血红素加氧酶-1基因转染对内毒素诱导的ECV304细胞氧化损伤的影响

Heme oxygenase-1 gene transfer reduces oxidative injury of ECV304 cells induced by lipopolysaccharides

目的探讨血红素加氧酶-1(HO-1)基因转染对脂多糖诱导的ECV304细胞氧化损伤的影响。方法利用逆转录病毒介导的基因转染技术将HO-1基因转染入人脐静脉内皮细胞(ECV304),应用RT-PCR和Western印迹技术检测转染细胞HO-1 mRNA和蛋白表达水平。未转染和转染HO-1基因的ECV304细胞分别培养于含或不含脂多糖(5 mg/L)的DMEM培养基24 h后,检测细胞脂质过氧化产物丙二醛(MDA)含量和乳酸脱氢酶(LDH)释放率。结果 HO-1基因和蛋白在转染的ECV304细胞的表达量显著升高。与ECV304细胞相比,转染HO-1基因的ECV304细胞MDA含量和LDH释放率均下降(P<0.05);应用HO-1抑制剂锌原卟啉共孵育后,转染细胞的MDA含量和LDH释放率增高。结论 HO-1的基因转染可增强ECV304细胞对抗脂多糖氧化损伤的能力。

Objective To investigate the effect of overexpression of heme oxygenase-1 （ HO-1 ） gene on ECV304 cells injury induced by lipopolysaccharides. Methods With gene recombination and transfer techniques, ？ HO-1 gene was transfected into human umbilical vein endothelial cells （ECV304）. Expression of HO-1 mRNA and protein was analyzed in transfected cells by RT-PCR or Western blot. Transfected cells and ECV304 cells were cuhured in DMEM media only or with lipopolyssachades （5 mg/L） for 24 h. Lipid peroxidation in cells was tested as malondialdehyde content. The release rate of lactate dehydrogenase was measured by colorimetric assay. Results The expression of HO-1 mRNA and protein were increased in transfected cells. Treatment of ECV304 cells with lipopolyssachades caused an increase in malondialdehyde contents and lactate dehydrogenase release rate, while transfeeted cells showed lower malondialdehyde contents and lactate dehydrogenase release rate compared to those untransfected cells with the same treatment. The protection effect of HO-1 was abolished when HO-1 inhibitor zinc protoporphyrin-IX was used in transfected ceils. Conclusions The upregulation of HO-1 gene expression was able to enhance endothelial cell resistance against oxidative injury induced by lioooolysaccharides.