Cytokines, tumor necrosis factor-α and interleukin-1β, differentially regulate apoptosis in osteoarthritis cultured human chondrocytes
Cytokines, tumor necrosis factor-α and interleukin-1β, differentially regulate apoptosis in osteoarthritis cultured human chondrocytes
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DOI：
10.1016/j.joca.2006.01.005
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发表时间：
2006-07-01
影响因子：
7
通讯作者：
Blanco, F. J.
中科院分区：
文献类型：
--
作者：
Lopez-Armada, M. J.;Carames, B.;Blanco, F. J.
目的：本研究探讨了肿瘤坏死因子-α（TNF-α）和白细胞介素-1 β（IL-1 β）对人软骨细胞凋亡的影响。方法：以放线菌素D（ActD）刺激的骨关节炎（OA）人软骨细胞作为细胞凋亡模型。Caspase家族mRNA表达和蛋白质合成分别通过核糖核酸酶保护试验和Western-blot分析。分别采用MTT法和流式细胞仪检测细胞活力和凋亡。结果：TNF-α和IL-1 β对人软骨细胞caspase mRNA表达的影响存在差异。TNF-α诱导caspase-1和-8 mRNA水平逐渐增加，而IL-1 β则未观察到。TNF-α诱导caspase-3和caspase-7的时间顺序与IL-1 β诱导的时间顺序不同。在培养的软骨细胞中，TNF-α或IL-1 β未改变细胞活力。然后，我们采用ActD作为促进细胞死亡的模型。用TNF-α和ActD处理（TNF-α/ActD）增加了ActD诱导的细胞死亡（23%）。用IL-1 β和ActD处理（IL-1 β/ActD）不调节ActD诱导的细胞死亡。类似地，IL-1 β/ActD不诱导通过与TNF-α/ActD孵育观察到的半胱天冬酶-3和-7活化以及聚（ADP-核糖）聚合酶（PARP）裂解的增加。这些不同的影响不是由于bcl-2或mcl-1水平。吲哚美辛抑制PGE 2合成增加了IL-1 β/Act-D诱导的细胞死亡（59%）。caspase-8抑制剂仅显著降低TNF-α/ActD诱导的细胞死亡（58%）。结论：TNF-α和IL-1 β对人软骨细胞凋亡途径的调节存在差异。这种差异取决于PGE 2和caspase-8水平。(C)2006年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: This study addresses the effects of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) on cell death in human chondrocytesMethods: Osteoarthritis (OA) human chondrocytes stimulated with Actinomycin-D (ActD) were used as a cellular apoptotic model. Caspase family mRNA expression and protein synthesis were analyzed by the ribonuclease protection assay and Western-blot, respectively. Cell viability and apoptosis were evaluated using the 3-[4,5-dimethylthiazol-2yl] 2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry, respectively. Prostaglandin E-2 (PGE(2)) and nitric oxide (NO) were evaluated by enzyme-linked immunosorbent assay (ELISA) and the Griess method, respectively.Results: TNF-alpha and IL-1 beta differentially affected the pattern of caspase mRNA expression by human chondrocytes. TNF-alpha induced a gradual increase in caspase-1 and -8 mRNA levels that was not seen with IL-1 beta. The time sequence of caspase-3 and -7 inductions by TNF-a differs from that induced by IL-1 beta. Cell viability was not modified by TNF-alpha or IL-1 beta in cultured chondrocytes. Then, we employed ActD as a model to facilitate cell death. Treatment with TNF-alpha, and ActD (TNF-alpha/ActD) increased cell death induced by ActD (23%). Treatment with IL-1 beta and ActD (IL-1 beta/ActD) did not modulate ActD-incluced cell death. Similarly, IL-1 beta/ActD did not induce an increase in the activation of caspase-3 and -7 and poly (ADP-ribose) polymerase (PARP) cleavage observed by the incubation with TNF-a/ActD. These different effects were not due to bcl-2 or mcl-1 levels. Inhibition of PGE2 synthesis by indomethacin increased the cell death induced by IL-1 beta/Act-D (59%). An inhibitor of caspase-8 significantly reduced only the TNF-alpha/ActD-incluced cell death (58%).Conclusion: TNF-alpha and IL-1 beta differentially regulate the apoptotic pathway in human chondrocytes. This difference is dependent on PGE2 and caspase-8 levels. (C) 2006 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.