MicroRNA-92a-3p Regulates Aggrecanase-1 and Aggrecanase-2 Expression in Chondrogenesis and IL-1β-Induced Catabolism in Human Articular Chondrocytes
MicroRNA-92a-3p Regulates Aggrecanase-1 and Aggrecanase-2 Expression in Chondrogenesis and IL-1β-Induced Catabolism in Human Articular Chondrocytes
复制标题
MicroRNA-92a-3p 调节软骨形成中聚集蛋白聚糖酶 1 和聚集蛋白聚糖酶 2 的表达以及人关节软骨细胞中 IL-1 β 诱导的分解代谢
DOI：
10.1159/000484579
复制
发表时间：
2017-01-01
影响因子：
--
通讯作者：
Kang, Yan
中科院分区：
文献类型：
--
作者：
Mao, Guping;Wu, Peihui;Kang, Yan
背景/目的：聚集蛋白聚糖酶-1 (ADAMTS-4) 和聚集蛋白聚糖酶-2 (ADAMTS-5) 是属于 ADAMTS（一种具有血小板反应蛋白基序的解整合素和金属蛋白酶）家族的分泌酶，在骨关节炎 (OA) 的进展中发挥重要作用。在这里，我们的目的是确定 ADAMTS-4/5 在软骨形成和炎症中的表达是否受到 microRNA-92a-3p (miR-92a-3p) 的调节。方法：通过定量聚合酶链式反应 (qPCR) 测定 MiR-92a-3p 和 ADAMTS-4/5 的表达。为了研究 miR-92a-3p 对 ADAMTS-4/5 表达的抑制作用，分别用成熟的 miR-92a-3p 或反义抑制剂 (anti-miR-92a-3p) 转染软骨形成人间充质干细胞 (hMSC) 和人软骨细胞。通过酶联免疫吸附测定 (ELISA) 定量 ADAMTS-4/5 蛋白的产生，并通过免疫印迹检查 miR-92a-3p 参与 IL-1 β 介导的分解代谢作用。通过使用特定抑制剂评估活化的 MAP 激酶 (MAPK) 和核因子 (NF)-κ B 的作用。通过荧光素酶报告基因测定证实了 miR-92a-3p 与其在 ADAMTS-4/5 mRNA 3'-非翻译区 (3'-UTR) 中的推定结合位点之间的相互作用。结果：miR-92a-3p 在成软骨 hMSC 中表达升高，而在 OA 软骨中的表达显着低于正常软骨。 IL-1 beta 刺激显着降低了原代人软骨细胞 (PHC) 中的 miR-92a-3p 表达。用 miR-92a-3p 转染软骨细胞可下调 IL-1 β 诱导的 ADAMTS-4/5 表达以及含有人 ADAMTS-4/5 mRNA 3'-UTR 的报告构建体的活性。 IL-1β 诱导软骨细胞中 MAPK 和 NF-κ B 的激活后，MiR-92a-3p 表达受到抑制。结论：MiR-92a-3p 是人软骨细胞中 ADAMTS-4/5 的重要调节因子，可能有助于 OA 的发生。 (c) 2017 年作者由 S. Karger AG 出版，巴塞尔
Background/Aims: Aggrecanase-1 (ADAMTS-4) and aggrecanase-2 (ADAMTS-5) are secreted enzymes belonging to the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family that play significant roles in the progression of osteoarthritis (OA). Here, we aimed to determine whether the expression of ADAMTS-4/5 in chondrogenesis and inflammation is regulated by microRNA-92a-3p (miR-92a-3p). Methods: MiR-92a-3p and ADAMTS-4/5 expressions were determined by quantitative polymerase chain reaction (qPCR). To investigate the repressive effect of miR-92a-3p on ADAMTS-4/5 expression, chondrogenic human mesenchymal stem cells (hMSCs) and human chondrocytes were transfected with mature miR-92a-3p or an antisense inhibitor (anti-miR-92a-3p), respectively. ADAMTS-4/5 protein production was quantified by enzyme-linked immunosorbent assay (ELISA), and miR-92a-3p involvement in IL-1 beta-mediated catabolic effects was examined by immunoblotting. The roles of activated MAP kinases (MAPK) and nuclear factor (NF)-kappa B were evaluated by using specific inhibitors. Interaction between miR-92a-3p and its putative binding site in the 3'-untranslated region (3'-UTR) of ADAMTS-4/5 mRNA was confirmed by luciferase reporter assay. Results: miR-92a-3p expression was elevated in chondrogenic hMSCs, with significantly lower expression in OA cartilage than in normal cartilage. Stimulation with IL-1 beta significantly reduced miR-92a-3p expression in primary human chondrocytes (PHCs). Transfection of chondrocytes with miR-92a-3p downregulated IL-1 beta-induced ADAMTS-4/5 expression, and the activity of a reporter construct containing the 3'-UTR of human ADAMTS-4/5 mRNA. MiR-92a-3p expression was suppressed upon IL-1 beta-induced activation of MAPK and NF-kappa B in chondrocytes. Conclusion: MiR-92a-3p is an important regulator of ADAMTS-4/5 in human chondrocytes and may contribute to the development of OA. (c) 2017 The Author(s) Published by S. Karger AG, Basel