pomegranate extract inhibits the interleukin-1β-induced activation of mkk-3, p38α-mapk and transcription factor runx-2 in human osteoarthritis chondrocytes石榴提取物抑制mkk-3 interleukin-1β-induced激活,p38α-mapk和转录因子runx-2人类骨关节炎软骨细胞.pdfVIP

Rasheed et al. Arthritis Research Therapy 2010, 12:R195 /content/12/5/R195 RESEARCH ARTICLE Open Access Pomegranate extract inhibits the interleukin-1b- induced activation of MKK-3, p38a-MAPK and transcription factor RUNX-2 in human osteoarthritis chondrocytes * Zafar Rasheed, Nahid Akhtar, Tariq M Haqqi Abstract Introduction: 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-1b (IL-1b) 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-1b-induced activation of MKK3/6, p38-MAPK isoforms and the activation of transcription factor RUNX-2 in primary human OA chondrocytes. Methods: Human chondrocytes were derived from OA cartilage by enzymatic digestion, treated with PE and then stimulated with IL-1b. 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. Results: Human OA chondrocytes expressed p38-MAPK isoforms p38a, -g and -δ, but not p38b. IL-1b enhances the phosphorylation of the p38a-MAPK and p38g-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-1b