核因子相关因子-2信号通路在血红素氧合酶-1抗乙醇诱导成骨细胞损伤中的作用  

Effect of nuclear factor-E2-related factor 2 signal pathway on the heme oxygenase-1 against ethanol-induced osteoblast damage

作　　者：李杰[1] 张丰泉[2] 陈光[1] 于博凡[1] 杜振宁[1] 蔡腾[1] 

机构地区：[1]河南省人民医院急诊创伤外科,郑州450003 [2]新乡医学院公共卫生学院,新乡453003

出　　处：《中华实验外科杂志》2017年第3期440-442,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察血红素氧合酶-1（HO-1）抑制乙醇对成骨细胞损伤的作用,探讨核因子相关因子-2（Nrf-2）/抗氧化反应元件（ARE）信号通路在其保护作用中的意义.方法 提取并培养人成骨细胞,Lipofectamine 2000质粒转染HO-1,乙醇作用24h,Western bolt检测细胞中Nrf-2和HO-1蛋白的表达量,同时应用酶标仪检测成骨细胞中髓过氧化物酶（MPO）和谷胱甘肽过氧化物酶（GSH-Px）的活性.结果 阳性对照组中MPO和GSH-Px酶的活性分别为（54.17±20.61）mU/mg 蛋白和（7.94±2.42）U/mg蛋白,明显低于阴性对照组和HO-1组细胞中MPO和GSH-Px酶的活性（P=0.023、0.012）;HO-1组细胞中MPO和GSH-Px酶的活性分别为（286.54±13.82）mU/mg蛋白和（38.59±5.30）U/mg蛋白,均明显高于阳性对照组和阴性对照组细胞中MPO和GSH-Px酶的活性（P=0.000、0.000）.Western blot 结果显示,阳性对照组细胞中Nrf-2和HO-1蛋白表达量（分别为0.27±0.08和0.81±0.05）明显低于阴性对照组（分别为0.62±0.11和1.26±0.20,P=0.016、0.003）;HO-1组细胞中Nrf-2和HO-1蛋白表达量明显升高（分别为0.94±0.10和2.43±0.13,P=0.000、0.000）.结论 转染HO-1抑制乙醇诱导成骨细胞损伤的保护机制与其对细胞中Nrf-2/ARE信号通路的保护有关,其能通过Nrf-2/ARE信号通路激活并增强成骨细胞的抗氧化能力,从而减轻乙醇对成骨细胞的损伤.Objective To find the mechanism of the heme oxygenase-1 （HO-1） against ethanol-induced osteoblast damage, and clarify the effect of nuclear factor-E2-related factor 2 （Nrf-2）/antioxidant response element （ARE） signal pathway on the protective effect of HO-1, to provide theoretical support for the later research and the clinical application of biological agent of HO-1.Methods The osteoblast cells were isolated and cultured, then HO-1 was transfected using Lipofectamine 2000.After cells were exposed to ethanol for 24 h, the levels of Nrf-2 and HO-1 were measured by Western blotting.The enzyme activity of myeloperoxidase （MPO） and glutathione peroxidase （GSH-Px） were tested using the corresponding kits.Results The enzyme activity of MPO and GSH-Px was respectively （54.17±20.61） mU/mg prot and （7.94±2.42） U/mg prot in the positive group, which was significantly decreased （P=0.023, 0.012） as compared with that in the negative group.The enzyme activity of MPO and GSH-Px was significantly increased to （286.54±13.82） mU/mg prot and （38.59±5.30） U/mg prot （P=0.000, 0.000） in HO-1 group as compared with that in the negative group and positive group.The Nrf-2 and HO-1 levels （respectively 0.27±0.08 and 0.81±0.05） were significantly decreased than those in the positive group （respectively 0.62±0.11 and 1.26±0.20, P=0.016, 0.003）, but Nrf-2 and HO-1 were all significantly increased in the HO-1 group（respectively 0.94±0.10, 2.43±0.13;P=0.000, 0.000）.Conclusion The protective effect of HO-1 against ethanol-induced osteoblast damage was related to Nrf-2/ARE signal pathway, which could activate and enhance the celluar antioxidant ability to reduce the damage of the osteoblast cells induced by alcohol.

关 键 词：血红素氧合酶-1 成骨细胞 核因子相关因子-2 髓过氧化物酶 谷胱甘肽过氧化物酶 

分 类 号：R681.8[医药卫生—骨科学]