敲除miR-21-5p通过靶向颞下颌关节骨关节炎中的Gdf5减轻软骨基质降解。
Knockout of miR-21-5p alleviates cartilage matrix degradation by targeting Gdf5 in temporomandibular joint osteoarthritis.
作者信息
Zhang Aobo, Ma Shixing, Yuan Lingyu, Wu Shichao, Liu Shaopeng, Wei Xiang, Chen Lei, Ma Chuan, Zhao Huaqiang
机构信息
Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China.
The Institute for Tissue Engineering and Regenerative Medicine, Liaocheng People's Hospital, Liaocheng, China.
出版信息
Bone Joint Res. 2020 Oct;9(10):689-700. doi: 10.1302/2046-3758.910.BJR-2020-0140.R1.
AIMS
The study aimed to determine whether the microRNA miR21-5p (MiR21) mediates temporomandibular joint osteoarthritis (TMJ-OA) by targeting growth differentiation factor 5 (Gdf5).
METHODS
TMJ-OA was induced in MiR21 knockout (KO) mice and wild-type (WT) mice by a unilateral anterior crossbite (UAC) procedure. Mouse tissues exhibited histopathological changes, as assessed by: Safranin O, toluidine blue, and immunohistochemistry staining; western blotting (WB); and quantitative real-time polymerase chain reaction (RT-qPCR). Mouse condylar chondrocytes were transfected with a series of MiR21 mimic, MiR21 inhibitor, Gdf5 siRNA (si-GDF5), and flag-GDF5 constructs. The effects of MiR-21 and Gdf5 on the expression of OA related molecules were evaluated by immunofluorescence, alcian blue staining, WB, and RT-qPCR.
RESULTS
UAC altered the histological structure and extracellular matrix content of cartilage in the temporomandibular joint (TMJ), and KO of MiR21 alleviated this effect (p < 0.05). Upregulation of MiR21 influenced the expression of TMJ-OA related molecules in mandibular condylar chondrocytes via targeting Gdf5 (p < 0.05). Gdf5 overexpression significantly decreased matrix metalloproteinase 13 (MMP13) expression (p < 0.05) and reversed the effects of MiR21 (p < 0.05).
CONCLUSION
MiR21, which acts as a critical regulator of Gdf5 in chondrocytes, regulates TMJ-OA related molecules and is involved in cartilage matrix degradation, contributing to the progression of TMJ-OA. Cite this article: 2020;9(10):689-700.
目的
本研究旨在确定微小RNA miR21 - 5p（MiR21）是否通过靶向生长分化因子5（Gdf5）介导颞下颌关节骨关节炎（TMJ - OA）。
方法
通过单侧前牙反合（UAC）手术在MiR21基因敲除（KO）小鼠和野生型（WT）小鼠中诱导TMJ - OA。通过番红O、甲苯胺蓝和免疫组织化学染色、蛋白质免疫印迹法（WB）以及定量实时聚合酶链反应（RT - qPCR）评估小鼠组织的组织病理学变化。用一系列MiR21模拟物、MiR21抑制剂、Gdf5小干扰RNA（si - GDF5）和flag - GDF5构建体转染小鼠髁突软骨细胞。通过免疫荧光、阿尔辛蓝染色、WB和RT - qPCR评估MiR - 21和Gdf5对骨关节炎相关分子表达的影响。
结果
UAC改变了颞下颌关节（TMJ）软骨的组织结构和细胞外基质含量，MiR21基因敲除减轻了这种影响（p < 0.05）。MiR21的上调通过靶向Gdf5影响下颌髁突软骨细胞中TMJ - OA相关分子的表达（p < 0.05）。Gdf5过表达显著降低基质金属蛋白酶13（MMP13）的表达（p < 0.05）并逆转了MiR21的作用（p < 0.05）。
结论
MiR21作为软骨细胞中Gdf5的关键调节因子，调节TMJ - OA相关分子并参与软骨基质降解，促进TMJ - OA的进展。引用本文：2020；9(10):689 - 700。
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