miR-17在类风湿关节炎发病中靶向信号转导与转录活化因子3调控滑膜成纤维细胞增殖和炎症因子释放的研究  

The reaserch about miR-17 targeting signal transducer and activator of transcription 3 to regulate the proliferation of rheumatoid arthritis synovial fibroblasts and the release of inflammatory factors from rheumatoid arthritis synovial fibroblasts in the pathogenesis of rheumatoid arthritis

作　　者：王友庆[1] 陈士芳[1] 梅珏[1] Wang Youqing;Cheng Shifang;Mei Jue(Department of Rheumatology,Huzhou Central Hospital,Zhejiang 313000,China)

机构地区：[1]浙江省湖州市中心医院风湿免疫科,313000

出　　处：《中华风湿病学杂志》2019年第5期314-319,I0002,共7页Chinese Journal of Rheumatology

摘　　要：目的探讨微RNA(miR)-17是否在调控信号转导与转录活化因子3(STAT3)表达、影响滑膜成纤维细胞(RASFs)增殖和炎症因子释放中起作用。方法采集RA患者滑膜组织,另收集OA患者滑膜组织作对照,检测miR-17、STAT3、p-STAT3表达并用Mann-Whitney U检验进行统计分析。分析IL-17A处理对RASFs细胞miR-17、STAT3的mRNA表达、细胞增殖及IL-6、IL-8分泌量的影响。通过细胞转染miR-17mimic及siRNA-STAT3,分析其对细胞增殖及IL-6、IL-8分泌量的影响。采用两独立样本t检验对RASFs细胞实验结果进行统计学分析。结果与OA患者相比,RA患者滑膜组织miR-17表达明显降低(Mann-Whitney U=6,P<0.01),STAT3表达明显升高(Mann-Whitney U=32,P<0.01)。OA患者关节腔滑液IL-17A、IL-6和IL-8的表达分别为[(53±12)pg/ml、(43±9)pg/ml和(33±5)pg/ml],低于RA患者[(170±30)pg/ml、(222±37)pg/ml和(156±34)pg/ml,t=18.83,24.28,19.23,P均<0.01]。IL-17A处理明显下调miR-17表达[(1.00±0.12)与(0.37±0.04),t=8.63,P<0.01]、上调STAT3表达[(1.00±0.14)与(1.92±0.23),t=5.92,P<0.01]、促进RASFs细胞增殖、促进炎症因子IL-6、IL-8释放。双荧光素酶报告基因实验结果证实,miR-17与STAT3之间存在靶向调控关系。转染miR-17mimic或siRNA-STAT3可明显下调RASFs细胞STAT3、p-STAT3表达,抑制细胞增殖(miR-NC与miR-17mimic为[(26.9±2.8)与(41.5±3.1),t=6.06,P<0.01];siRNA-NC与siRNA-STAT3为[(23.5±2.4)与(43.2±3.2),t=8.58,P<0.01],使炎症因子[IL-6(miR-NC与miR-17mimic为(110±13)与(66±9),t=4.88,P<0.01];siRNA-NC与siRNA-STAT3为[(117±12)与(70±6),t=6.10,P<0.01]、IL-8[miR-NC与miR-17mimic为(127±10)与(72±7),t=8.10,P<0.01];siRNA-NC与siRNA-STAT3为[(123±11)与(52±6),t=10.19,P<0.01]分泌明显减少。结论miR-17表达降低可能在升高STAT3表达和促进RA发病中起作用,过表达miR-17则具有抑制STAT3表达,减弱RASFs细胞增殖和炎症因子释放的作用。Objective To investigate whether miR-17 plays a role in re-gulation of signal transducer and activator of transcription 3 (STAT3) expression, affecting the proliferation of rheumatoid arthritis synovial fibroblasts (RASFs) and the release of inflammatory factors from RASFs. Methods The synovial tissues of rheumatoid arthritis (RA) patients were collected. As a comparison, synovial tissues from osteoarthritis(OA) patients were collected as control, the expression of miR-17, STAT3 were detected. The results were analyzed by Mann-Whitney U test. To analyze the effects of interleukin (IL)-17A treatment on the expression of micro RNA-17 and STAT3, cell proliferation and secretion of IL-6 and IL-8 in RASFs cells. The effects of cell proliferation and secretion of IL-6 and IL-8 were analyzed by cell transfection of micro RNA-17 mimic and siRNA-STAT3. The results about RASFs cells were analyzed by t-testof two independent samples. Results Compared with OA patients, the expression of miR-17 in synovial tissues of RA patients decreased significantly (Mann-Whitney U=6, P<0.01), while the expression of STAT3 increased obviously (Mann-Whitney U=32, P<0.01). The expressions of IL-17A, IL-6 and IL-8 in synovial fluid of patients with OA were (53±12),(43±9) and (33±5), respectively, significantly lower than those of patients with RA [(170±30),(222±37) and (156±34), t=18.83, 24.28, 19.23, P<0.01]. IL-17A treatment significantly lowered the expression of miR-17[(1.00±0.12) vs (0.37±0.04), t=8.63, P<0.01], while up-regulated the expression of STAT3 [(1.00±0.14) vs(1.92±0.23), t=5.92, P<0.01), promoted the proliferation of RASFs cells, and promoted the release of inflammatory factors IL-6 and IL-8. The results of the double luciferase reporter gene showed that there was a targeting regulation relationship between miR-17 and STAT3. Transfection of miR-17 mimic or siRNA-STAT3 could significantly reduce the expression of STAT3 and p-STAT3 in RASFs cells, along with the inhabitation of cell proliferation [miR-NC vs miR-17 mimi

关 键 词：关节炎 类风湿 miR-17 STAT3 滑膜成纤维细胞 细胞增殖 

分 类 号：R593.22[医药卫生—内科学]