DIO2 modifies inflammatory responses in chondrocytes.
DIO2 modifies inflammatory responses in chondrocytes.
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DOI：
10.1016/j.joca.2012.02.006
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发表时间：
2012-05
影响因子：
7
通讯作者：
Kraus, V. B.
中科院分区：
文献类型：
--
作者：
Cheng, A. W. M.;Bolognesi, M.;Kraus, V. B.
关键词：
硒可中和软骨细胞中白介素 1β (IL-1β) 诱导的炎症反应。我们通过体外敲低原代人软骨细胞中三种主要的硒蛋白：碘甲腺氨酸脱碘酶-2 (DIO2)、谷胱甘肽过氧化物酶-1 (GPX1) 和硫氧还蛋白还原酶-1 (TR1)，研究了其潜在机制。用乱序小干扰 RNA (siRNA) 或对 DIO2、GPX1 和 TR1 具有特异性的 siRNA 转染原代人软骨细胞。 48小时后，将转染的细胞在无血清培养基中培养48小时，最后24小时添加或不添加10 pg/ml IL-1β。 siRNA 的效率通过定量实时聚合酶链式反应 (qRT-PCR) 和蛋白质印迹分析得到证实。通过 qRT-PCR 评估环氧合酶 2 (COX2)、IL-1β 和肝 X 受体 (LXR) α 和 β 的基因表达，以确定硒蛋白敲低对软骨细胞炎症反应的影响。特异性 siRNA 显着降低了 DIO2、GPX1 和 TR1 的 mRNA 表达（分别降低 56%，p=0.0004；96%，p<0.0001；和 66%，p<0.0001）。抑制 DIO2（而非 GPX1 或 TR1）显着增加（约 2 倍）基础（p=0.0005）和 IL-1β 诱导（p <0.0001）COX2 基因表达。同样，抑制 DIO2 显着增加（约 9 倍）IL-1β 诱导的 IL-1β 基因表达（p=0.0056），并导致 LXRα 基因表达减少 32%（p=0.0044），但对 LXRβ 没有影响。硒蛋白 DIO2 的抑制导致强烈的促炎作用，炎症介质 IL-1β 和 COX2 的表达增加，LXRα 的表达减少，表明这可能是介导 DIO2 抗炎作用的上游靶点。
Selenium neutralizes interleukin-1β (IL-1β) induced inflammatory responses in chondrocytes. We investigated potential mechanisms for this through in vitro knockdown of three major selenoproteins, Iodothyronine Deiodinase-2 (DIO2), Glutathione Peroxidase-1 (GPX1), and Thioredoxin Reductase-1 (TR1) in primary human chondrocytes. Primary human chondrocytes were transfected with scrambled small interfering RNA (siRNA) or siRNA specific for DIO2, GPX1 and TR1. After 48 hours, transfected cells were cultured in serum free media for 48 hours, with or without 10 pg/ml IL-1β for the final 24 hours. The efficiency of siRNAs was confirmed by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot analysis. The gene expression, by qRT-PCR, of cyclooxygenase-2 (COX2), IL-1β, and Liver X receptor (LXR) alpha and beta was evaluated to determine the impact of selenoprotein knockdown on inflammatory responses in chondrocytes. The mRNA expression of DIO2, GPX1, and TR1 was significantly decreased by the specific siRNAs (reduced 56%, p=0.0004; 96%, p<0.0001; and 66%, p<0.0001 respectively). Suppression of DIO2, but not GPX1 or TR1, significantly increased (~2-fold) both basal (p=0.0005) and IL-1β induced (p <0.0001) COX2 gene expression. Similarly, suppression of DIO2 significantly increased (~9-fold) IL-1β induced IL-1β gene expression (p=0.0056) and resulted in a 32% (p=0.0044) decrease in LXRα gene expression but no effect on LXRβ. Suppression of the selenoprotein DIO2 resulted in strong pro-inflammatory effects with increased expression of inflammatory mediators, IL-1β and COX2, and decreased expression of LXRα suggesting that this may be the upstream target through which the anti-inflammatory effects of DIO2 are mediated.
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DOI：
10.1083/jcb.93.3.743
发表时间：
1982-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kuettner KE;Pauli BU;Gall G;Memoli VA;Schenk RK
通讯作者：
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影响因子：
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