NF-κBp65-specific siRNA inhibits expression of genes of COX-2, NOS-2 and MMP-9 in rat IL-1β-induced and TNF-α-induced chondrocytes
NF-κBp65-specific siRNA inhibits expression of genes of COX-2, NOS-2 and MMP-9 in rat IL-1β-induced and TNF-α-induced chondrocytes
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NF-κBp65 特异性 siRNA 抑制大鼠 IL-1β 诱导和 TNF-α 诱导软骨细胞中 COX-2、NOS-2 和 MMP-9 基因的表达
DOI：
10.1016/j.joca.2005.10.009
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发表时间：
2006-04-01
影响因子：
7
通讯作者：
Changlong, Y.
中科院分区：
文献类型：
--
作者：
Lianxu, C.;Hongti, J.;Changlong, Y.
目的：小干扰RNA(SiRNA)在哺乳动物体细胞中触发RNA干扰。核因子-kappaB是一种参与炎症和免疫激活的转录因子。本研究旨在利用核因子-kappa Bp65特异性siRNA抑制与骨关节炎软骨病变发生发展过程平行的环氧合酶-2(COX-2)、一氧化氮合酶-2(NOS-2)和基质金属蛋白酶-9(MMP9)基因在诱导的软骨细胞中的表达，从而探索一种新的骨关节炎的基因治疗方法。方法：采用Western印迹和逆转录聚合酶链式反应(RT-PCR)方法优化核因子-kappa Bp65特异性siRNA对培养的大鼠软骨细胞的沉默作用，并检测其表达的变化。诱导软骨细胞中NOS-2和MMP9的表达。用凝胶迁移率改变分析法(EMSA)检测核因子-kappaB的活性。结果：核因子-kappa Bp65特异性siRNA可抑制IL-1β和肿瘤坏死因子-α诱导的软骨细胞COX-2、NOS-2和MMP-9的表达，抑制核因子-kappaBp65的表达和核因子-kappaB的激活。这表明，核因子-kappa Bp65特异性siRNA有可能成为早期预防和治疗骨性关节炎的有用药物。(C)2005年国际骨性关节炎研究会。爱思唯尔有限公司出版。保留所有权利。
Objective: Small interfering RNA (siRNA) triggers RNA interference in mammalian somatic cells. Nuclear factor kappa B (NF-kappa B) is a transcription factor that is implicated in inflammation and immune activation. This study was to use NF-kappa Bp65-specific siRNA to inhibit the expression of genes of cyclooxygenase-2 (COX-2), nitric oxide synthase-2 (NOS-2) and matrix metalloproteinase-9 (MMP-9), which is paralleled with the initiation and progression of cartilage lesions in osteoarthritis (OA) model, in induced chondrocytes, and therefore to explore a new gene therapy for OA.Methods: Western blot and reverse transcriptase polymerase chain reaction (RT-PCR) were performed to optimize the silencing effects of NF-kappa Bp65-specific siRNA in cultured rat chondrocytes, and then to determine the expression of COX-2, NOS-2 and MMP-9 in induced chondrocytes. The activation of NF-kappa B was determined by electrophoretic mobility shift assay (EMSA). Western blot and RT-PCR were subjected to densitometric analysis and then band intensities were also determined.Results: The NF-kappa Bp65-specific siRNA inhibited the expression of NF-kappa Bp65 and activation of NF-kappa B, reducing significantly the expression of COX-2, NOS-2 and MMP-9 induced by interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) in cultured chondrocytes.Conclusions: NF-kappa Bp65-specific siRNA can inhibit the expression of COX-2, NOS-2 and MMP-9 in IL-1 beta-induced and TNF-alpha-induced chondrocytes. This suggests that NF-kappa Bp65-specific siRNA has potential to be a useful, preventive and therapeutic agent for OA at early stages. (c) 2005 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.