Pomegranate extract inhibits the interleukin-1β-induced activation of MKK-3, p38α-MAPK and transcription factor RUNX-2 in human osteoarthritis chondrocytes.
Pomegranate extract inhibits the interleukin-1β-induced activation of MKK-3, p38α-MAPK and transcription factor RUNX-2 in human osteoarthritis chondrocytes.
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DOI：
10.1186/ar3166
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发表时间：
2010
影响因子：
4.9
通讯作者：
Haqqi TM
中科院分区：
文献类型：
--
作者：
Rasheed Z;Akhtar N;Haqqi TM
历史上，石榴因其药用价值而备受推崇。P38-MAPK是骨关节炎(OA)的主要信号转导途径，IL-1β(IL-1β)激活p38-MAPK在骨关节炎软骨分解代谢的多种介质的表达和产生中起关键作用。在这项研究中，我们检测了标准化石榴提取物(PE)对IL-1β诱导的原代人骨关节炎软骨细胞MKK3/6、p38-MAPK亚型激活和转录因子RUNX-2激活的影响。采用酶消化法从人骨关节炎软骨中分离出软骨细胞，用PE处理后，再用IL-1β刺激。RT-PCR检测p38-MAPK亚型的基因表达。免疫印迹法检测MAPKs的活化情况。免疫沉淀法检测p38-MAPK亚型的激活情况。用高灵敏度、高特异性的酶联免疫吸附试验检测RUNX-2的DNA结合活性。用siRNAs进行药理学研究，以阐明所涉及的途径。人骨关节炎软骨细胞表达p38-MAPK亚型p38-α，-γ和-δ，但不表达p38-β。IL-1β可促进人骨关节炎软骨细胞p38α-MAPK和p38γ-MAPK亚型的磷酸化，但不能促进p38δ-MAPK亚型的磷酸化。人骨关节炎软骨细胞中p38-MAPK的激活主要是通过MKK3的激活来实现的。此外，我们还发现富含多酚的PE抑制IL-1β诱导的MKK3、p38MAPK亚型的激活和转录因子RUNX-2的α结合活性。我们的结果为石榴提取物保护软骨和抑制关节炎作用的分子基础提供了重要的见解。PE对IL-1β刺激的人骨关节炎软骨细胞的这些新的药理作用表明，PE或PE衍生化合物可能被开发为MKK和p38-MAPK抑制剂，用于治疗骨关节炎和其他退行性/炎症性疾病。
Pomegranate has been revered throughout history for its medicinal properties. p38-MAPK is a major signal-transducing pathway in osteoarthritis (OA) and its activation by interleukin-1β (IL-1β) plays a critical role in the expression and production of several mediators of cartilage catabolism in OA. In this study we determined the effect of a standardized pomegranate extract (PE) on the IL-1β-induced activation of MKK3/6, p38-MAPK isoforms and the activation of transcription factor RUNX-2 in primary human OA chondrocytes. Human chondrocytes were derived from OA cartilage by enzymatic digestion, treated with PE and then stimulated with IL-1β. Gene expression of p38-MAPK isoforms was measured by RT-PCR. Western immunoblotting was used to analyze the activation of MAPKs. Immunoprecipitation was used to determine the activation of p38-MAPK isoforms. DNA binding activity of RUNX-2 was determined using a highly sensitive and specific ELISA. Pharmacological studies to elucidate the involved pathways were executed using transfection with siRNAs. Human OA chondrocytes expressed p38-MAPK isoforms p38α, -γ and -δ, but not p38β. IL-1β enhances the phosphorylation of the p38α-MAPK and p38γ-MAPK isoforms but not of p38δ-MAPK isoform in human OA chondrocytes. Activation of p38-MAPK in human OA chondrocytes was preferentially mediated via activation of MKK3. In addition, we also demonstrate that polyphenol rich PE inhibited the IL-1β-induced activation of MKK3, p38α-MAPK isoform and DNA binding activity of the transcription factor RUNX-2. Our results provide an important insight into the molecular basis of the reported cartilage protective and arthritis inhibitory effects of pomegranate extract. These novel pharmacological actions of PE on IL-1β stimulated human OA chondrocytes impart a new suggestion that PE or PE-derived compounds may be developed as MKK and p38-MAPK inhibitors for the treatment of OA and other degenerative/inflammatory diseases.
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影响因子：
4.9
作者：
Rasheed Z;Anbazhagan AN;Akhtar N;Ramamurthy S;Voss FR;Haqqi TM
通讯作者：
Haqqi TM
DOI：
10.1124/jpet.109.165209
发表时间：
2010-05-01
影响因子：
3.5
作者：
Rasheed, Zafar;Akhtar, Nahid;Haqqi, Tariq M.
通讯作者：
Haqqi, Tariq M.
影响因子：
4.8
作者：
Enslen, H;Raingeaud, J;Davis, RJ
通讯作者：
Davis, RJ
影响因子：
5.1
作者：
Loeser RF;Erickson EA;Long DL
通讯作者：
Long DL
影响因子：
14.9
作者：
Mengshol, JA;Vincenti, MP;Brinckerhoff, CE
通讯作者：
Brinckerhoff, CE