[1]符传恭),陆志夫),宋世锋),等.miR-424-5p靶向PHD2对骨性关节炎软骨细胞凋亡及炎症因子分泌的影响[J].郑州大学学报(医学版),2021,(01):1-5.
　FU Chuangong),LU Zhifu),SONG Shifeng),et al.Effects of miR-424-5p targeting PHD2 on apoptosis and inflammatory factor secretion in osteoarthritis chondrocytes[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2021,(01):1-5.
点击复制

miR-424-5p靶向PHD2对骨性关节炎软骨细胞凋亡及炎症因子分泌的影响()
分享到：

《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2021年01期
页码:
1-5
栏目:
论著
出版日期:
2021-01-20

文章信息/Info

Title:
Effects of miR-424-5p targeting PHD2 on apoptosis and inflammatory factor secretion in osteoarthritis chondrocytes
作者:
符传恭1);  陆志夫1);  宋世锋2); 黄 健1); 苟学健1)
1)海口市中医医院骨科 海口 570000 2)海南医学院第二附属医院骨二科 海口 570000
Author(s):
FU Chuangong1);  LU Zhifu1);  SONG Shifeng2);  HUANG Jian1);  GOU Xuejian1)
1)Department of Orthopaedics, Haikou Hospital of Traditional Chinese Medicine, Haikou 570000 2)Department Ⅱ of Orthopaedics, the Second Affiliated Hospital,Hainan Medical College, Haikou 570000
关键词:
miR-424-5p;  PHD2;  骨性关节炎;  凋亡;  炎症因子
Keywords:
miR-424-5p;  PHD2;  osteoarthritis;  apoptosis;  inflammatory factor
分类号:
R684.3
摘要:
目的:研究miR-424-5p对骨性关节炎软骨细胞凋亡、炎症因子分泌的影响及可能调控机制。方法:采用qRT-PCR法检测成人软骨细胞HC、成人骨性关节炎软骨细胞HC-OA中miR-424-5p、PHD2 mRNA的表达,Western blot法检测PHD2蛋白的表达。应用双荧光素酶报告实验检测miR-424-5p与PHD2的靶向关系。将HC-OA分为miR-NC组(转染miR-NC)、miR-424-5p组(转染miR-424-5p模拟物),采用qRT-PCR法检测miR-424-5p的表达,Annexin V-FITC/PI双染法检测细胞凋亡,用ELISA检测细胞培养液上清中IL-6、IL-1β、IL-17的含量,同上检测PHD2 mRNA和蛋白的表达。另取HC-OA,分为miR-424-5p组、miR-424-5p+pcDNA组和miR-424-5p+pcDNA-PHD2组,分别检测细胞中miR-424-5p的表达、细胞凋亡情况及细胞培养液上清中IL-6、IL-1β、IL-17的含量。结果:与HC相比,HC-OA中miR-424-5p表达降低,PHD2 mRNA和蛋白的表达升高(P<0.05)。miR-424-5p与PHD2基因存在靶向关系。过表达miR-424-5p能够抑制HC-OA细胞的凋亡,炎症因子IL-6、IL-1β、IL-17的分泌,及PHD2 mRNA和蛋白的表达(P<0.05)。过表达PHD2可降低HC-OA细胞中miR-424-5p的表达,促进细胞凋亡和炎症因子IL-6、IL-1β、IL-17的分泌(P<0.05)。结论:miR-424-5p可通过靶向PHD2抑制关节炎软骨细胞的凋亡和炎症因子的分泌。
Abstract:
Aim:To investigate the regulatory mechanism of miR-424-5p on osteoarthritis chondrocyte apoptosis and inflammatory factor secretion.Methods:qRT-PCR was used to detect the expressions of miR-424-5p and PHD2 mRNA in adult chondrocytes(HC)and adult osteoarthritis chondrocytes(HC-OA),and Western blot was used to detect the expression of PHD2 protein. Dual luciferase reporter assay was adopted to detect the targeting relationship between miR-424-5p and PHD2.HC-OA was allocated into miR-NC group(transfected miR-NC)and miR-424-5p group(transfected miR-424-5p mimics).qRT-PCR was used to detect the expression of miR-424-5p,Annexin V-FITC/PI double staining was used to detect cell apoptosis,ELISA was used to determine the contents of IL-6, IL-1β and IL-17 in medium supernatant,and the expressions of PHD2 mRNA and protein were also detected.HC-OA was allocated into miR-424-5p group, miR-424-5p+pcDNA group(co-transfected miR-424-5p mimics and pcDNA), and miR-424-5p+pcDNA-PHD2 group(co-transfected miR-424-5p mimics and pcDNA-PHD2).The expression of miR-424-5p, cell apoptosis and the contents of IL-6, IL-1β and IL-17 in medium supernatant were detected.Results:Compared with HC, the expression of miR-424-5p was reduced,while that of PHD2 was increased in HC-OA(P<0.05). There was targeting relationship between miR-424-5p and PHD2.Overexpression of miR-424-5p could significantly inhibit the apoptosis of HC-OA and the secretion of IL-6, IL-1β and IL-17, and negatively regulate the expression of PHD2 in HC-OA cells(P<0.05).Overexpression of PHD2 could significantly reverse the expression of miR-424-5p,induce cell apoptosis, and the secretion of IL-6, IL-1β, and IL-17 by HC-OA cells(P<0.05).Conclusion:miR-424-5p could inhibit the apoptosis and inflammatory factor secretion of osteoarthritic chondrocytes by targeting PHD2.

