LncRNA ZBED3-AS1靶向miR-506-5p对骨关节炎软骨细胞增殖、分化的影响OA
LncRNA ZBED3-AS1 Regulates Proliferation and Differentiation of Osteoarthritic Chondrocytes by Targeting miR-506-5p
目的 分析长链非编码RNA ZBED3反义RNA1(LncRNA ZBED3-AS1)靶向微小RNA(miR)-506-5p对骨关节炎(OA)软骨细胞增殖、分化的影响,为开发更有效的治疗策略提供理论依据.方法 正常培养人软骨细胞CHON-001为Control组,采用白细胞介素-1β(IL-1β)诱导人软骨细胞CHON-001建立OA 软骨细胞模型,并随机分为 Model 组、OE-NC 组、OE-LncRNA ZBED3-AS1 组、OE-LncRNA ZBED3-AS1+pcDNA-NC 组、OE-LncRNA ZBED3-AS1+pcDNA-miR-506-5p 组.采用实时荧光定量 PCR 检测 LncRNA ZBED3-AS1、miR-506-5p 表达;双荧光素酶报告检测 LncRNA ZBED3-AS1 和 miR-506-5p的靶向关系;CCK8检测细胞增殖;流式细胞仪检测细胞凋亡;实时荧光定量PCR和Western blot法检测增殖细胞核抗原(PCNA)、骨形成蛋白2(BMP-2)和Runt相关转录因子2(Runx2)的mRNA和蛋白表达水平.结果 与 Control 组相比,Model 组细胞中 LncRNA ZBED3-AS1、存活率及 PCNA、BMP-2、Runx2 mRNA 和蛋白表达下降,而miR-506-5p、细胞凋亡率表达上升(P＜0.05);与Model组或OE-NC组相比,OE-Ln-cRNA ZBED3-AS1 组细胞中 LncRNA ZBED3-AS1、存活率及 PCNA、BMP-2、Runx2 mRNA 和蛋白表达上升,而 miR-506-5p、细胞凋亡率表达下降(P＜0.05);与 OE-LncRNA ZBED3-AS1 组或 OE-LncRNA ZBED3-AS1+pcDNA-NC 组相比,OE-LncRNA ZBED3-AS1+pcDNA-miR-506-5p 组细胞中存活率及PCNA、BMP-2、Runx2mRNA和蛋白表达下降,miR-506-5p、细胞凋亡率表达上升(P＜0.05).双荧光素酶报告结果显示,WT-LncRNA ZBED3-AS1+miR-506-3p mimic 组与 WT-LncRNA ZBED3-AS1+mimic-NC组相比,细胞荧光素酶活性降低(P＜0.05).结论 LncRNA ZBED3-AS1通过靶向抑制miR-506-5p调控OA软骨细胞的增殖、分化.
Objective To investigate the effects of long non-coding RNA ZBED3 antisense RNA 1(lncRNA ZBED3-AS1)targeting microRNA-506-5p(miR-506-5p)on the proliferation and differentiation of osteoarthritic(OA)chondrocytes,and to provide a theoretical basis for the development of novel therapeutic strategies for osteoarthritis.Methods Human chondrocytes(CHON-001)cultured under normal conditions were used as the control group.An OA chondrocyte model was established by interleukin-1β(IL-1β)stimulation and randomly divided into the following groups:Model,OE-NC,OE-lncRNA ZBED3-AS1,OE-lncRNA ZBED3-AS1+pcDNA-NC,and OE-lncRNA ZBED3-AS1+pcDNA-miR-506-5p.The expression levels of lncRNA ZBED3-AS1 and miR-506-5p were de-termined by quantitative real-time PCR(qRT-PCR).The targeting relationship between lncRNA ZBED3-AS1 and miR-506-5p was verified using a dual-luciferase reporter assay.Cell proliferation was assessed by CCK-8 assay,and apoptosis was analyzed by flow cytometry.The mRNA and protein expression levels of proliferating cell nuclear antigen(PCNA),bone morphogenetic protein 2(BMP-2),and Runt-related transcription factor 2(Runx2)were detected by qRT-PCR and Western blotting,respectively.Results Compared with the control group,the model group showed sig-nificantly decreased expression of lncRNA ZBED3-AS1,reduced cell viability,and downregulated mRNA and protein levels of PCNA,BMP-2,and Runx2,while miR-506-5p expression and apoptosis rate were significantly increased(P＜0.05).Compared with the model or OE-NC groups,overexpression of lncRNA ZBED3-AS1 significantly in-creased cell viability and the expression of PCNA,BMP-2,and Runx2,while decreasing miR-506-5p expression and apoptosis rate(P＜0.05).These effects were partially reversed by co-transfection with pcDNA-miR-506-5p.Dual-luciferase reporter assays demonstrated that miR-506-5p significantly reduced luciferase activity in cells transfected with wild-type lncRNA ZBED3-AS1 constructs(P＜0.05),confirming a direct targeting interaction.Conclusion LncRNA ZBED3-AS1 promotes proliferation and differentiation and inhibits apoptosis of osteoarthritic chondrocytes by targeting and suppressing miR-506-5p.This regulatory axis may represent a potential therapeutic target for osteoarthri-tis.
李炳南;李奎蒙;刘鹏飞
张家口市第一医院骨三科(河北 张家口 075000)张家口市第一医院骨三科(河北 张家口 075000)张家口市第一医院骨三科(河北 张家口 075000)
医药卫生
长链非编码RNA ZBED3反义RNA1微小RNA-506-5p骨关节炎软骨细胞增殖软骨细胞分化
LncRNA ZBED3-AS1miR-506-5posteoarthritischondrocyte proliferationchondrocyte dif-ferentiation
《广东医学》 2026 (2)
194-200,7
张家口市重点研发计划项目(2021023D)
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