抑制miR-182-5p靶向FGF9缓解骨关节炎。

IF 2.6 4区 医学 Q3 CELL BIOLOGY
Yang Sun, Sanmao Su, Mengjun Li, Ang Deng
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引用次数: 3

摘要

背景:骨关节炎(OA)的发病机制复杂,目前尚无特征性的治疗药物。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。方法:miR-182-5p和成纤维细胞生长因子9 (FGF9)在il -1β诱导的软骨细胞中过表达或下调。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的基因表达量。western blotting检测FGF9蛋白表达。细胞计数试剂盒-8 (CCK8)、平板克隆实验和流式细胞术检测细胞增殖和凋亡情况。采用酶联免疫吸附试验(ELISA)评估炎症因子、肿瘤坏死因子-α (TNF-α)、白细胞介素(IL)-6和白细胞介素(IL)-8的表达。双荧光素酶报告基因试验验证了miR-182-5p和FGF9之间的靶向关系。采用苏木精-伊红(HE)和红花素O-fast Green (S-O)染色观察软骨损伤。免疫组化(IHC)检测软骨组织中Ki67的表达。采用tdt介导的dUTP镍端标记法(TUNEL)计算软骨细胞凋亡率。结果:il -1β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软骨细胞增殖能力下降,凋亡率和炎症因子升高。转染miR-182-5p抑制剂可提高il -1β诱导的软骨细胞的增殖能力,降低其凋亡率。转染miR-182-5p抑制剂可逆转il -1β诱导的软骨细胞炎症因子释放。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蛋白多糖含量下降,miR-182-5p抑制剂治疗可逆转这种情况。结论:miR-182-5p在OA软骨细胞中的表达水平升高,并通过靶向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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Inhibition of miR-182-5p Targets FGF9 to Alleviate Osteoarthritis.

Background: 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.

Methods: miR-182-5p and fibroblast growth factor 9 (FGF9) were overexpressed or knocked down in IL-1β-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-α), 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.

Results: The expression of miR-182-5p was upregulated, and FGF9 was downregulated in the IL-1β-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-1β-induced chondrocytes. Transfection with miR-182-5p inhibitor reversed IL-1β-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.

Conclusions: 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.

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来源期刊
Analytical Cellular Pathology
Analytical Cellular Pathology ONCOLOGY-CELL BIOLOGY
CiteScore
4.90
自引率
3.10%
发文量
70
审稿时长
16 weeks
期刊介绍: Analytical Cellular Pathology is a peer-reviewed, Open Access journal that provides a forum for scientists, medical practitioners and pathologists working in the area of cellular pathology. The journal publishes original research articles, review articles, and clinical studies related to cytology, carcinogenesis, cell receptors, biomarkers, diagnostic pathology, immunopathology, and hematology.
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