番茄红素对氧化应激后成骨细胞的影响

The effect of lycopene on osteoblasts which have incurred oxidative stress

目的研究具有强抗氧化作用的番茄红素对氧化应激后成骨细胞的影响。方法制造氧化应激状态成骨细胞模型。将细胞随机分为4组:高、中、低浓度番茄红素组及对照组。然后用流式细胞术检测各组成骨细胞生长指数,用酶标仪检测各组成骨细胞内碱性磷酸酶的活性。在各组细胞中另加入矿化结节诱导液,观察各组成骨细胞矿化能力。结果各实验组成骨细胞较对照组生长指数高,碱性磷酸酶表达增加,形成的矿化结节多。结论番茄红素可以增加氧化应激状态成骨细胞的增殖、碱性磷酸酶的表达及矿化功能。

Objective To examine the effect of lycopene-a potent antioxidant on osteoblasts which have incurred oxidative stress. Methods The crania of 10 Sprague-Dawley rats which are born within 24 hours are obtained in aseptic condition. After enzyme digestion, isolation, purified osteoblasts of SD rats are cultured for further study. The cells of the third generation are exposed to the medium containing low carbohydrates, 10^-4 mol/L H2O2 , 10% （v/v） FBS for 24 hours. Then the cells are divided into 4 groups at random：the control group,the groups treated by 10 5 mol/L,5 × 10^-6 tool/L, 10^-6 mol/L lycopene. After 72 hours of culture, the following study are continued： The proliferation index （PI） of osteoblasts in each group is determined by Flow CytoMeter;The expression of alkaline phosphatase in each group is measured by microplate reader;The cells are cultured in the medium containing 50 μg/mL vitamin C, 10 2 mol/L sodium glycerophosphate, 10% FBS and the medium is changed every 3 days. 4 weeks later,the mineral nodes of osteoblasts are dyed in alizarin red and improved Von kossa way, then counted. Results Lecopene at 10^-6 mol/L, 5 × 10^-6 mol/L, 10^-5 mol/L can increase the proliferation index （PI） of osteoblasts. The absorbency at 405 nm in the experimental groups are higher than in the control group, the absorbency in 10 5 mol/L lycopene group is the highest. Each experimental group can produce more mineral nodes than the control group. Conclusion Lecopene has an effect on stimulating the proliferation, secretion and mineralization of osteoblasts which have incurred oxidative stress.