网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
LINC00707靶向miR-224-3p调控IL-1β诱导的软骨细胞损伤
作者:徐显春  齐保闯  邱宇 
单位:宜宾市第一人民医院 骨科, 四川 宜宾 644000
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
出版年·卷·期(页码):2021·49·第五期(520-525)
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-224-3p overexpression vector(miR-224-3p) were transfected into chondrocytes and treated with 10 ng·ml-1 IL-1β; the LINC00707 suppression expression vector plasmid and miR-224-3p suppression expression vector plasmid were co-transfected into chondrocytes and treated with 10 ng·ml-1 IL-1β. Real-time fluorescent quantitative PCR(RT-qPCR) was used to measure expression levels of LINC00707 and miR-224-3p, 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-224-3p. Results: Compared with the control group, the LINC00707 expression level was higher in the IL-1β group, the miR-224-3p 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-224-3p, 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-224-3p reverses the protective effect of inhibiting the expression of LINC00707 on IL-1β-induced chondrocyte damage. LINC00707 targets and regulates the expression of miR-224-3p. Conclusion: Inhibiting the expression of LINC00707 may inhibit IL-1β-induced chondrocyte damage by targeting the up-regulation of miR-224-3p.

参考文献:

[1] CHARLIER E,DEROYER C,CIREGIA F,et al.Chondrocyte dedifferentiation and osteoarthritis(OA)[J].Biochem Pharmacol,2019,165:49-65.
[2] 王新军,袁银鹏,王越,等.软骨细胞凋亡引发骨关节炎的机制研究进展[J].山东医药,2020,60(2):109-112.
[3] 张聘,张雷,赵建宁.LncRNA在软骨损伤与修复中作用机制的研究进展[J].医学研究生学报,2018,31(12):99-102.
[4] ZHU S,ZHOU Z,LI Z,et al.Suppression of LINC00707 alleviates lipopolysaccharide-induced inflammation and apoptosis in PC-12 cells by regulated miR-30a-5p/Neurod 1[J].Biosci Biotechnol Biochem,2019,83(11):2049-2056.
[5] LIU J,WU M,FENG G,et al.Downregulation of LINC00707 promotes osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by regulating DKK1 via targeting miR-103a-3p[J].Int J Mol Med,2020,46(3):1029-1038.
[6] 胡绍兰,孙蓓,韩菲,等.miR-224-3p通过抑制TLR4改善高糖诱导的大鼠肾小管上皮细胞的炎性反应[J].临床合理用药杂志,2017,10(22):52-53.
[7] DENG Y,MA G,DONG Q,et al.Overexpression of miR-224-3p alleviates apoptosis from cerebral ischemia reperfusion injury by targeting FIP200[J].J Cell Biochem,2019,120(10):17151-17158.
[8] 杨翔,林向进.骨性关节炎治疗方法的研究进展[J].中国骨与关节损伤杂志,2019,34(9):900-903.
[9] 郭鹏,范建楠,单霖,等.骨性关节炎软骨细胞凋亡的研究进展[J].中国社区医师,2019,35(6):8-11.
[10] 程环宇,李斯明,孟庆奇,等.骨性关节炎的发病机制与炎症反应的关系[J].中外医学研究,2020,18(6):185-188.
[11] 史达,赵聪喆,柴惠斌,等.IL-1β诱导内质网应激增加人软骨细胞凋亡[J].基础医学与临床,2015,35(8):1089-1093.
[12] 王西彬,左瑞庭.葫芦素B对IL-1β诱导的软骨细胞损伤及胞外基质代谢紊乱的影响[J].天津医药,2019,47(12):1210-1214.
[13] ZHAO Y,ZHAO J,GUO X,et al.Long non-coding RNA PVT1,a molecular sponge for miR-149,contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes[J].Biosci Rep,2018,38(5):BSR20180576.
[14] GAO G C,CHENG X G,WEI Q Q,et al.Long noncoding RNA MALAT-1 inhibits apoptosis and matrix metabolism disorder in interleukin-1β-induced inflammation in articular chondrocytes via the JNK signaling pathway[J].J Cell Biochem,2019,120(10):17167-17179.
[15] 洪军,刘景全,公方晓,等.微小RNA-224通过靶向抑制p21参与调控脂多糖诱导的肺微血管内皮细胞损伤研究[J].中华实验外科杂志,2020,37(1):81-83.
[16] LI H,LIU M W,YANG W,et al.Naringenin induces neuroprotection against homocysteine-induced PC12 cells via the upregulation of superoxide dismutase 1 expression by decreasing miR-224-3p expression[J].J Biol Regul Homeost Agents,2020,34(2):421-433.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 472206 位访问者


 ©《现代医学》编辑部
联系电话:025-83272481;83272479
电子邮件: xdyx@pub.seu.edu.cn

苏ICP备09058541