高铁环境下成骨细胞增殖和凋亡与氧化应激的关系

The relation between osteoblast cell proliferation, apoptotic and oxidative stress under the environment of high iron

目的了解高铁环境对人成骨细胞(hFOB1.19)增殖和凋亡的影响,观察其与氧化应激的关系。方法 34℃条件下体外培养hFOB1.19,以不同浓度(50、100、200μmol/L)枸橼酸铁铵(FAC)干预48h后,用MTT法检测成骨细胞增殖活性;用流式细胞仪检测成骨细胞凋亡情况;分别用丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)试剂盒检测成骨细胞MDA含量及SOD、GSH-PX活性。结果用不同浓度FAC干预成骨细胞48h后,FAC50、100和200μmol/L干预组细胞增殖活性分别为(0.63±0.02)、(0.55±0.03)和(0.46±0.04),均显著低于对照组(0.69±0.04),呈剂量依赖性;FAC50、100和200μmol/L干预组细胞凋亡率分别为(6.98±0.84)%、(11.82±0.66)%和(15.78±0.95)%,均显著高于对照组[(3.71±0.43)%],呈剂量依赖性升高,差异有统计学意义(P<0.05);成骨细胞脂质过氧化物MDA水平呈剂量依赖性升高,成骨细胞抗氧化物酶SOD及GSH-PX活性呈剂量依赖性降低,差异有统计学意义(P<0.05)。结论高铁环境可抑制成骨细胞增殖,促进成骨凋亡,其作用机制与氧化应激有一定相关性。

Objective To study the effect of iron overload on proliferation and apoptofic in human osteoblast cells and observe its relation with oxidative stress. Methods The human osteoblast cells were cultured at 34 ℃ in vitro. The cultures were additioned with different concentrations of FAC （50 μmol/L, 100 μmol/L, 200 μmoL/L） for 48 h. Proliferation viability of osteoblasts were evaluated by MTT and apoptotic hallmarks were detected by flow cytometer; Maleic dialdehyde （MDA） content, superoxide dismutase （SOD） and glutathione peroxidase （GSH-PX） activity were measured with MDA, SOD and GSH-PX commercial kits. Results After interfered with different concentrations of FAC for48 h, the cell proliferation viability were （0. 63 ±0. 02）, （0. 55 ±0. 03） and （0. 46 ±0. 04） in the FAC 50, 100 and 200 μmol/L treatment groups, they were all significantly lower than the control group （0. 69 ± 0. 04 ） , and showed a dose-dependent manner. The cell apoptosis rates were （6. 98 ± 0. 84） % , （ 11.82 ±0. 66） % and （ 15.78 ±0.95 ） % in the FAC 50, 100 and 200 μmol/L treatment groups, they were all significantly higher than the control group （3.71 ± 0. 43 ） %, and showed a dose-dependent manner （P 〈 0. 05）. Concurrently, the lever of MDA was increased significantly with concentration-dependently, and the activity of SOD and GSH-PX were decreased significantly with concentration-dependently （P 〈 0. 05）. Conclusion Iron overload could inhibited the growth and induced the apoptotic of osteoblast, and the mechanism has a relation with oxidative stress.