中文摘要：

目的研究白藜芦醇对骨髓间充质干细胞组织工程化软骨的保护作用并探讨其机制。方法首先将体外单层扩增的大鼠骨髓间充质干细胞在藻酸钠微球中行三维立体软骨细胞定向培养，其定向分化程度通过甲苯胺蓝染色、免疫组织化学等方法鉴定。然后将从微球释放的分化软骨细胞接种于壳聚糖．明胶复合支架，培养3周后制备组织工程化软骨，其内部软骨细胞以及支架形态用扫描电镜和激光共聚焦显微镜观察。将白细胞介素（IL）-1B作用于该细胞．支架材料上，并用白藜芦醇和（或）特异性的抗整合素81亚单位抗体进一步干预，蛋白印迹法检测Ⅱ型胶原、基质金属蛋白酶（MMP）-13以及核因子．KB的表达，扫描蛋白印迹条带灰度并进行半定量计算，采用方差分析进行统计学处理。结果在定向培养过程中，藻酸盐微球中的软骨细胞在细胞周围可形成明显的细胞外基质，促进分化，软骨细胞特异性Ⅱ型胶原和蛋白聚糖表达显著增高。定向分化后的软骨细胞在壳聚糖．明胶复合支架材料上能良好地贴壁、增殖和迁徙，形成组织工程化的软骨。蛋白印迹半定量分析：软骨Ⅱ型胶原对照组表达量0．484±0．006；白藜芦醇干预后表达量0．474±0．014，两者差异无统计学意义（P〉0．05）；IL-1β干预后，表达量下降至0．155±0．009，差异有统计学意义（P〈0．05）；而白藜芦醇能阻断IL-1β的负面效应，使Ⅱ型胶原表达量恢复至0．468±0．014，差异有统计学意义（P〈0．05），同对照组比较差异无统计学意义（P〉0．05）；而抗β1整合素能阻断白藜芦醇对IL-18的拮抗作用，使胶原表达量下降至0．169±0．011，差异有统计学意义（P〈0．05），与IL-1β单独作用比较差异无统计学意义（P〈O．05）。蛋白聚糖半定量统计和Ⅱ型胶原的表达趋势相同，同时伴随MMP-13、核因子

英文摘要：

Objective To investigate the mechanism of protective effects of resveratrol on tissue- engineered cartilage. Methods The chondregenesis of alginate-encapsulated bone marrow mesenchymal stem cells （BMSCs） were evaluated by toluidine blue staining and immunostain. The morphology of BMSCs-derived chondrocytes cultured on chitosan-gelatin scaffolds （CGS） was evaluated by scanning electron microscope and laser confocal microscope. When these cells on CGS were pre-stimulated with interleukin-l[3 （IL-1β） or co- treated with IL-1β and resveratrol in the absence and presence of specific β1-integrin blocking antibody, collagen type II , aggrecan, matrix metalloproteinase-13 （MMP-13） expression, and the translocation of nuclear factor kappaB （NF-KB） were analyzed by Western blotting. ANOVA was used for statistical analysis. Results Alginate bead culture plus conditional medium together could induce the cartilage-specific collagen type II , aggrecan expression and extracellular matrix accumulation in differentiated chondreeytes. CGS supported differentiated cell attachment, proliferation, and migration. When those cells cultured on CGS were stimulated with IL-1β alone, collagen type II and aggrecan expression was inhibited. However, MMP-13 expression increased. By Western blotting semi-quantitative analysis, the expression level of cartilage-specific collagen type II of the control group was 0.484±0.006; the expression level of resveratrol intervention group was 0.474±0.014. The difference between these two groups was not statistically significant （P〉0.05）. The expression level of the IL-1β intervention group reduced to 0.155±0.009, which was statistically significant different from the above two groups（P〈0.05）. Resveratrol could antagonist the negative effect of IL-1β, and increase collagen type II to 0.468±0.014, the difference between these two was statistically significant （P〈 0.05）, and no significant difference when compared to the control group （P〉O.05）. Specific ?