长链非编码核糖核酸长基因非编码核糖核酸-p53诱导转录本对膝骨关节炎软骨细胞增殖、凋亡及炎性因子分泌的影响及其机制  

Effect of long non-coding ribonucleic acid long intergenic non-protein coding ribonucleic acid p53-induced transcript targeting microRNA-628-5p gene on the proliferation,apoptosis and secretion of inflammatory factors in knee osteoarthritis chondrocytes

作　　者：吴迪[1] 司丽娜[2] 罗金伟 吕永明[1] 杨松鹤[2] Wu Di;Si Lina;Luo Jinwei;Lyu Yongming;Yang Songhe(Department of Joint Surgery,Affiliated Hospital of Chengde Medical College,Chengde 067000,China;Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院附属医院关节外科,067000 [2]承德医学院,067000

出　　处：《中华实验外科杂志》2022年第2期325-328,共4页Chinese Journal of Experimental Surgery

基　　金：承德市科学技术研究与发展计划项目(202109A034)。

摘　　要：目的探讨长链非编码核糖核酸长基因非编码核糖核酸-p53诱导转录本(lncRNA LINC-PINT)对膝骨关节炎软骨细胞增殖、凋亡及炎性因子分泌的影响及其机制。方法根据处理方法将人膝关节软骨细胞随机分为对照组(NC组)、白细胞介素(IL)-1β处理组(IL-1β组)、IL-1β+pcDNA-lncRNA LINC-PINT组、IL-1β+pcDNA组、IL-1β+anti-微小核糖核酸(miR)-628-5p组、IL-1β+anti-miR-NC组、IL-1β+pcDNA-lncRNA LINC-PINT+miR-NC组和IL-1β+pcDNA-lncRNA LINC-PINT+miR-628-5p组。实时荧光定量聚合酶链反应(RT-qPCR)检测lncRNA LINC-PINT和miR-628-5p表达;流式细胞仪检测细胞凋亡率;蛋白质印迹法检测B细胞淋巴瘤-2相关X蛋白(bax)和裂解的半胱氨酰天冬氨酸特异性蛋白酶-3(cleaved Caspase-3)蛋白表达;酶联免疫吸附法检测肿瘤坏死因子-α(TNF-α)和IL-6水平;双荧光素酶报告实验检测lncRNA LINC-PINT和miR-628-5p的靶向关系。两组间比较行t检验, 多组间比较采用单因素方差分析, 组间两两比较采用LSD-t检验。结果 IL-1β组lncRNA LINC-PINT表达低于NC组(0.35±0.03比1.00±0.11), miR-628-5p表达高于NC组(2.13±0.15比0.97±0.06), 差异有统计学意义(t=17.103、18.804, P<0.05)。IL-1β+pcDNA-lncRNA LINC-PINT组细胞活性高于IL-1β+pcDNA(1.08±0.07比0.57±0.04), bax(0.41±0.04比0.82±0.07)、cleaved Caspase-3(0.46±0.03比0.91±0.06)、细胞凋亡率[(9.72±0.72)%比(25.38±2.31)%]、TNF-α[(93.51±8.36) pg/ml比(218.76±18.03) pg/ml]和IL-6[(102.18±8.72) pg/ml比(263.91±22.56) pg/ml]水平低于IL-1β+pcDNA组, 差异有统计学意义(t=29.122、15.356、10.422、11.753、6.755、16.023, P<0.05)。IL-1β+pcDNA-lncRNA LINC-PINT+miR-628-5p组miR-628-5p(1.95±0.16比1.00±0.10)、bax(0.91±0.06比0.42±0.03)、cleaved Caspase-3(0.82±0.08比0.46±0.05)、细胞凋亡率[(26.38±2.15)%比(9.61±0.78)%]、TNF-α[(197.53±18.35) pg/ml比(91.35±8.26) pg/ml]和IL-6水平[(228.67±19.84) pg/ml比(101.54±8.39) pg/ml]高于IL-1β+pcDNA-lncRNA LINC-PINT+miR-NCObjective To study the effect of long non-coding ribonucleic acid long intergenic non-protein coding ribonucleic acid p53-induced transcript(lncRNA LINC-PINT)on the proliferation,apoptosis and secretion of inflammatory factors in knee osteoarthritis chondrocytes and its mechanism.Methods Chondrocytes were randomly divided into NC group,interleukin(IL)-1βgroup,IL-1β+pcDNA-lncRNA LNC-PINT group,IL-1β+pcDNA group,IL-1β+anti-micro ribonucleic acid(miR)-628-5p group,IL-1β+anti-miR-NC group,IL-1β+pcDNA-lncRNA LINC-PINT+miR-NC group,IL-1β+pcDNA-lncRNA LINC-PINT+miR-628-5p group.The real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of lncRNA LINC-PINT and miR-628-5p.The flow cytometry was used to detect apoptosis rate.Western blotting was used to detect protein expression of B-cell lymphoma 2 associated X Protein(bax)and cleaved cysteinyl aspartate-specific protease-3(cleaved Caspase-3).The enzyme-linked immunosorbent assay was used to detect the levels of tumor necrosis factor-α(TNF-α)and IL-6.The dual luciferase reporter experiment was done to detect the targeting relationship between lncRNA Lnc-PINT and miR-628-5p.The t test was used for comparison between two groups,the one-way analysis of variance was used for comparison between multiple groups,and the LSD-t test was used for pairwise comparison between groups.Results The expression of lncRNA LINC-PINT was lower(2.13±0.15 vs.0.97±0.06)and that of miR-628-5p was higher(0.35±0.03 vs.1.00±0.11)in IL-1βgroup than that in NC group with the differences being statistically significant(t=17.103,18.804,P<0.05).The cell viability of IL-1β+pcDNA-lncRNA LINC-PINT group was higher than that of IL-1β+pcDNA group(1.08±0.07 vs.0.57±0.04),and the expression of bax(0.41±0.04 vs.0.82±0.07)and cleaved Caspase-3(0.46±0.03 vs.0.91±0.06),apoptosis rate[(9.72±0.72)%vs.(25.38±2.31)%],TNF-α[(93.51±8.36)vs.(218.76±18.03)pg/ml]and IL-6[(102.18±8.72)vs.(263.91±22.56)pg/ml]were lower than in IL-1β+pcDNA gro

关 键 词：膝骨关节炎 微小RNA 炎性因子 

分 类 号：R684.3[医药卫生—骨科学]