Department of Spine Surgery and Orthopaedics, Xiangya Hospital Central South University, Changsha, China.
Phase I Clinical Trial Center, Xiangya Hospital Central South University, Changsha, China.
Anal Cell Pathol (Amst). 2023 Mar 29;2023:5911546. doi: 10.1155/2023/5911546. eCollection 2023.
The pathogenesis of osteoarthritis (OA) is complex and there is no specific drug for treatment. The aim of this study was to identify the molecular targets of OA therapy, focusing on the expression and biological functions of miR-182-5p and its target genes in OA.
miR-182-5p and fibroblast growth factor 9 (FGF9) were overexpressed or knocked down in IL-1-induced chondrocytes. An OA knee model was performed by surgically destroying the medial meniscus. The gene expression of miR-182-5p and FGF9 was calculated. The protein FGF9 was tested by western blotting. Cell counting kit-8 (CCK8), plate cloning assay, and flow cytometry were conducted to evaluate cell proliferation and apoptosis. The expression of inflammatory factors, tumor necrosis factor-alpha (TNF-), interleukin (IL)-6, and interleukin (IL)-8, was evaluated using enzyme-linked immunosorbent assay (ELISA). Dual-luciferase reporter assays validated the targeting relationship between miR-182-5p and FGF9. Hematoxylin-eosin (HE) and safranin O-fast Green (S-O) staining were utilized to access cartilage damage. Ki67 expression in cartilage was detected using immunohistochemistry (IHC). TdT-mediated dUTP nick-end labeling (TUNEL) assays were used to calculate the apoptosis rate of cartilage.
The expression of miR-182-5p was upregulated, and FGF9 was downregulated in the IL-1-induced chondrocytes. OA chondrocytes proliferation ability in the miR-182-5p mimics group was decreased, and the apoptosis rate and inflammatory factor were increased. Transfection with miR-182-5p inhibitor increased the proliferative ability and decreased the apoptosis rate in the IL-1-induced chondrocytes. Transfection with miR-182-5p inhibitor reversed IL-1-induced inflammatory factor release in chondrocytes. Targeted binding sites existed between miR-182-5p and FGF9. After overexpression of FGF9, the miR-182-5p effect on OA chondrocytes was reversed. The hyaline cartilage thickness and proteoglycan content decreased in OA rats, and this was reversed by miR-182-5p inhibitor treatment.
miR-182-5p expression levels were increased in OA chondrocytes and regulated chondrocyte proliferation, apoptosis, and inflammation by targeting FGF9. miR-182-5p is a potential gene for OA treatment.
骨关节炎（OA）的发病机制复杂，尚无特效药物治疗。本研究旨在确定 OA 治疗的分子靶点，重点研究 miR-182-5p 及其靶基因在 OA 中的表达和生物学功能。
在 IL-1 诱导的软骨细胞中过表达或敲低 miR-182-5p 和成纤维细胞生长因子 9（FGF9）。通过手术破坏内侧半月板建立 OA 膝关节模型。计算 miR-182-5p 和 FGF9 的基因表达。通过 Western blot 检测 FGF9 蛋白。通过细胞计数试剂盒-8（CCK8）、平板克隆试验和流式细胞术评估细胞增殖和凋亡。酶联免疫吸附试验（ELISA）评估炎症因子肿瘤坏死因子-α（TNF-）、白细胞介素（IL）-6 和白细胞介素（IL）-8 的表达。双荧光素酶报告实验验证 miR-182-5p 与 FGF9 的靶向关系。苏木精-伊红（HE）和番红 O-快绿（S-O）染色评估软骨损伤。免疫组化（IHC）检测软骨中 Ki67 的表达。TdT 介导的 dUTP 缺口末端标记（TUNEL）试验计算软骨细胞的凋亡率。
IL-1 诱导的软骨细胞中 miR-182-5p 表达上调，FGF9 表达下调。miR-182-5p 模拟物组 OA 软骨细胞增殖能力降低，凋亡率和炎症因子升高。转染 miR-182-5p 抑制剂增加了 IL-1 诱导的软骨细胞的增殖能力，降低了凋亡率。转染 miR-182-5p 抑制剂逆转了 IL-1 诱导的软骨细胞中炎症因子的释放。miR-182-5p 与 FGF9 之间存在靶向结合位点。过表达 FGF9 后，miR-182-5p 对 OA 软骨细胞的作用被逆转。OA 大鼠透明软骨厚度和糖胺聚糖含量降低，miR-182-5p 抑制剂治疗可逆转这一现象。
OA 软骨细胞中 miR-182-5p 表达水平升高，通过靶向 FGF9 调节软骨细胞增殖、凋亡和炎症。miR-182-5p 是 OA 治疗的潜在基因。
