miR-335-5p靶向程序化细胞死亡基因5对骨关节炎软骨细胞增殖、凋亡的影响  

Effects of mi R-335-5p on proliferation and apoptosis of osteoarthritic chondrocytes by targeting programmed cell death 5

作　　者：张云青 王健 阳春华 李聪 Zhang Yunqing;Wang Jian;Yang Chunhua;Li Cong(Department of Orthopedics,the First Hospital of Changsha,Changsha 410000,Hunan Province,China)

机构地区：[1]长沙市第一医院骨科,湖南省长沙市410000

出　　处：《中国组织工程研究》2021年第14期2142-2147,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:研究发现,miR-335-5p在骨关节炎患者关节液中表达下调,其可能与骨关节炎的发生发展有关。目的:探讨miR-335-5p对骨关节炎软骨细胞增殖和凋亡的影响及潜在机制。方法:分离培养人原代关节软骨细胞,以10μg/L白细胞介素1β处理软骨细胞建立骨关节炎模型,并对处理后的软骨细胞进行转染和分组:对照组(常规培养基)、白细胞介素1β组、mi R-NC组、mi R-335-5p组、si-NC组、si-PDCD5组、mi R-335-5p+pc DNA3.1组、mi R-335-5p+pcDNA3.1-PDCD5组。培养至24,48和72 h采用MTT法和流式细胞术分别检测软骨细胞增殖和凋亡率;qRT-PCR检测软骨细胞miR-335-5p、程序化细胞死亡基因5(programmed cell death 5,PDCD5)、软骨蛋白聚糖抗体(ADAMTS-5)和基质金属蛋白酶13 mRNA的表达;Western blot实验检测软骨细胞中PDCD5、Cyclin D1、p21、Bax和Bcl-2蛋白的表达,双荧光素酶报告系统验证miR-335-5p与PDCD5的调控关系。研究方案的实施符合长沙市第一医院的相关伦理要求。结果与结论:①与对照相比,白细胞介素1β可抑制软骨细胞增殖并促进细胞凋亡,抑制软骨细胞中miR-335-5p的表达;过表达miR-335-5p可促进白细胞介素1β作用的软骨细胞增殖和抑制细胞凋亡;②miR-335-5p靶向负调控PDCD5的表达;③抑制PDCD5可促进白细胞介素1β作用的软骨细胞增殖并抑制细胞凋亡;过表达PDCD5可逆转上调miR-335-5p对白细胞介素1β作用的软骨细胞增殖和凋亡的作用;④结果说明,miR-335-5p通过靶向PDCD5促进细胞增殖和抑制细胞凋亡;miR-335-5p是骨关节炎潜在分子靶点。BACKGROUND:Studies have found that miR-335-5 p is down-regulated in the synovial fluid of patients with osteoarthritis,which may be related to the occurrence and development of osteoarthritis.OBJECTIVE:To investigate the effects of miR-335-5 p on the proliferation and apoptosis of chondrocytes in osteoarthritis and the underlying mechanism.METHODS:Human primary articular chondrocytes were isolated and cultured,and the cells were then treated with 10μg/L interleukin-1βto establish an osteoarthritis model.The treated cells were divided into normal group(normal culture medium),interleukin-1βgroup,miR-NC group,miR-335-5 p group,siNC group,si-programmed cell death 5(PDCD5)group,miR-335-5 p+pcDNA3.1 group,and miR-335-5 p+pcDNA3.1-PDCD5 group.After 24,48,and 72 hours of culture,the cell proliferation and apoptosis rate were detected by MTT and flow cytometry,respectively.The mRNA expression levels of miR-335-5 p,PDCD5,ADAMTS-5 and matrix metalloproteinase-13 in chondrocytes were detected by qRT-PCR.The expression levels of PDCD5,Cyclin D1,p21,Bax and Bcl-2 proteins in chondrocytes were determined by western blot.The relationship between miR-335-5 p and PDCD5 was verified by double luciferase reporting assay system.The study procedures were implemented in line with the relevant ethic requirements of the First Hospital of Changsha.RESULTS AND CONCLUSION:Compared with the control group,interleukin-1βinhibited the cell proliferation and promoted the apoptosis of chondrocytes,and inhibited the expression of miR-335-5 p in chondrocytes.Over-expression of miR-335-5 p promoted the proliferation and inhibited the apoptosis of chondrocytes induced by interleukin-1β.miR-335-5 p targeted and negatively regulated the expression of PDCD5.Inhibition of PDCD5 promoted the proliferation of chondrocytes induced by interleukin-1β.Overexpression of PDCD5 reversed the effects of miR-335-5 p up-regulation on the proliferation and apoptosis of chondrocytes induced by interleukin-1β.To conclude,miR-335-5 p promotes cell proliferation an

关 键 词：骨关节炎 软骨细胞 miR-335-5p PDCD5 增殖 凋亡 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]