葛根素对过氧化氢致成骨细胞氧化损伤的影响

The effect of puerarin on the damage of osteoblasts induced by hydrogen peroxide

目的研究具有抗氧化作用的植物雌激素——葛根素(puerarin,Pue)对氧化应激后成骨细胞(osteoblast,OB)的影响。方法采用酶多次消化法,由出生24 h以内的Wistar大鼠头盖骨分离OB,培养、传代及诱导分化、矿化等。用0.1 mmol/L H_2O_2制造氧化应激状态模型。将细胞随机分为对照组、H_2O_2组[H'组]、"Pue(10^(-10)mol/L^10^(-3)mol/L)+H_2O_2"组["P+H'"组]和"H_2O_2+Pue(10^(-10)mol/L^10^(-3)mol/L)"组["H'+P"组]。观察不同处理组细胞的增殖、分化功能,并测定其微量丙二醛(m MDA)含量、超氧化物歧化酶(super oxide dismutase,SOD)活性、总抗氧化能力(T-AOC)等。结果与H_2O_2组相比,"P+H'"组和"H'+P"组中Pue10^(-7)mol/L^10^(-4)mol/L使细胞的增殖、分化功能,SOD活性和T-AOC均有显著升高(P<0.01),而mMDA含量则显著降低(P<0.01)。结论 H_2O_2过量会导致OB的氧化损伤,而葛根素在体外对H_2O_2介导OB的氧化损伤具有保护作用。

Objective To investigate the effect of a potent anti-oxidative phytoestrogen,puerarin,on osteoblasts under oxidative stress.Methods Osteoblasts were isolated from the calvarias of new-born Wistar rats with enzyme digestion method and cultured.The cells of the first to third generation were randomly divided into 4 groups：the control group,the 10^（-4） mol/L H_2O_2 group,the 10^（-4） mol/L H_2O_2＋10-5 mol/L puerarin group,and the 10^（-5） mol/L puerarin group.The proliferation and differentiation of osteoblasts were determined.The content of maleic dialdehyde（MDA）,the activity of superoxide dismutase（SOD）,and total anti-oxidative capability（T-AOC） were tested and compared.Results Compared with those in the H_2O_2 group,the proliferation and differentiation of osteoblasts,the activity of SOD,and T-AOC in the model groups increased significantly（P〈0.01）.However,the content of MDA decreased significantly（P〈0.01）.Conclusion Over-dose of H_2O_2 can cause oxidative damage in osteoblasts.Puerarin can prevent osteoblasts from oxidative damage in vitro.