR-181a抑制剂可抑制内脂素引起的所有效应。与Bay-11-7082预孵育可拮抗Visfatin诱导的miRNA、bcl2、SOD-2、CAT和NRF2的表达。抑制miR-34a和miR-181a可显著降低p50 NF-kappa B的活性，Visfatin证实了其诱导人骨关节炎软骨细胞凋亡和氧化应激的能力；这些作用可能是由miR-34a和miR-181a通过NF-kappa B途径介导的。我们强调内脂素作为治疗骨性关节炎的潜在靶点的相关性。
Current evidence suggests a complex interaction between adipokines and microRNA (miRNA) in osteoarthritis (OA) pathogenesis. The present study explored the role of miR-34a and miR-181a in regulating apoptosis and oxidative stress induced by visfatin in human OA chondrocytes. Chondrocytes were transfected with miR-34a and miR-181a inhibitors and stimulated with visfatin for 24 h, in the presence of nuclear factor (NF)-kappa B inhibitor (BAY-11-7082, 2 h pre-incubation). Apoptosis and reactive oxygen species (ROS) production were detected by cytometry, miRNA, antioxidant enzymes, nuclear factor erythroid (NRF)2 and B-cell lymphoma (BCL)2 expressions by quantitative real time polymerase chain reaction (real time PCR) and western blot. P50 NF-kappa B subunit was measured by immunofluorescence. Visfatin significantly induced apoptosis and superoxide anion production, increased miR-34a, miR-181a, superoxide dismutase (SOD)-2, catalase (CAT), NRF2 and decreased BCL2 gene and protein expression in OA chondrocytes. All the visfatin-caused effects were suppressed by using miR-34a and miR-181a inhibitors. Pre-incubation with BAY-11-7082 counteracted visfatin-induced expression of miRNA, BCL2, SOD-2, CAT and NRF2. Inhibition of miR-34a and miR-181a significantly reduced the activation of p50 NF-kappa B. Visfatin confirms its ability to induce apoptosis and oxidative stress in human OA chondrocytes; these effects appeared mediated by miR-34a and miR-181a via NF-kappa B pathway. We highlight the relevance of visfatin as potential therapeutic target for OA treatment.