纳米二氧化硅致Nrf-2基因缺陷人永生化表皮细胞氧化损伤的研究

发布时间：2018-03-21 04:25

本文选题：核因子E相关因子　切入点：纳米二氧化硅　出处：《环境与职业医学》2017年06期 　论文类型：期刊论文

【摘要】：[目的]探讨纳米二氧化硅(nano-SiO_2)致核因子E2相关因子2(Nrf-2)基因缺陷型人永生化表皮细胞的氧化应激损伤效应。[方法]采用RNA干扰技术构建人永生化表皮细胞(HaCaT)Nrf-2基因缺陷细胞模型,以终质量浓度(后称浓度)为2.5、5.0、10.0 mg/L的nano-SiO(215 nm粒径)分别染毒HaCaT细胞和Nrf-2基因缺陷细胞24 h,检测细胞内活性氧(ROS)、8-羟化脱氧鸟苷(8-oHdG)、丙二醛(MDA)、总超氧化物歧化酶(T-SOD)和谷胱甘肽过氧化物酶(GSH-Px)等氧化应激指标,并对细胞总蛋白和核蛋白中Nrf-2蛋白的表达水平进行分析。[结果]5.0、10.0 mg/L nano-SiO_2可引起HaCaT细胞的活力明显降低(P0.05),而2.5 mg/L nano-SiO_2开始即可引起Nrf-2基因缺陷细胞活力呈剂量依赖性降低(r=-0.914,P0.05),Nrf-2基因缺陷细胞的活力均明显低于同浓度组的HaCaT细胞(P0.05)。各浓度组细胞内ROS、8-oHdG和MDA水平均呈剂量依赖性增高,而T-SOD和GSH-Px的含量则呈剂量依赖性降低;与同浓度组HaCaT细胞相比,Nrf-2基因缺陷细胞中相关指标的变异程度更大,差异有统计学意义(P0.05)。正常HaCaT细胞中,低剂量组总Nrf-2蛋白和低、中剂量组核Nrf-2蛋白的表达水平增高(P0.05);而随着nano-SiO_2暴露浓度的进一步增高,两种蛋白的表达水平又呈降低的趋势。与HaCaT细胞的阴性对照比较,Nrf-2基因缺陷细胞中总Nrf-2蛋白表达水平下降,核Nrf-2蛋白表达水平未见差异。[结论]5.0 mg/L nano-SiO_2可体外诱导HaCaT细胞发生氧化应激性损伤,Nrf-2基因缺陷可通过改变细胞的抗氧化防御能力,增加HaCaT细胞对nano-SiO_2的敏感性。
[Abstract]:[Objective] to investigate the nano silica (nano-SiO_2) induced nuclear factor E2 related factor 2 (Nrf-2) gene deficient human oxidative stress damage. Methods the immortalized epidermal cells construct immortalized human epidermal cells by RNA interference (HaCaT) Nrf-2 gene deficient cell model, with the final concentration (later known as concentration) 2.5,5.0,10.0 mg/L nano-SiO (215 nm diameter) were exposed to HaCaT cells and Nrf-2 deficient cells 24 h, detection of intracellular reactive oxygen species (ROS), 8- hydroxy deoxyguanosine (8-oHdG), malondialdehyde (MDA), total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) oxide stress index, analysis. Results]5.0,10.0 mg/L nano-SiO_2 could induce HaCaT cell activity decreased and the Nrf-2 protein in total cell protein and nuclear protein expression level (P0.05), and 2.5 mg/L nano-SiO_2 can induce Nrf-2 gene. In a dose dependent decrease in cell viability (r=-0.914, P0.05), Nrf-2 gene deficient cell viability was significantly lower than that of the same concentration of HaCaT cells (P0.05). The concentration of intracellular ROS, 8-oHdG and MDA levels were dose dependently increased, while the content of T-SOD and GSH-Px showed a dose-dependent reduction; compared with the same concentration of group HaCaT cells, the variation degree of indexes related to Nrf-2 gene defects in cells is larger, the difference was statistically significant (P0.05). Normal HaCaT cells, the low dose group of total Nrf-2 protein and low expression level in middle dose group nuclear Nrf-2 protein (P0.05); and with the further increase of nano-SiO_2 concentration the expression of two proteins was decreased. Compared with the negative control of HaCaT cells, decreased levels of total Nrf-2 protein expression in Nrf-2 deficient cells, the expression of nuclear Nrf-2 protein level no difference. Conclusion:]5.0 mg/L Nan O-SiO_2 can induce oxidative stress damage in HaCaT cells in vitro, and Nrf-2 gene defects can increase the sensitivity of HaCaT cells to nano-SiO_2 by altering the antioxidant defense ability of cells.

【作者单位】：上海市疾病预防控制中心化学品毒性检定所;
【基金】：国家自然科学基金项目(编号:81302468) 上海市卫生计生委资助项目(编号:20124080) 上海市第四轮公共卫生三年行动计划“高端海外研修团队培养计划”项目(编号:GWTD2015S03)
【分类号】：R114

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