Mechanical stimulation induces preprotachykinin gene expression in osteoarthritic chondrocytes which is correlated with modulation of the transcription factor neuron restrictive silence factor
Mechanical stimulation induces preprotachykinin gene expression in osteoarthritic chondrocytes which is correlated with modulation of the transcription factor neuron restrictive silence factor
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DOI：
10.1016/j.npep.2008.09.004
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发表时间：
2008-10-01
期刊：
影响因子：
2.9
通讯作者：
Quinn, J. P.
中科院分区：
文献类型：
--
作者：
Howard, M. R.;Millward-Sadler, S. J.;Quinn, J. P.
我们之前已经证明，转录因子神经元限制性沉默因子（NRSF）和截断剪接变体NRSF短形式（sNRSF）是前protachykinin A （TAC1）基因表达的主要调节剂。在这篇文章中，我们讨论了TAC1基因表达是否会在正常和骨关节炎（OA）软骨细胞的机械刺激下受到影响。机械刺激软骨细胞20 min后，在正常组织培养条件下孵育1或3 h，采用RT-PCR和定量PCR （qPCR）检测各时间点TAC1、NRSF和sNRSF mRNA的表达情况。采用Western blotting验证和证实sNRSF蛋白在机械刺激下在软骨细胞中的表达。我们观察到TAC1在正常软骨细胞中表达，没有NRSF或sNRSF表达的证据。机械刺激后正常细胞TAC1 mRNA表达无明显变化。CIA软骨细胞表达TAC1和sNRSF mRNA，但不表达NRSF，机械刺激后TAC1和sNRSF mRNA均显著上调，刺激后3小时恢复到基线水平。sNRSF蛋白在OA软骨细胞刺激后1和2小时上调。总之，OA软骨细胞中TAC1和sNRSF的差异表达与疾病有关。在OA而非正常软骨细胞中，机械刺激后sNRSF和TAC1 mRNA表达的变化提示sNRSF可能参与OA软骨SP生成的调控。正常和OA机械转导反应之间的这些差异可能是病变软骨表型变化产生的重要因素。(C) 2008 Elsevier Ltd版权所有。
We have previously demonstrated that the transcription factor termed neuron restrictive silencer factor (NRSF) and the truncated splice variant, NRSF short form (sNRSF) are major modulators of preprotachykinin A (TAC1) gene expression. In this communication we addressed whether TAC1 gene expression would be effected in response to mechanical stimulation of both normal and osteoarthritic (OA) chondrocytes. Chondrocytes were mechanically stimulated for 20 min, and then incubated under normal tissue culture conditions for 1 or 3 h. RT-PCR and quantitative PCR (qPCR) were used to investigate expression of TAC1, NRSF and sNRSF mRNA at these time points. Western blotting was used to validate and confirm expression of sNRSF protein in chondrocytes in response to mechanical stimulation. We observed that TAC1 was expressed in normal chondrocytes, with no evidence of NRSF or sNRSF expression. TAC1 mRNA expression did not significantly change following mechanical stimulation in normal cells. CIA chondrocytes expressed TAC1 and sNRSF mRNA, though not NRSF, and following mechanical stimulation there was a significant upregulation of both TAC1 and sNRSF mRNA, which returned to baseline levels 3 h post-stimulation. sNRSF protein was upregulated at 1 and 2 h following stimulation of OA chondrocytes.In summary, differential expression of TAC1 and sNRSF in OA chondrocytes associates their expression with the disease. The change in expression of sNRSF and TAC1 mRNA following mechanical stimulation in OA but not normal chondrocytes suggests that sNRSF may be involved in the regulation of SP production in OA cartilage. These differences between normal and OA mechanotransduction responses may be important in the production of phenotypic changes present in diseased cartilage. (C) 2008 Elsevier Ltd. All rights reserved.