Lei Ming, Liu Shi-qing, Liu Yu-Lan
Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan 430060, China.
Acta Pharmacol Sin. 2008 Nov;29(11):1350-6. doi: 10.1111/j.1745-7254.2008.00880.x.
To investigate the effects of resveratrol on interleukin-1beta (IL-1beta) induced catabolism in bone marrow mesenchymal stem cell (MSC) derived chondrocytes cultured on chitosan-gelatin scaffolds (CGS).
The chondrogenesis of alginate-encapsulated MSCs was evaluated by toluidine blue staining, RT-PCR, and immunostaing. MSC-derived chondrocyte morphology cultured on CGS was evaluated by a scanning electron microscope (SEM) and a laser confocal microscope (LCM). When these cells on CGS were pre-stimulated with IL-1beta or co-treated with IL-1beta and resveratrol in the absence and presence of the specific beta1-integrin blocking antibody, collagen type II, aggrecan, matrix metalloproteinase-13 (MMP-13) expression, and the translocation of nuclear factor kappaB (NF-kappaB) were analyzed by Western blot analysis.
Transforming growth factor beta 3 (TGF-beta3) combined with insulin-like growth factor I (IGF-I) induced the cartilage-specific collagen type II, aggrecan expression and extracellular matrix (ECM) accumulation at the end of a 3-week culture. CGS supported those differentiated chondrocytes'attachment, proliferation, migration, and ECM formation. When those cells cultured on CGS were stimulated with IL-beta1 alone, collagen type II and aggrecan expression was inhibited. However, MMP-13 expression increased. Resveratrol reversed the catabolic effects by reducing the translocation of NF-kappaB. A specific beta1-integrin blocking antibody abrogated the effects of resveratrol on IL-1beta stimulated MSC-derived chondrocytes.
These results indicated that resveratrol acts as a NF-kappaB inhibitor to protect MSC-derived chondrocytes on the CGS from the IL-1beta catabolism and these effects are mediated by beta1-integrin.
研究白藜芦醇对白细胞介素-1β（IL-1β）诱导的在壳聚糖-明胶支架（CGS）上培养的骨髓间充质干细胞（MSC）来源软骨细胞分解代谢的影响。
通过甲苯胺蓝染色、逆转录-聚合酶链反应（RT-PCR）和免疫染色评估海藻酸盐包封的MSC的软骨形成。用扫描电子显微镜（SEM）和激光共聚焦显微镜（LCM）评估在CGS上培养的MSC来源软骨细胞的形态。当CGS上的这些细胞在不存在和存在特异性β1整合素阻断抗体的情况下用IL-1β预刺激或与IL-1β和白藜芦醇共同处理时，通过蛋白质印迹分析分析II型胶原、聚集蛋白聚糖、基质金属蛋白酶-13（MMP-13）的表达以及核因子κB（NF-κB）的转位。
在3周培养结束时，转化生长因子β3（TGF-β3）与胰岛素样生长因子I（IGF-I）联合诱导软骨特异性II型胶原、聚集蛋白聚糖表达和细胞外基质（ECM）积累。CGS支持那些分化的软骨细胞的附着、增殖、迁移和ECM形成。当CGS上培养的那些细胞单独用IL-β1刺激时，II型胶原和聚集蛋白聚糖表达受到抑制。然而，MMP-13表达增加。白藜芦醇通过减少NF-κB的转位逆转了分解代谢作用。特异性β1整合素阻断抗体消除了白藜芦醇对IL-1β刺激的MSC来源软骨细胞的作用。
这些结果表明，白藜芦醇作为一种NF-κB抑制剂，保护CGS上的MSC来源软骨细胞免受IL-1β分解代谢的影响，并且这些作用是由β1整合素介导的。
