中国组织工程研究2017，Vol.21Issue(4)：505-511,7.DOI:10.3969/j.issn.2095-4344.2017.04.003
低氧对H2O2预处理MC3T3-E1增殖和成骨分化功能的影响
Hypoxia effects on the proliferation and differention of hydrogen peroxide-pretreated MC3T3-E1
摘要
Abstract
BACKGROUND:The intracel ular accumulation of reactive oxygen species leads to oxidative stress. Hypoxia is widespread in physiological and pathological condition. Variation of bone proliferation and differentiation when bone tissues cultured or bone cel s induced toxicity by reactive oxygen species under hypoxia have not yet been reported. OBJECTIVE:To observe the biological characteristics of MC3T3-E1 pretreated with different concentrations of hydrogen peroxide (H2O2) in hypoxia, thus understanding the cel mechanism underlying prolonged bone healing in the elderly with osteoporosis and diabetes. METHODS:The MC3T3-E1 cel s pretreated with different concentrations of H2O2 were cultured in different oxygen concentrations. The proliferation of MC3T3-E1 was detected by cel counting kit-8. The cel differentiation was detected through alkaline phosphatase staining and alizarin red staining. Total RNAs were extracted and used for analyzing the mRNA levels of col age type 1, alkaline phosphatase and Cbfa1. RESULTS AND CONCLUSION:When MC3T3-E1 pretreated with 200μmol/L H2O2 for 6 hours, the cel proliferation was increased with time, but lower than that in the control group. The alkaline phosphatase activity was weakened, and the number of mineralized nodes was decreased at the early stage of differentiation. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours, the cel proliferation was decreased obviously. The alkaline phosphatase activity was stil weakened, and the number of mineralized nodes was decreased further, but not affected by hypoxia. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours and then cultured in hypoxia, the mRNA expression of Cbfa1 was decreased, but the mRNA expressions of col age type 1 and alkaline phosphatase were significantly increased. These results suggest that MC3T3-E1 pretreated with low concentration of H2O2 show a significant decrease in proliferation, while MC3T3-E1 pretreated with a high concentration of H2O2 and cultured in hypoxia show a decrease in osteogenic differentiation, especial y at the early stage of alkaline phosphatase formation.关键词
组织构建/成骨细胞/缺氧诱导因子/MC3T3-E1/H2O2/碱性磷酸酶/Ⅰ型胶原/cbfa1/定量PCR/国家自然科学基金分类
医药卫生引用本文复制引用
梁静,王君,唐传玲,周琦,魏立,胡方琼,万荣..低氧对H2O2预处理MC3T3-E1增殖和成骨分化功能的影响[J].中国组织工程研究,2017,21(4):505-511,7.基金项目
国家自然科学基金资助项目(81300505) （81300505）
