LINC00707靶向miR-224-3p调控IL-1β诱导的软骨细胞损伤  

LINC00707 targets miR-224-3p to regulate IL-1β-induced chondrocyte damage

作　　者：徐显春[1] 齐保闯[1] 邱宇[1] XU Xianchun;QI Baochuang;QIU Yu(Department of Orthopedics,Yibin First People's Hospital,Yibin 644000,China)

机构地区：[1]宜宾市第一人民医院骨科,四川宜宾644000

出　　处：《现代医学》2021年第5期520-525,共6页Modern Medical Journal

摘　　要：目的:探讨长基因间非编码RNA00707(LINC00707)对白细胞介素(IL)-1β诱导的软骨细胞损伤的影响及其分子机制。方法:用10 ng·ml^(-1)的IL-1β处理人软骨细胞,记为IL-1β组,正常培养的细胞作为对照组,将LINC00707抑制表达载体质粒(si-LINC00707)及微小RNA-224-3p过表达载体(miR-224-3p)转染至软骨细胞后用10 ng·ml^(-1)的IL-1β处理,将LINC00707抑制表达载体质粒分别与miR-224-3p抑制表达载体共转染至软骨细胞后用10 ng·ml^(-1)的IL-1β处理;实时荧光定量PCR检测LINC00707和miR-224-3p的表达水平,酶联免疫吸附试验检测IL-6、干扰素γ(IFN-γ)水平,流式细胞术检测细胞凋亡,蛋白质印迹法检测蛋白表达,双荧光素酶报告实验验证LINC00707和miR-224-3p的靶向关系。结果:与对照组相比,IL-1β组LINC00707表达水平升高,miR-224-3p表达水平降低,IL-6、IFN-γ水平升高,细胞凋亡率升高,B细胞淋巴瘤/白血病-2(Bcl-2)表达水平降低,Bcl-2相关X(Bax)表达水平升高(P <0.05)。抑制LINC00707表达或过表达miR-224-3p后,IL-1β诱导的软骨细胞中IL-6、IFN-γ水平降低,细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低(P <0.05)。下调miR-224-3p表达逆转了抑制LINC00707表达对IL-1β诱导的软骨细胞损伤的保护作用。LINC00707靶向调控miR-224-3p的表达。结论:抑制LINC00707表达可能通过靶向上调miR-224-3p抑制IL-1β诱导的软骨细胞损伤。Objective: To explore the effects of long intergenic non-coding RNA00707( LINC00707) on interleukin-1β( IL-1β)-induced chondrocyte damage and its molecular mechanism. Methods: Human chondrocytes were treated with 10 ng·ml^(-1) IL-1β and recorded as IL-1β group;normally cultured cells were used as the control group;the LINC00707 suppression expression vector plasmid( si-LINC00707) and microRNA-224-3 p overexpression vector( miR-224-3 p) were transfected into chondrocytes and treated with 10 ng·ml^(-1) IL-1β;the LINC00707 suppression expression vector plasmid and miR-224-3 p suppression expression vector plasmid were cotransfected into chondrocytes and treated with 10 ng ·ml^(-1) IL-1β. Real-time fluorescent quantitative PCR( RTqPCR) was used to measure expression levels of LINC00707 and miR-224-3 p,enzyme-linked immunosorbent assay( ELISA) was used to measure interleukin 6( IL-6) and interferon gamma( IFN-gamma) levels,flow cytometry was used to detect cell apoptosis,Western blot was used to characterize protein expression,and the dual luciferase report experiment was used to verify the targeting relationship between LINC00707 and miR-224-3 p.Results: Compared with the control group,the LINC00707 expression level was higher in the IL-1β group,the miR-224-3 p expression level was lower,IL-6 and IFN-γ levels were lower,the cell apoptosis rate was higher,the expression level of B-cell lymphoma/leukemia-2( Bcl-2) was lower,and the expression level of Bcl-2 related X( Bax) was higher( P < 0. 05). After inhibiting the expression of LINC00707 or overexpressing miR-224-3 p,the levels of L-6 and IFN-γ in chondrocytes induced by IL-1β decreased,the apoptosis rate decreased,the expression of Bcl-2 increased,and the expression of Bax decreased( P < 0. 05). Down-regulating the expression of miR-224-3 p reverses the protective effect of inhibiting the expression of LINC00707 on IL-1β-induced chondrocyte damage.LINC00707 targets and regulates the expression of miR-224-3 p. Conclusion: Inhibiting the expression of L

关 键 词：长基因间非编码RNA00707 微小RNA-224-3p 软骨细胞 凋亡 炎症因子 损伤 

分 类 号：R684.3[医药卫生—骨科学]