Epigallocatechin-3-gallate suppresses the global interleukin-1beta-induced inflammatory response in human chondrocytes.
Epigallocatechin-3-gallate suppresses the global interleukin-1beta-induced inflammatory response in human chondrocytes.
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DOI：
10.1186/ar3368
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发表时间：
2011-06-17
影响因子：
4.9
通讯作者：
Haqqi TM
中科院分区：
文献类型：
--
作者：
Akhtar N;Haqqi TM
表没食子儿茶素没食子酸酯（Epigallocatechin-3-gallate，EGCG）是绿色茶中的一种生物活性多酚，通过抑制信号传导和基因表达发挥抗炎作用。白细胞介素-1 β（IL-1β）是与骨关节炎（OA）中软骨降解相关的主要细胞因子。本研究的目的是评估EGCG对IL-1β诱导的人软骨细胞中与OA发病机制相关的蛋白质表达的总体影响。用EGCG（10至100 μ M）预处理原代OA软骨细胞，然后用IL-1β（5 ng/ml）刺激24小时。将培养物上清液与细胞因子抗体阵列一起孵育，并通过增强的荧光观察免疫反应性蛋白质（80种蛋白质）。通过真实的时间PCR验证EGCG对IL-1β诱导的18个选定基因表达的影响，并使用特异性ELISA测定对IL-6、IL-8和肿瘤坏死因子-α（TNF-α）产生的影响。Western免疫印迹法分析EGCG对IL-1β刺激的软骨细胞中白细胞介素-1受体相关激酶1（IRAK-1）和肿瘤坏死因子受体相关因子6（TRAF-6）蛋白的影响。通过使用NF- κB（MG 132）和MAPK（p38-MAPK，SB 202190; JNK-MAPK，SP 600125，ERK-MAPK，PD 98059）的特异性抑制剂，确定核因子κ-B（NF-κB）和促分裂原活化蛋白激酶（MAPK）在所选基因的调节中的作用以及参与EGCG介导的这些基因的调节的机制。在阵列上存在的80种蛋白质中，14%的蛋白质的组成型表达被表没食子酸酯（EGCG）处理改变。对软骨合成代谢反应相关蛋白无明显刺激作用。用IL-1β刺激增强了29种蛋白质的表达。结果表明，EGCG可抑制IL-1β上调的所有29种蛋白的表达。在50和100 μ M浓度下，EGCG也抑制信号中间体TRAF-6的表达（P < 0.05）。我们的研究结果确定了EGCG的几个新靶点，包括上皮中性粒细胞活化肽-78（ENA-78）、粒细胞巨噬细胞集落刺激因子（GM-CSF）、生长相关癌基因（GRO）、GRO-α、IL-6、IL-8、单核细胞趋化蛋白-1（MCP-1）、MCP-3、巨噬细胞炎性蛋白-1 β（MIP-1β）、粒细胞趋化蛋白-2（GCP-2）、MIP-3 α、干扰素-γ-诱导蛋白-10（IP-10）、核小体组装蛋白-2（NAP-2）和白血病抑制因子（LIF）。EGCG的抑制作用主要是通过抑制人软骨细胞NF-κB和c-Jun N末端激酶（JNK）-MAPK的活化来实现的。我们的研究结果表明，表没食子儿茶素没食子酸酯在OA治疗/预防中的潜力可能与其全面抑制人类软骨细胞中的炎症反应的能力有关。这些结果确定了EGCG的其他新靶点，并主张EGCG可能是OA中有效的软骨保护剂。
Epigallocatechin-3-gallate (EGCG) is a bioactive polyphenol of green tea and exerts potent anti-inflammatory effects by inhibiting signaling events and gene expression. Interleukin-1beta (IL-1β) is the principal cytokine linked to cartilage degradation in osteoarthritis (OA). The objective of this study was to evaluate the global effect of EGCG on IL-1β-induced expression of proteins associated with OA pathogenesis in human chondrocytes. Primary OA chondrocytes were pretreated with EGCG (10 to 100 uM) and then stimulated with IL-1β (5 ng/ml) for 24 hours. Culture supernatants were incubated with cytokine antibody arrays and immunoreactive proteins (80 proteins) were visualized by enhanced chemiluminiscence. Effect of EGCG on IL-1β-induced expression of 18 selected genes was verified by Real time-PCR and effect on IL-6, IL-8 and tumor necrosis factor-alpha (TNF-α) production was determined using specific ELISAs. Western immunoblotting was used to analyze the effect of EGCG on the interleukin-1 receptor-associated kinase 1 (IRAK-1) and TNF receptor-associated factor 6 (TRAF-6) proteins in IL-1β-stimulated chondrocytes. The role of nuclear factor kappa-B (NF-κB) and mitogen activated protein kinases (MAPKs) in the regulation of selected genes and the mechanism involved in EGCG mediated modulation of these genes was determined by using specific inhibitors for NF- κB (MG132) and MAPKs (p38-MAPK, SB202190; JNK-MAPK, SP600125, ERK-MAPK, PD98059). Out of 80 proteins present on the array, constitutive expression of 14% proteins was altered by EGCG treatment. No significant stimulatory effect was observed on the proteins associated with cartilage anabolic response. Stimulation with IL-1β enhanced the expression of 29 proteins. Expression of all 29 proteins up-regulated by IL-1β was found to be suppressed by EGCG. EGCG also inhibited the expression of the signaling intermediate TRAF-6 at 50 and 100 uM concentrations (P < 0.05). Our results identified several new targets of EGCG, including epithelial neutrophil activating peptide-78 (ENA-78), granulocyte macrophage colony stimulation factor (GM-CSF), growth- related oncogene (GRO), GRO-α, IL-6, IL-8, monocyte chemotactic protein-1 (MCP-1), MCP-3, macrophage inflammatory protein-1beta (MIP-1β), granulocyte chemotactic protein-2 (GCP-2), MIP-3alpha, interferon-gamma-inducible protein-10 (IP-10), nucleosome assembly protein-2 (NAP-2) and leukemia inhibitory factor (LIF). The inhibitory effects of EGCG were mainly mediated by inhibiting the activation of NF-κB and c-Jun N-terminal Kinase (JNK)-MAPK in human chondrocytes. Our results suggest that the potential of EGCG in OA treatment/prevention may be related to its ability to globally suppress the inflammatory response in human chondrocytes. These results identify additional new targets of EGCG and advocate that EGCG may be a potent chondroprotective agent in OA.
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DOI：
10.1124/jpet.103.059220
发表时间：
2004-02-01
影响因子：
3.5
作者：
Ahmed, S;Wang, NZ;Haqqi, TM
通讯作者：
Haqqi, TM
影响因子：
--
作者：
Akhtar, Nahid;Rasheed, Zafar;Ramamurthy, Sangeetha;Anbazhagan, Arivarasu N.;Voss, Frank R.;Haqqi, Tariq M.
通讯作者：
Haqqi, Tariq M.
DOI：
10.1186/ar318
发表时间：
2001
期刊：
Arthritis research
影响因子：
--
作者：
Cook AD;Braine EL;Campbell IK;Rich MJ;Hamilton JA
通讯作者：
Hamilton JA
DOI：
10.1073/pnas.96.8.4524
发表时间：
1999-04-13
影响因子：
11.1
作者：
Haqqi, TM;Anthony, DD;Mukhtar, H
通讯作者：
Mukhtar, H
影响因子：
2.6
作者：
Cooper, R;Morré, DJ;Morré, DM
通讯作者：
Morré, DM