筛选TXNDC5与胰岛素相关信号通路关键基因的探讨  

Screening key genes for TXNDC5 and insulin signaling pathway

作　　者：李健[1,2] 徐冰[2] 闫新峰[3] 徐万菊[4] 常晓天[2] 

机构地区：[1]山东大学医学院,山东济南250012 [2]山东大学附属千佛山医院医学研究中心,山东济南250014 [3]山东大学附属千佛山医院骨科,山东济南250014 [4]山东大学附属千佛山医院检验科,山东济南250014

出　　处：《山东大学学报（医学版）》2017年第3期88-93,共6页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(ZR2014HP018);山东省医药卫生科技发展计划(2013WS0131)

摘　　要：目的旨在探讨硫氧还蛋白5(TXNDC5)是否通过胰岛素抵抗或胰岛素信号通路参与类风湿关节炎(RA)发病过程。方法分离培养RA滑膜细胞(RASFs),用小干扰RNA抑制其TXNDC5的表达,提取各样本总RNA组成样本池,采用胰岛素抵抗PCR Array和胰岛素信号通路PCR Array检测相关基因的表达,采用RT-PCR法和ELISA法分别检测相关基因mRNA和蛋白水平的表达。结果 RASFs中TXNDC5表达被抑制后,PCR Array检测IGF-1、PCK1、SLC2A4、IL1R1和胰岛素样生长因子结合蛋白1(IGFBP1)的表达明显发生变化。经验证,IGFBP1在mRNA和蛋白水平均显著增高,差异有统计学意义(P均<0.001);而IGF-1、PCK1、SLC2A4和IL1R1的表达差异无统计学意义(P=0.792 7,P=0.713 0,P=0.452 9,P=0.354 2)。胰岛素样生长因子结合蛋白3(IGFBP3)在RASFs中的表达无明显变化(P=0.511 5)。结论在RASFs中,TXNDC5可能通过抑制胰岛素信号通路IGFBP1表达参与RA病变过程。Objective To explore the pathogenic mechanism of thioredoxin domain containing protein 5（TXNDC5）involvement in rheumatoid arthritis（RA）.Methods Human rheumatoid arthritis synovial cells（RASFs） were separated from RA patients.After the TXNDC5 expression was inhibited by anti-TXNDC5 siRNA,the total RNA was pooled,and the expressions of related genes were detected with PCR Array.The mRNA and protein expressions of related genes were verified using RT-PCR and ELISA.Results PCR Array analyses demonstrated that IGF-1,PCK1,SLC2A4,IL1 R and IGFBP1 expressions significantly changed after TXNDC5 expression was inhibited.RT-PCR and ELISA verification indicated that IGFBP1 expression was significantly enhanced in both mRNA and protein levels（both P〈0.001）,while expressions of IGF-1,PCK1,SLC2A4 and IL1R1 showed no significant difference（P=0.792 7,P=0.713 0,P=0.452 9,P=0.354 2）.No difference was observed in IGFBP3 expressions（P=0.511 5）.Conclusion In RASFs,TXNDC5 is involved in RA by regulating the expression of IGFBP1.

关 键 词：硫氧还蛋白5 胰岛素样生长因子结合蛋白1 类风湿关节炎 胰岛素抵抗 胰岛素信号通路 

分 类 号：R34[医药卫生—基础医学]