狭鳕鱼皮胶原肽对过氧化氢诱导的人成骨细胞氧化损伤作用

目的]复制过氧化氢诱导人成骨细胞氧化损伤模型,探讨狭鳕鱼皮胶原肽保护作用机制。方法]以人成骨细胞为研究对象,将对数生长期的细胞分为6组:正常对照组、H_2O_2损伤组、胶原蛋白肽低(100μg/m L)、中(300μg/m L)、高(600μg/m L)浓度组、阳性对照组(300μg/m L维生素E),CCK-8法检测成骨细胞的存活率,酶联免疫法检测细胞培养液中SOD、MDA的含量,PCR技术检测Foxo3amRNA的表达水平,Western blot技术检测Sirt1、Foxo3a的蛋白表达水平。结果]与模型组相比,胶原肽组细胞存活率及SOD的水平升高,MDA含量下降,Foxo3amRNA及蛋白表达水平下降,Sirt1蛋白表达水平上升。结论]300μmol/L H_2O_2可成功复制成骨细胞氧化损伤模型;一定浓度的胶原蛋白肽能降低MDA的含量,提高SOD水平,并具有抗氧化损伤功能,胶原肽对成骨细胞的保护作用可能与下调Foxo3a和上调Sirt1的表达水平有关。

Protective Effects of CollagenP eptide from Pollock Sk in against H2 O2 Induced Human Osteobla sts Oxidative Damage

Objective] The aim was to investigate the protective mechanism of collagen peptide from pollock skin by establishing the H2O2 in-duced human osteoblasts oxygen damaged model.Method] Human osteoblast cells in logarithmic growth phase were divided into four groups:normal control group, H2O2 induced oxygen damaged group, collagen peptide low (100 μg/mL), medium(300 μg/mL) and high(600 μg /mL) does group, positive control group(300 μg/mL VitE).CCK-8 method was performed to determine the survival rate of osteoblasts cells.The con-tent of SOD and MDA were detected by enzymic method.The expression levels of Foxo3 mRNA was determined by PCR, Western Blot was used to detect the protein expression levels of Fox3o and Sir1t .Result] Compared with the control group, the survival rate of osteoblasts cells, the level of SOD and the expression of Sirt1 protein in model group was increased, while the content of MDA, the expression of Foxo3a mRNA and Foxo3a protein were declined significantly.Conclusion] The osteoblasts oxygen damaged model was established induced by 300 μmol/L H2O2. A certain concentration of collagen peptide could reduce the content of MDA and increase the content of SOD against oxygen damage .The mecha-nism might be related to down-regulating the expression of Foxo3a and up-regulating the expression of Sirt1.