生长激素和细胞因子对兔关节软骨细胞增殖和基质分泌的影响

Effect of growth hormone and cytokine on cellular proliferation and matrix secretion of articular chondrocytes in rabbits

目的:探索生长激素对体外培养的关节软骨细胞的影响及其与细胞因子的关系,为生长激素在软骨组织工程中的应用提供依据。方法:实验于2003-11/2004-03在南京医科大学分子生物研究所完成。消化获取3个月龄新西兰兔关节软骨细胞,随机分为胰岛素样生长因子Ⅰ、转化生长因子β1、生长激素、生长激素+胰岛素样生长因子Ⅰ、无因子对照组进行培养,每组16孔细胞。以噻唑蓝法检测各组细胞增殖率、阿新蓝染色法检测各组细胞聚胺多糖分泌量。结果:①各处理组细胞增殖率均高于对照组(P<0.05),生长激素和细胞因子均能促进软骨细胞增殖,其中胰岛素样生长因子Ⅰ+生长激素组细胞增殖最显著,是对照组的2.17倍[噻唑蓝法检测不同组吸光度:无因子对照组为0.199±0.05,转化生长因子β1组为0.317±0.067,胰岛素样生长因子Ⅰ组为0.384±0.084,生长激素组为0.252±0.056,生长激素+胰岛素样生长因子Ⅰ组为0.433±0.080]。②各处理组培养上清聚胺多糖含量均高于对照组(P<0.05),生长激素和细胞因子均能促进软骨细胞基质分泌,其中胰岛素样生长因子Ⅰ+生长激素组聚胺多糖含量最高,是对照组的2.10倍[阿新兰染色法检测不同因子组上清聚胺多糖含量:无因子对照组为(49.69±1.37)mg/L,转化生长因子β1组为(80.10±1.42)mg/L,胰岛素样生长因子Ⅰ组为(88.44±1.68)mg/L,生长激素组为(67.73±1.56)mg/L,生长激素+胰岛素样生长因子Ⅰ组为(104.47±1.86)mg/L]。结论:生长激素也有促进体外培养的关节软骨细胞增殖和基质合成作用,胰岛素样生长因子Ⅰ与生长激素联用有协同作用。

AIM： To probe into the effect of growth hormone（GH） on articular chondrocyte cultured in vitro and the correlation with cytokine, providing basis for the application of GH in cartilage tissue engineering. METHODS： This experiment was conducted at the Institute of Molecular Biology , Nanjing Medical University from November 2003 to March 2004. The primary articular chondrocytes of 3-month-old rabbit were harvested and randomly divided into insulin growth factor-1 （IGF-1）, transforming growth factor-β1（TGF-β1 ）,GH , combination of 1GF-1 and GH and control group （no factor）, 16 wells in each group. Then cells were cultured with corresponding factors . The cellular proliferative rate was detected with methylthiazolyl tetrazolium （MTI）assay in each group. Glycosaminoglycans （GAG） was determined by Alcian Blue staining. RESULTS： ①The proliferative rate of chondrocytes cultured with IGF-1, TGF-β1 , GH, combination of IGF-1 and GH were significantly higher than those of control group（P 〈 0.05 ） . GH and cytokines both ccould promote the proliferation of chondrocytes. Cellular proliferation in the IGF-1 ＋ GH was the most significantly, which was 2.17 times of control group [The absertance detected by MTT assay： control group 0199 ±0.05,TGF-β1group 0.317±0.067,IGF-1 group 0.384±0.084,GH group 0.252±0.056,GH＋IGF-1 group 0.433±0.080]. ②The content of GAG in each treated group was significantly higher than that of control group （P 〈 0.05）. GH and cytokines both could promote the matrix secretion of chondrocytes.The combination group got the highest concentration of GAG ,which was 2.10 times of control group[control group （49.69±1.37） mg/L ,TGF-β1 group （80.10±1.42） mg/L, IGF-1 group （88.44±1.68） mg/L, GH group （67.73±1.56） mg/L, GH＋IGF-1 group （104.47±1.86） mg/L]. CONCLUSION： GH can also promote the proliferation and matrix synthesis of articular chondrocytes cultured in vitro, and it has synergistic action with IGF-1.