外源性转化生长因子-β_1作用下胎儿软骨终板细胞Ⅱ型胶原与蛋白多糖的相关性

The correlation between collagen Ⅱ and proteoglycan of the fetal endplate cartilage cells cultured by the transforming growth factor-β_1 in vitro

目的观察外源性转化生长因子-β_1(TGF-β_1)作用下胎儿软骨终板细胞Ⅱ型胶原和蛋白多糖的变化及相关性。方法用含10μg/L外源性TGF-β_1培养基培养第2代胎儿软骨终板细胞,分别培养0d、1d、2d、3d、4d、5d、6d。Western blot和RT-PCR检测培养细胞Ⅱ型胶原和蛋白多糖及其mRNA的表达。结果 0天与第1天之间相比较,第4天、第5天与第6天之间相互比较,Ⅱ型胶原和蛋白多糖及其基因表达的差异皆无统计学意义(P>0.05);第1天、第2天、第3天和第4天之间相互比较,Ⅱ型胶原和蛋白多糖及其基因表达的差异有统计学意义(P<0.05)。Ⅱ型胶原和蛋白多糖的mRNA表达及蛋白表达均呈正相关(分别为R=0.9991和R=0.9931)。结论 TGF-β_1能促进体外培养的胎儿软骨终板细胞合成Ⅱ型胶原和蛋白多糖,且二者在变化过程中具有显著的相关性。

Objective To observe the correlation and the change of collagenⅡand proteoglycan of the fetal endplate cartilage cells cultured by the transforming growth factor-β_1（TGF-β_1 ） in vitro.Methods The second generation fetal endplate cartilage cells for 0,1,2,3,4,5 or 6 days with TGF-β_1（10μg/L） in vitro.The proteoglycan,collagenⅡgene and protein expression were detected by Western blot or RT-PCR.Results There was not significant difference of the expression of collagen Hand proteoglycan mRNA before 1d and after 4d（P 0.05）,however,the expression of collagenⅡand proteoglycan mRNA increased gradually with a significant difference from Id to 4d（P 0.05）,and collagen II change with proteoglycan mRNA and expression of protein were positively correlated（R =0.9991,R =0.9931 respectively）.Conclusion The production of collagen II and proteoglycan of the cultured fetal endplate cartilage cells were promoted by TGF-β_1 in vitro,and there were significant correlation between collagenⅡand proteoglycan.