鱿鱼皮胶原蛋白水解肽对镉抑制MC3T3-E1增殖、分化及钙化的影响

Effect of Collagen Peptide Extracted from Dosidicus gigas Skin on Proliferation,Differentiation and Calcification of MC3T3-E1 Cell Induced by Cd

目的:探究秘鲁鱿鱼皮胶原蛋白水解肽增强MC3T3-E1细胞抗骨质疏松的作用。方法:将培养的MC3T3-E1细胞分为正常对照组、氯化镉损伤组和胶原蛋白水解肽干预组;利用MTT法确定氯化镉的半数抑制浓度,建立细胞损伤模型;根据各组细胞增殖率差异,确定最佳胶原蛋白水解肽的添加量;通过细胞周期、凋亡,碱性磷酸酶活性的测定及Alizarin red染色法分析胶原蛋白水解肽在细胞增殖、分化、钙化阶段拮抗氯化镉的抑制作用。结果:与氯化镉损伤组相比,胶原蛋白水解肽干预组细胞活性升高(P<0.01);处于G1期的细胞数量减小(P<0.05),S期细胞数量增大(P<0.05);凋亡率降低(P<0.05);9,12,15 d时AKP活性升高(P<0.05,P<0.01,P<0.01)。结论 :摄入一定剂量的氯化镉会抑制MC3T3-E1成骨细胞增殖、分化和钙化。胶原蛋白水解肽在一定程度上可改善此类损伤对MC3T3-E1细胞的影响。

Objective： Investigations were carried out to explore the effect of Dosidicus gigas collagen peptide on en- hancing the function of anti-osteoporosis in MC3T3-E1 cell. Method： MC3T3-E1 cells were randomly divided into three groups： the normal group, CdC12 group, and collagen peptide group; ICs0 of CdC12 was determined using MTY method and the model of cell injury was established. The optimal concentration of collagen peptidc was detected by analyzing cell proliferation rate differences. In order to analyze the effect of collagen peptide extracted from Dosidicus gigas skin on proliferation, differentiation and calcification of MC3T3-E1 cell induced by CdC12, the cell cycle, apoptosis and activity of alkaline phosphatase were measured as well as alizarin red staining were observed. Result： Compared with CdC12 group, Cell activity improved （P〈0.01）, the percentage of cell during S period increased （P〈0.05）, the rate of osteoblast apoptosis and the percentage of cell during G0-G1 period declined （P〈0.01, P〈0.05） in collagen group. At the same time, AKP activity improved in the 9th, 12th and 15th day （P〈0.05, P〈0.01, P〈0.01）. Conclusion： It showed CdC12 would cause injury and apoptosis in MC3T3-E1 cell, but collagen peptide had the ability to repair the damage.