ABSTRACT

Background

Osteoarthritis (OA) is a common degenerative joint disease in the elderly. Increasing evidence suggested that long non-coding RNAs (lncRNAs) played vital roles in OA progression. This study aimed to explore the role and mechanism of lncRNA small nucleolar RNA host gene 5 (SNHG5) in OA development.

Methods

Chondrocytes were stimulated with interleukin-1β (IL-1β) in vitro. The levels of SNHG5, miR-10a-5p, and H3 histone family 3B (H3F3B) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. Cell proliferation was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and colony formation assay. Cell apoptosis was tested by flow cytometry. The levels of apoptosis-related and cartilage-related markers were detected by western blot. The interaction among SNHG5, miR-10a-5p, and H3F3B was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.

Results

SNHG5 and H3F3B were downregulated, while miR-10a-5p was upregulated in OA cartilage tissues. Knockdown of SNHG5 enhanced IL-1β-induced apoptosis in chondrocytes. Rescue experiments verified that SNHG5 hindered apoptosis in IL-1β-stimulated chondrocytes by sponging miR-10a-5p. Moreover, H3F3B was a target of miR-10a-5p, and miR-10a-5p promoted IL-1β-induced chondrocyte apoptosis by regulating H3F3B. In addition, SNHG5 regulated H3F3B expression via sponging miR-10a-5p in IL-1β-treated chondrocytes.

Conclusion

SNHG5 suppressed chondrocytes apoptosis in OA by regulating the miR-10a-5p/H3F3B axis, which provided a promising biomarker for OA treatment.

中文翻译：

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LncRNA SNHG5通过调节miR-10a-5p/H3F3B轴促进骨关节炎软骨细胞增殖并抑制细胞凋亡

摘要

背景

骨关节炎（OA）是老年人常见的退行性关节病。越来越多的证据表明，长链非编码 RNA (lncRNA) 在 OA 进展中起着至关重要的作用。本研究旨在探讨lncRNA小核仁RNA宿主基因5（SNHG5）在OA发育中的作用和机制。

方法

在体外用白细胞介素-1β (IL-1β) 刺激软骨细胞。通过定量实时聚合酶链反应 (qRT-PCR) 或蛋白质印迹测量 SNHG5、miR-10a-5p 和 H3 组蛋白家族 3B (H3F3B) 的水平。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-溴化四唑 (MTT) 测定和集落形成测定来评估细胞增殖。通过流式细胞术检测细胞凋亡。通过蛋白质印迹检测细胞凋亡相关和软骨相关标志物的水平。SNHG5、miR-10a-5p 和 H3F3B 之间的相互作用通过双荧光素酶报告基因测定和 RNA 免疫沉淀 (RIP) 测定得到证实。

结果

SNHG5 和 H3F3B 下调，而 miR-10a-5p 在 OA 软骨组织中上调。敲除 SNHG5 可增强 IL-1β 诱导的软骨细胞凋亡。救援实验证实 SNHG5 通过海绵化 miR-10a-5p 阻碍 IL-1β 刺激的软骨细胞凋亡。此外，H3F3B 是 miR-10a-5p 的靶点，miR-10a-5p 通过调节 H3F3B 促进 IL-1β 诱导的软骨细胞凋亡。此外，SNHG5 通过海绵化 miR-10a-5p 在 IL-1β 处理的软骨细胞中调节 H3F3B 表达。

结论

SNHG5 通过调节 miR-10a-5p/H3F3B 轴来抑制 OA 中的软骨细胞凋亡，这为 OA 治疗提供了有希望的生物标志物。