Dynamic compression influences interleukin-1β-induced nitric oxide and prostaglandin E2 release by articular chondrocytes via alterations in iNOS and COX-2 expression
Dynamic compression influences interleukin-1β-induced nitric oxide and prostaglandin E2 release by articular chondrocytes via alterations in iNOS and COX-2 expression
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DOI：
10.3233/bir-2008-0474
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发表时间：
2008-01-01
期刊：
影响因子：
1.1
通讯作者：
Lee, D. A.
中科院分区：
文献类型：
--
作者：
Chowdhury, T. T.;Akanji, O. O.;Lee, D. A.
白细胞介素-1 β（IL-1 β）诱导软骨细胞释放一氧化氮（（NO）-N-中心点）和前列腺素E-2（PGE（2）），这种作用可以通过应用动态压缩逆转。以往的研究表明，整合素可能发挥作用。另外，IL-1 β通过上游p38 MAPK激活上调iNOS和考克斯-2 mRNA的表达。将牛软骨细胞接种在琼脂糖构建体中，并与0或10个（-1）IL-1 β一起培养，在1 Hz下施加或不施加15%动态压缩应变。ng.ml选择的抑制剂被用于询问α 5 β 1整合素信号传导和p38 MAPK活化在介导响应于IL-1 β和动态压缩的（NO）-N-中心点和PGE（2）的释放中的作用。采用实时定量PCR检测iNOS和考克斯-2的相对表达水平。使用Griess分析测量（NO）-N-中心点的稳定终产物亚硝酸盐，使用酶免疫分析测量PGE 2释放。IL-1 β促进（NO）-N-中心点和PGE 2释放，并且这种作用被动态压缩的应用逆转。与整合素结合肽（GRGDSP）共孵育消除了压缩诱导的效果。实时荧光定量PCR分析显示，IL-1 β可增强iNOS和考克斯-2 mRNA的表达，在6或12 h表达最高。动态压缩通过p38 MAPK敏感通路降低这种效应。这些结果表明，动态压缩通过直接影响iNOS和考克斯-2的表达水平来消除（NO）-N-中心点和PGE（2）的释放。
Interleukin-1 beta(IL-1 beta) induces the release of nitric oxide ((NO)-N-center dot) and prostaglandin E-2 (PGE(2)) by chondrocytes and this effect can be reversed with the application of dynamic compression. Previous studies have indicated that integrins may play a role. In addition, IL-1 beta upregulates the expression of iNOS and COX-2 mRNA via upstream activation of p38 MAPK. The current study examines the involvement of these pathways in mediating (NO)-N-center dot and PGE2 release in IL-1 beta stimulated bovine chondrocytes subjected to dynamic compression.Bovine chondrocytes were seeded in agarose constructs and cultured with 0 or 10 ng.ml(-1) IL-1 beta with or without the application of 15% dynamic compressive strain at 1 Hz. Selected inhibitors were used to interrogate the role of alpha 5 beta 1 integrin signalling and p38 MAPK activation in mediating the release of (NO)-N-center dot and PGE(2) in response to both IL-1 beta and dynamic compression. The relative expression levels of iNOS and COX-2 were assessed using real-time quantitative PCR. Nitrite, a stable end product of (NO)-N-center dot, was measured using the Griess assay and PGE2 release was measured using an enzyme immunoassay.IL-1 beta enhanced (NO)-N-center dot and PGE(2) release and this effect was reversed by the application of dynamic compression. Co-incubation with an integrin binding peptide (GRGDSP) abolished the compression-induced effect. Real-time quantitative PCR analysis revealed that IL-1 beta enhanced iNOS and COX-2 mRNA levels, with the maximum expression at 6 or 12 hours. Dynamic compression reduced this effect via a p38 MAPK sensitive pathway. These results suggest that dynamic compression acts to abrogate of (NO)-N-center dot and PGE(2) release by directly influencing the expression levels of iNOS and COX-2.