Abstract：To investigate the DNA damage from oxidative stress in human umbilical vein endothelial cells (hUVECs) induced by cholesterol, the γH2AX foci were detected by immunofluorescence following cholesterol treatments at 125 mg/L, 25 mg/L, 50 mg/L for 12 hours. The results showed that formation of γH2AX foci was increased as measured by the numbers fluorescence intensity. Western bolt and flow cytometry (FCM) showed that the level of γH2AX and reactive oxygen species (ROS) increased with cholesterol doses. Using comet assay to detect DNA lesions, we found higher percentage of tailing cells as the cholesterol concentration increased. Preexposed to 10 mmol/L Nacetyl6cysteine(NAC) for 1 hour, the hUVECs treated with 50 mg/L cholesterol showed significantly reduced γH2AX foci and decreased ROS. The results indicated that cholesterol induced DNA damage in hUVECs by elevated ROS levels.
汪慧星, 杨一峻, 钱民章. 胆固醇致人血管内皮细胞DNA损伤[J]. 中国生物化学与分子生物学报, 2012, 28(5): 442-448.
WANG Hui-Xing, YANG Yi-Jun, QIAN Min-Zhang. Cholesterol Induces DNA Damage in HUVECs. Chinese Journal of Biochemistry and Molecular Biol, 2012, 28(5): 442-448.
Moghadasian M H. Experimental atherosclerosis: a historical overview[J]. Life Sci, 2002, 70(8):855-865
王克勤.脂蛋白与动脉粥样硬化[M]. 北京:人民卫生出版社(Wang K Q. Lipoprotein and atherosclerosis [M]. Beijing: People’s Medical Publishing House), 1995: 417-419
Botto N, Rizza A, Colombo M G, et al. Evidence for DNA damage in patients with coronary artery disease [J]. Mutat Res, 2001, 493(1-2): 23-30
Rojas A, Figueroa H, Re L, et al. Oxidative stress at the vascular wall mechanistic and pharmacological aspects [J]. Arch Med Res, 2006, 37(4): 436-448
Mercer J R, Cheng KK, Figg N, et al. DNA damage links mitochondrial dysfunction to atherosclerosis and the metabolic syndrome[J]. Circ Res, 2010, 107(8): 1021-1031
Song Y, Leonard S W, Traber M G, et al. Zinc deficiency affect DNA damage, oxidative stress, antioxidant defense, and DNA repair in rats [J]. J Nutr, 2009, 139(9): 1629-1631
权媛,钱民章.胆固醇通过 NADPH氧化酶诱导 ROS升高,NF-κB活化进而导致内皮细胞损伤[J].中国病理生理杂志(Quan Y, Qian M Z. Cholesterol induces endothelial cells injury by increasing production of reactive oxygen species and activating NF-κB[J].Chin J Pathophysiol),2010,26(8):1521- 1526
Mahmoudi M, Mercer J, Bennett M. DNA damage And repair in atherosclerosis [J].Circ Res,2006, 7(1): 259-268
Mercer J R, Cheng K K, Figg N, et a1. DNA damage links mitochondrial dysfunction to atherosclerosis and the metabolic syndrome [J]. Circ Res, 2010, 107(8): 1021-1031
Mahley R W. The role of dietary fat and cholesterol in atherosclerosis and lipoprotein metabolism[J]. West J Med, 1981, 134(1): 34-42
Banáth J P, Klokov D, MacPhail S H, et al. Residual gammaH2AX foci as an indication of lethal DNA lesions[J]. BMC Cancer, 2010, 10: 4
Kinner A, Wu W, Staudt C, et al. γH2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin [J]. Nucleic Acids Res, 2008, 17(36):5678-5694
Wang H, Wang M, Wang H, et al. Complex H2AX phosphorylation pattern by multiple kinases including ATM and DNA-PK in human cells exposed to ionizing radiation and treated with kinase inhibitors[J]. Cell Physiol, 2005, 202(2): 492-502
Riches L C, Lynch A M, Gooderham N J. Early events in the mammalian response to DNA double-strand breaks[J]. Mutagenesis, 2008, 23(5): 331-339
Rogakou E P, Pilch D R, Orr A H, et a1. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139[J]. J Biol Chem, 1998, 273(10): 5858-5868
Rothkamm K, Lobrich M. Evidence for a lack of DNA double-strand break repair inhuman ceils exposed to very low X-rays doses[J]. Proc Natl Acad Sci U S A, 2003, l00(9): 5057-5062
Rothkamm K, Horn S. γH2AX as protein biomarker for radiation exposure[J]. Ann Ist Super Sanita, 2009, 45(3): 265-271
田梅, 潘艳, 阮健磊, 等. 60Coγ-射线诱导人膀胱癌 EJ细胞中磷酸化组蛋白 H2AX焦点形成[J]. 中国职业医学(Tian M, Pan Y, Ruan J L, et al. Phosphrylation histone H2AX foci formation in human bladder cancer EJ cells induced by 60Co γ-ray irradiation[J]. China Occup Med), 2008, 35 (3): 187-190
Klaude M, Eriksson S, Nygren J, et al. The comet assay: mechanisms and technical considerations[J]. Mutat Res, 1996, 363(2): 89-96
