水解鱼胶原诱导大鼠骨髓间充质干细胞成骨分化的作用

Effect of hydrolyzed fish collagen on the osteogenic differentiation of rat bone marrow mesenchymal stem cells

目的探讨水解鱼胶原诱导大鼠骨髓间充质干细胞(rBMSCs)成骨分化的潜能。方法制备获得水解鱼胶原,对其分子量,氨基酸成分和接触角进行表征,利用MTT试验和Real-Time PCR试验分别研究水解鱼胶原对rBMSCs细胞活力及成骨分化相关基因ALP,OCN和RUNX2的影响,利用western blot技术探讨水解鱼胶原对ERK1/2信号通路的影响。结果水解鱼胶原的分子量为700-1300 Da,接触角约为26度,主要含甘氨酸,脯氨酸和羟脯氨酸。水解鱼胶原能提高rBMSCs细胞的活力,促进成骨分化相关基因ALP,OCN和RUNX2的表达,Western Blot结果显示水解鱼胶原可激活ERK1/2信号通路,进而促进RUNX2蛋白表达上调。结论水解鱼胶原具有诱导大鼠骨髓间充质干细胞成骨分化的潜能。

Objective The aims of this study were to investigate the influence of hydrolyzed fish collagen（HFC） on osteogenic differentiation of rat bone marrow mesenchymal stem cells（rBMSCs）. Methods In this study, the hydrolyzed fish collagen were extracted from the fish scale. The molecular weight, amino acid composition, and contact angle of HFC were measured. The influence of HFC on cell viability were assessed by MTT, the expression of osteogenic genes ALP, OCN and RUNX2 was investigated by Real Time-PCR. Western blotting were used to examine the role of ERK1/2 signaling pathway. Results The molecular weight of HFC ranged from 700 to 1300 Da, the contact angle of HFC was approximately 26°, and HFC was found to be composed of various amino acids, including glycine, proline, and hydroxyproline. The results showed that HFC was favorable for cell growth. The expression of the osteogenic gene marker, ALP, OCN, and RUNX2 was enhanced by HFC treatment. HFC could activate the ERK1/2 signaling pathway and then increase the protein level of RUNX2. ConclusionHFC has the ability to actively promote osteogenic differentiation of rBMSCs.