hsa_circ_0005105靶向miR-508-3p对关节炎软骨细胞的影响
Effects of hsa_circ_0005105 targeting miR-508-3p on arthritic chondrocytes
  
DOI:10.3969/j.issn.1006.7108.2022.10.005
中文关键词:  hsa_circ_0005105  miR-508-3p  关节炎  软骨细胞  增殖
英文关键词:hsa_circ_0005105  miR-508-3p  arthritis  chondrocytes  proliferation
基金项目:2021年武汉市医学科研项目(编号WX21A01)
作者单位
张明焕 毛文* 湖北省武汉市第三医院骨科湖北 武汉 430071 
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中文摘要:
      目的 探讨hsa_circ_0005105对关节炎软骨细胞增殖、凋亡及炎症因子分泌的影响及机制。方法 实时荧光定量PCR(RT-qPCR)检测软骨细胞中hsa_circ_0005105和miR-508-3p的表达水平;将软骨细胞分为NC组、IL-1β组、IL-1β+si-NC组、IL-1β+si-hsa_circ_0005105组、IL-1β+miR-NC组、IL-1β+miR-508-3p组、IL-1β+si-hsa_circ_0005105+anti-miR-NC组、IL-1β+si-hsa_circ_0005105+anti-miR-508-3p组;蛋白质印迹法检测凋亡蛋白表达;流式细胞术检测凋亡;CCK-8与平板克隆形成能力检测细胞增殖能力;酶联免疫吸附法检测TNF-α、IL-6水平;双荧光素酶报告实验检测hsa_circ_0005105和miR-508-3p的靶向关系。结果 IL-1β诱导的软骨细胞中hsa_circ_0005105表达水平升高,miR-508-3p表达水平降低(P<0.05)。低表达hsa_circ_0005105或过表达miR-508-3p可降低IL-1β诱导的软骨细胞Bax、Cleaved-caspase-9、Cleaved-caspase-3相对表达水平、细胞凋亡率,升高Bcl-2蛋白水平、细胞活性,细胞克隆形成数增多,TNF-α和IL-6水平降低(P<0.05)。hsa_circ_0005105靶向调控miR-508-3p;低表达miR-508-3p可以逆转hsa_circ_0005105低表达对IL-1β诱导的软骨细胞损伤及炎症因子分泌的影响。结论 低表达hsa_circ_0005105通过靶向上调miR-508-3p抑制IL-1β诱导的软骨细胞凋亡及炎症因子分泌,促进细胞增殖。
英文摘要:
      Objective To explore the effect and mechanism of hsa_circ_0005105 on the proliferation, apoptosis and secretion of inflammatory factors of arthritis chondrocytes. Methods Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression levels of hsa_circ_0005105 and miR-508-3p in chondrocytes; chondrocytes were divided into NC group, IL-1β group, IL-1β+si-NC group, IL-1β+si-hsa_circ_0005105 group, IL-1β+miR-NC group, IL-1β+miR-508-3p group, IL-1β+si-hsa_circ_0005105+anti-miR-NC group, IL-1β+si-hsa_circ_ 0005105+anti-miR-508-3p group; apoptosis protein expression was detected by Western blotting; apoptosis was detected by flow cytometry; CCK-8 and plate clone formation ability were used to detect cell proliferation ability; enzyme-linked immunosorbent assay to detect levels of TNF-α and IL-6; dual luciferase reporter experiment to detect the targeting relationship between hsa_circ_0005105 and miR-508-3p. Results The expression level of hsa_circ_0005105 in IL-1β-induced chondrocytes was increased, and the expression level of miR-508-3p was decreased (P<0.05). Low expression of hsa_circ_0005105 or overexpression of miR-508-3p could reduce the relative expression levels of Bax, Cleaved-caspase-9 and Cleaved-caspase-3, and the apoptosis rate of chondrocytes induced by IL-1β, and increase the level of Bcl-2 protein, Cell viability, the number of cell clones increased, and the levels of TNF-α and IL-6 decreased (P<0.05). hsa_circ_0005105 targets and regulates miR-508-3p; low expression of miR-508-3p could reverse the effect of low expression of hsa_circ_0005105 on IL-1β-induced chondrocyte damage and inflammatory factor secretion. Conclusions The low expression of hsa_circ_0005105 can inhibit IL-1β-induced chondrocyte apoptosis and secretion of inflammatory factors by targeting up-regulation of miR-508-3p, and promote cell proliferation.
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