鹅骨胶原蛋白及肽对大鼠骨髓间充质干细胞增殖及向成骨分化的影响

Effects of Goose Bone Collagen and Peptides on Bone Marrow Mesenchymal Stem Cell Proliferation and Differentiation into Osteoblasts

本实验通过研究鹅骨胶原蛋白、含钙胶原蛋白、胶原肽、含钙胶原肽对大鼠骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)增殖及向成骨细胞(Osteoblast,OB)分化的影响,阐述鹅骨胶原蛋白及多肽对骨质疏松的作用机制。采用MTT(噻唑蓝)法测定鹅骨胶原蛋白及多肽对BMSCs增殖的影响;碱性磷酸酶(Alkaline phosphatase,ALP)染色、ALP活性测定、茜素红染色和骨钙素(BGP)分泌量测定分析BMSCs成骨分化能力。结果表明,相较于对照组,试验组可促进BMSCs的增殖,第14d ALP染色试验组ALP染色阳性率显著高于对照组(p<0.05),第7 d和第14 d鹅骨胶原蛋白及肽可显著提高ALP的活性表达(p<0.05),第21 d茜素红染色试验组钙结节个数相较于对照组显著增加(p<0.05),骨钙素分泌量试验组显著高于对照组(p<0.05)。由此表明,大分子胶原蛋白、含钙大分子胶原蛋白、小分子胶原多肽、含钙小分子胶原多肽均可促进BMSCs的增殖及向成骨细胞分化,以含钙小分子胶原多肽作用效果最佳。

The effects of collagen, collagen containing calcium, collagen peptides, and collagen peptides containing calcium extracted from goose bones on the proliferation of rat bone marrow mesenchymal stem cells(BMSCs) and their differentiation into osteoblasts(OBs) were studied, and the mechanism by which collagen and peptides influence osteoporosis was described. The effects of goose bone collagen and peptides on BMSC proliferation were determined by an MTT(3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. The osteogenic differentiation capacity of BMSCs was measured by alkaline phosphatase(ALP) staining, ALP activity, Alizarin Red staining, and osteocalcin secretion. The experimental groups exhibited greater proliferation of BMSCs than the control group. The positive ALP staining rate of the experimental group on day 14 was significantly higher that of the control group(p < 0.05); ALP activity levels of the experimental group on days 7 and 14 were significantly higher than that of the control group(p < 0.05). On day 21, the number of calcified nodules of the experimental groups, detected by Alizarin Red staining, was significantly high than that of the control group(p < 0.05). The osteocalcin secretion of experimental groups was significantly higher than that of the control group. These results indicated that macromolecular collagen, macromolecular collagen containing calcium, small collagen peptides, and small collagen peptides containing calcium promote BMSC proliferation and the differentiation of BMSCs into OBs, and the small collagen peptides containing calcium displayed the strongest effect.