Dynamic compression counteracts IL-1beta induced inducible nitric oxide synthase and cyclo-oxygenase-2 expression in chondrocyte/agarose constructs.
Dynamic compression counteracts IL-1beta induced inducible nitric oxide synthase and cyclo-oxygenase-2 expression in chondrocyte/agarose constructs.
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DOI：
10.1186/ar2389
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发表时间：
2008
影响因子：
4.9
通讯作者：
Lee DA
中科院分区：
文献类型：
--
作者：
Chowdhury TT;Arghandawi S;Brand J;Akanji OO;Bader DL;Salter DM;Lee DA
一氧化氮和前列腺素E2 （pge2）在骨关节炎的发病机制和关节软骨的分解代谢过程中起关键作用。这些介质受到IL-1β和机械负荷的影响，并涉及诱导型一氧化氮合酶（iNOS）和环加氧酶(COX)-2酶的改变。为了确定两种刺激激活的特定相互作用，我们研究了动态压缩对iNOS和COX-2表达水平的影响，以及p38丝裂原活化蛋白激酶（MAPK）途径的参与。软骨细胞/琼脂糖构建物在有或没有IL-1β和/或SB203580 （p38 MAPK抑制剂）的自由肿胀条件下培养长达48小时。使用完全表征的生物反应器系统，构建体在类似处理下进行动态压缩6,12和48小时。western blotting分析IL-1β和/或SB203580对p38 MAPK的激活或抑制作用。利用实时定量PCR结合分子信标检测iNOS、COX-2、聚集蛋白和II型胶原蛋白信号。用生化法测定亚硝酸盐和PGE2的释放量。采用双向方差分析和事后bonferroni校正t检验对数据进行检验。IL-1β激活p38 MAPK的磷酸化，SB203580消除了这一作用。IL-1β诱导iNOS表达瞬间升高，刺激亚硝酸盐释放。动态压缩或SB203580刺激均可降低IL-1β诱导的iNOS表达和亚硝酸盐的产生。然而，动态压缩和SB203580共同刺激抑制了细胞因子诱导的iNOS表达水平和亚硝酸盐的产生。IL-1β诱导COX-2表达的短暂增加，并刺激PGE2的累积释放。动态压缩或SB203580抑制了这些效应。动态压缩和SB203580共同刺激恢复细胞因子诱导的聚集蛋白表达抑制。这与II型胶原形成对比，在II型胶原中，我们观察到细胞因子和/或SB203580没有反应。这些数据表明，动态压缩直接影响iNOS和COX-2的表达水平。这些分子是目前药理干预的靶点，提高了综合药理和生物物理疗法治疗软骨关节疾病的可能性。
Nitric oxide and prostaglandin E2 (PGE2play pivotal roles in both the pathogenesis of osteoarthritis and catabolic processes in articular cartilage. These mediators are influenced by both IL-1β and mechanical loading, and involve alterations in the inducible nitric oxide synthase (iNOS) and cyclo-oxygenase (COX)-2 enzymes. To identify the specific interactions that are activated by both types of stimuli, we examined the effects of dynamic compression on levels of expression of iNOS and COX-2 and involvement of the p38 mitogen-activated protein kinase (MAPK) pathway. Chondrocyte/agarose constructs were cultured under free-swelling conditions with or without IL-1β and/or SB203580 (inhibitor of p38 MAPK) for up to 48 hours. Using a fully characterized bioreactor system, constructs were subjected to dynamic compression for 6, 12 and 48 hours under similar treatments. The activation or inhibition of p38 MAPK by IL-1β and/or SB203580 was analyzed by western blotting. iNOS, COX-2, aggrecan and collagen type II signals were assessed utilizing real-time quantitative PCR coupled with molecular beacons. Release of nitrite and PGE2 was quantified using biochemical assays. Two-way analysis of variance and the post hoc Bonferroni-corrected t-test were used to examine data. IL-1β activated the phosphorylation of p38 MAPK and this effect was abolished by SB203580. IL-1β induced a transient increase in iNOS expression and stimulated the production of nitrite release. Stimulation by either dynamic compression or SB203580 in isolation reduced the IL-1β induced iNOS expression and nitrite production. However, co-stimulation with both dynamic compression and SB203580 inhibited the expression levels of iNOS and production of nitrite induced by the cytokine. IL-1β induced a transient increase in COX-2 expression and stimulated the cumulative production of PGE2 release. These effects were inhibited by dynamic compression or SB203580. Co-stimulation with both dynamic compression and SB203580 restored cytokine-induced inhibition of aggrecan expression. This is in contrast to collagen type II, in which we observed no response with the cytokine and/or SB203580. These data suggest that dynamic compression directly influences the expression levels of iNOS and COX-2. These molecules are current targets for pharmacological intervention, raising the possibility for integrated pharmacological and biophysical therapies for the treatment of cartilage joint disorders.
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DOI：
10.1097/01.mco.0000068964.34812.2b
发表时间：
2003-05-01
影响因子：
3.1
作者：
Deschner, J;Hofman, CR;Agarwal, S
通讯作者：
Agarwal, S
影响因子：
4.8
作者：
Fitzgerald, JB;Jin, M;Grodzinsky, AJ
通讯作者：
Grodzinsky, AJ
影响因子：
6.9
作者：
De Croos, J. N. A.;Dhaliwal, S. S.;Kandel, R. A.
通讯作者：
Kandel, R. A.
影响因子：
3.5
作者：
Chowdhury, T. T.;Appleby, R. N.;Lee, D. A.
通讯作者：
Lee, D. A.
影响因子：
2.8
作者：
Lee, DA;Bader, DL
通讯作者：
Bader, DL