【摘要】 目的探讨基因Wip1异常表达与椎间盘退行性疾病(intervertebral disc degeneration,IDD)的关系。方法选取62例患者退行性病变的椎间盘组织,通过实时定量PCR测定Wip1基因表达量;siRNA干扰人髓核(nucleus pulposus,NP)细胞Wip1的表达后,通过SA-β-ga1染色检测细胞衰老情况;通过MTT检测NP细胞的增殖能力;通过实时定量PCR检测NP细胞衰老分子标记的表达水平;γ射线照射siRNA转染后的NP P3细胞,分别于不同时间点行SA-β-gal染色检查。结果 62例病变椎间盘组织中,18例Wip1基因的表达明显下降(P<0.01);siRNA干扰人NP细胞Wip1表达后,衰老细胞数量明显增加;MTT结果表明,干扰Wip1的表达能够显著抑制人NP细胞的增殖;P3细胞培养8 d,实时定量PCR检测发现,Col2a1、Agc、Ver的表达水平显著下调,MMP3的表达水平明显增高。y射线照射NP细胞后,SA-β-gal染色显示,Wip1表达组出现明显的衰老现象。结论 Wip1的低表达与椎间盘退行性病变的发生相关,干扰Wip1的表达能够促进NP细胞衰老,而过表达Wip1可以有效抑制细胞衰老的发生。更多还原

【Abstract】 Objective To investigate the association between aberrant expression of Wip1 and intervertebral disc degeneration(IDD).Methods The expression of Wip1 was examined by qRTPCR in 62 discopathy tissue samples.Cell senescence and proliferation was evaluated by SA-β-gal and MTT when Wip1 was knocked down by siRNA interference of in nucleus pulposus(NP) cells. Senescence marker expression was detected by qRT-PCR.The NP cells treated by Wip1 siRNA were exposed toγradiation and then evaluated for cell senescence by SA-p-gal at different time points. Results The expression of Wip1 gene was down-regulated in 18/62 discopathy tissue samples compared with normal tissues.Moreover,the depletion of Wip1 by RNA interference(RNAi) significandy increased NP cell senescence.MTT analysis showed that knockdown of Wip1 markedly inhibited the proliferation of NP cells.In addition,qRT-PCR data indicated that the expression of Col2al,Age and Ver were down-regulated,while MMP3 was up-regulated after 8 days of P3 cell cultured.SA-β-gal staining showed that expression of Wip1 induced NP cell senescence significantly when cells exposed to y radiation.Conclusion The aberrant expression of Wip1 is associated with discopathy.Knockdown of Wip1 may increase senescence of NP cells,while overexpression of Wip1 may inhibit pathogenesis of discopathy.更多还原