miR-140在不同阶段骨关节炎软骨细胞中的表达规律及功能  

Expression and Function of miR-140 in Different Stage Chondrocytes of Patients with Osteoarthritis

作　　者：李灿锋[1] 陈卓[2] 杨静[1] 周宗科[1] 康鹏德[1] 裴福兴[1] 沈彬[1] 

机构地区：[1]四川大学华西医院骨科,四川成都610041 [2]四川大学华西医院转化神经科学中心,四川成都610041

出　　处：《实用骨科杂志》2017年第11期995-999,共5页Journal of Practical Orthopaedics

基　　金：国家自然科学基金项目(81672219;81601936);四川省科技厅科技支撑计划(2014SZ0023-2)

摘　　要：目的观察人膝关节不同退变阶段骨关节炎(osteoarthritis,OA)软骨细胞中微小RNA-140(micro RNA-140,miR-140)表达规律及转染双链miR-140模拟物/抑制物(double strands-miR-140 mimic/inhibitor,ds-miR-140mimic/inhibitor)对OA软骨细胞功能的影响。方法根据Kellgren and Lawrence X线分级,膝关节软骨分为正常组(O级)、中期OA组(Ⅲ级)和晚期OA组(Ⅳ级)。各组收集软骨标本6~8例,体外分离及培养软骨细胞,采用第2代软骨细胞进行检测。运用实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测各组miR-140相对表达量及Ⅱ型胶原(collagentypeⅡa,COL2A1)及Ⅹ型胶原(Collagentype Xa,COL10A1)的表达情况。向中期OA组软骨细胞转染ds-miR-140mimic/inhibitor后,观察软骨形成因子SRY基因(SRY-related high mobility group box gene9,SOX9),COL2A1及COL10A1表达情况。结果 miR-140在OA软骨细胞中表达显著下降,中、晚期OA组miR-140和正常组相比分别减少78.9%和94.5%(P<0.05),晚期OA组miR-140较中期OA组减少73.9%(P<0.05)。中、晚期OA组SOX9的mRNA较正常组分别减少34.1%和49.3%(P<0.05),COL2A1的mRNA较正常组分别减少36.7%和78.7%(P<0.05),中、晚期OA组COL10A1的mRNA较正常组分别增加1.66倍和2.10倍(P<0.05)。向中期OA组转染ds-miR-140mimic/inhibitor后,SOX9的mRNA分别增加1.95倍和减少59.1%,COL2A1的mRNA分别增加1.87倍和减少68.8%(P<0.05),COL10A1的mRNA分别减少73.0%和增加1.96倍(P<0.05)。结论miR-140在OA软骨细胞中表达显著下降,且随病情加重而进一步减少;转染ds-miR-140 mimic可增加SOX9和COL2A1的表达,同时降低COL10A1的表达。Objective To investigate the expression miR-140 in different stage of osteoarthritis and its function after transfecting ds-miR-140 mimic/inhibitor into chondrocytes.Methods Cartilage was classified into three groups,normal group,mid-staged group and advanced-staged group,according to the Kellgren and Lawrence criterion.6-8 cases of normal or OA cartilage were collected and chondrocytes were isolated.The second generation of chondrocytes were used to test.The levels of miR-140,COL2A1 and COL10A1 mRNA were measured with real-time fluorescent quantitative PCR.The ds-miR-140 mimic/inhibitorwere transfected into the mid-staged group,and the expression of COL2A1,COL10A1 and SOX9 were measured.Results Compared with the normal group,miR-140 expression in the mid-and advanced-staged group was decreased by 34.1% and 49.3% respectively,and its expression in advanced-staged group was decreased by 73.9%.Beside,SOX9 expression in the mid-and advanced-staged group was decreased by 75% and82.7%respectively,COL2A1 expression decreased by 36.7% and 78.7% respectively,while COL10A1 expression increased to 1.66 times and 2.10 times respectively.After transfecting ds-miR-140 mimic/inhibitor into the mid-staged group,SOX9 expression was increased by 1.95 times or decreased by 59.1%,COL2A1 expression was increased by 1.87 times or decreased by 68.8%,and COL10A1 expression was decreased by 73.0% or increased by 1.96 times.Conclusion The expression level of miR-140 decreased in osteoarthritic chondrocytes,and was negatively related to the severity of OA.Overexpression its level by transfecting ds-miR-140 mimic into chondrocytes could up-regulate SOX9 and COL2A1 expression while down-regulate COL10A1 expression.

关 键 词：骨关节炎 软骨细胞 微小RNA140 Ⅱ型胶原 X型胶原 

分 类 号：R684.3[医药卫生—骨科学]