参考文献/References:

[1] KARDES S, KARAGÜLLE M, GEÇMEN Ì, et al.Outpatient balneological treatment of osteoarthritis in older persons: a retrospective study[J].Z Gerontol Geriatr,2019,52(2):164
[2] 张晨,马骏,吕雷锋, 等.盐酸氨基葡萄糖对膝骨关节炎膝关节软骨下骨Ⅰ型胶原及骨钙素表达的影响[J].郑州大学学报(医学版),2017,52(1):9
ZHANG C, MA J, LYU LF,et al.Effect of glucosamine hydrochloride on typeⅠcollagen and osteocalcin level in knee subchondral bone of patients with thosteoarthritis[J].Journal of Zhengzhou University(Medical Sciences),2017,52(1):9
[3] 熊金河,吴霞,苟玉潇.miR-451对类风湿关节炎患者外周血单个核细胞增殖及炎症反应的影响[J].郑州大学学报(医学版),2019,54(5):689
XIONG JH,WU X,GOU YX.Effects of miR-451 on proliferation and inflammation of peripheral blood mononuclear cells from patients with rheumatoid arthritis[J].Journal of Zhengzhou University(Medical Sciences),2019,54(5):689
[4] 宋小莉,苏娟,刘重阳, 等.类风湿关节炎滑膜组织中脯氨酸羟化酶及希佩尔林道肿瘤抑制蛋白的表达及意义[J].中国组织工程研究,2019,23(23):3623
SONG XL, SU J, LIU CY,et al.Expression and significance of prolyl hydroxylase and von-Hippel-Lindau tumor suppressor protein in the synovial tissues of rheumatoid arthritis[J]. Chinese Journal of Tissue Engineering Research,2019,23(23):3623
[5] APPELHOFF RJ, TIAN YM, RAVAL RR, et al.Differential function of the prolyl hydroxylases PHD1, PHD2, and PHD3 in the regulation of hypoxia-inducible factor[J].J Biol Chem,2004,279(37):38458
[6] KOPAN SKA M, SZALA D, CZECH J, et al.Erratum to: MiRNA expression in the cartilage of patients with osteoarthritis[J].J Orthop Surg Res,2017,12(1):90
[7] 高杰,韩建伟,朱洪繁, 等.miR-424在人骨髓间充质干细胞分化过程中的表达[J].中华创伤杂志,2010,26(8):752
GAO J,HAN JW,ZHU HF, et al.Expressions of miR-424 during differentiation of human bone marrow-derived mesenchymal stem cells[J].Chinese Journal of Trauma,2010,26(8):752
[8] XIA ZN, MA P, WU N, et al.Altered function in cartilage derived mesenchymal stem cell leads to OA-related cartilage erosion[J].Am J Transl Res,2016,8(2):433
[9] WANG P, DONG R, WANG B, et al.Genome-wide microRNA screening reveals miR-582-5p as a mesenchymal stem cell-specific microRNA in subchondral bone of the human knee joint[J].J Cell Physiol,2019,234(12):21877
[10]RAUNER M, FRANKE K, SINGH RP, et al.Bone homeostasis is regulated by the PHD2-HIF2 alpha-EPO axis[J].Amer J Hemat, 2015, 90(8): E159
[11]MUZ B, LARSEN H, MADDEN L, et al.Prolyl hydroxylase domain enzyme 2 is the major player in regulating hypoxic responses in rheumatoid arthritis[J].Arthritis Rheum,2012,64(9):2856
[12]CHENG SH, POURTEYMOOR S, ALARCON C, et al.Conditional deletion of the Phd2 gene in articular chondrocytes accelerates differentiation and reduces articular cartilage thickness[J].Sci Rep, 2017,7:45408
[13]LINDSEY RC, CHENG S, MOHAN S.Vitamin C effects on 5-hydroxymethylcytosine and gene expression in osteoblasts and chondrocytes: potential involvement of PHD2[J].PLoS One,2019,14(8):e0220653

备注/Memo

备注/Memo:
【基金项目】海南省自然科学基金面上项目(817MS156)
【作者简介】陆志夫,通信作者,男,1979年12月生,硕士,副主任医师,研究方向:骨关节病,骨关节损伤,E-mail:lzf888666123@163.com
更新日期/Last Update: 2021-02-15