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[细颗粒物2.5对慢性阻塞性肺疾病小鼠核因子E2相关因子2水平的影响及其与氧化应激的关系]

[Effect of PM2.5 on the level of nuclear factor erythroid-2 related factor 2 in chronic obstructive pulmonary disease mice and its relationship with oxidative stress].

作者信息

Zhao Q J, Liu X J, Zeng X L, Bao H R

机构信息

Department of Gerontal Respiratory Medicine, the First Hospital of Lanzhou University, Lanzhou 730000, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2016 Jul 26;96(28):2241-5. doi: 10.3760/cma.j.issn.0376-2491.2016.28.009.

Abstract

OBJECTIVE

To explore the effects of fine particulate matter on the level of nuclear factor erythroid-2 related factor 2 (Nrf2) in pulmonary tissues of chronic obstructive pulmonary disease (COPD) mouse models and its relationship with oxidative stress.

METHODS

Totally 40 BALB/c mice were randomly divided into normal control group, normal PM2.5 group, COPD control group and COPD PM2.5 group.COPD mice were established using exposure of cigarette smoking.PM2.5 (20 mg/kg) was intratracheally instilled in PM2.5 group mice.Mice pulmonary function was measured by mice noninvasive body plethysmograph and lung histopathology was observed in normal control group and normal PM2.5 group mice.The mRNA and protein expression of Nrf2 was measured with real-time polymerase chain reaction (PCR) and Western blot methods.Total antioxidative capacity (TAC) was measured by O-phenanthroline colorimetry.Glutathione peroxidase (GSH-PX) was measured by improved Hafeman colorimetry and malondialdehyde (MDA) by thiobarbiturieacid colorimetry.

RESULTS

Nrf2 mRNA and protein in normal control group, normal PM2.5 group, COPD control group and COPD PM2.5 group were 1.00, 4.46±0.42, 4.93±0.63, 6.41±0.35 and 0.92±0.08, 1.23±0.07, 1.20±0.09, 1.43±0.10.Nrf2 mRNA and protein in COPD control group were increased than those in normal control group while those in normal PM2.5 group and COPD PM2.5 group were respectively higher than each control group.Comparing to normal PM2.5 group, the Nrf2 mRNA and protein in COPD PM2.5 group were increased (all P<0.01). TAC and GSH-PX in each group were (5.1±0.4), (2.9±0.4), (3.3±0.3), (1.8±0.3) and (13.4±0.5), (9.9±0.7), (9.8±0.7), (7.0±0.6) U/mgpro.TAC and GSH-PX in COPD control group were decreased than those in normal control group while those in normal PM2.5 group and COPD PM2.5 group were respectively lower than each control group.Comparing to normal PM2.5 group, the Nrf2 mRNA and protein in COPD PM2.5 group were decreased (all P<0.01). MDA in each group were (2.9±0.4), (4.8±0.5), (4.5±0.3), and (6.2±0.4) nmol/mgpro.MDA in COPD control group were increased than those in normal control group while those in normal PM2.5 group and COPD PM2.5 group were respectively higher than each control group.Comparing to normal PM2.5 group, the MDA in COPD PM2.5 group were increased (all P<0.01). Positive correlations were observed between Nrf2 mRNA, protein and MDA, while negative correlations were observed between Nrf2 mRNA, protein and TAC, GSH-PX in all groups (all P<0.05).

CONCLUSIONS

PM2.5 can induce Nrf2 expression and aggravate oxidative stress in COPD mice.The increased expression of Nrf2 is closely associated with oxidative stress.

摘要

目的

探讨细颗粒物对慢性阻塞性肺疾病(COPD)小鼠模型肺组织中核因子红细胞2相关因子2(Nrf2)水平的影响及其与氧化应激的关系。

方法

将40只BALB/c小鼠随机分为正常对照组、正常PM2.5组、COPD对照组和COPD PM2.5组。采用香烟烟雾暴露建立COPD小鼠模型。对PM2.5组小鼠气管内滴注PM2.5(20mg/kg)。用小鼠无创体容积描记仪测定小鼠肺功能,并观察正常对照组和正常PM2.5组小鼠的肺组织病理学变化。采用实时聚合酶链反应(PCR)和蛋白质免疫印迹法检测Nrf2的mRNA和蛋白表达。用邻菲啰啉比色法测定总抗氧化能力(TAC)。用改良哈夫曼比色法测定谷胱甘肽过氧化物酶(GSH-PX),用硫代巴比妥酸比色法测定丙二醛(MDA)。

结果

正常对照组、正常PM2.5组、COPD对照组和COPD PM2.5组的Nrf2 mRNA分别为1.00、4.46±0.42、4.93±0.63、6.41±0.35,蛋白分别为0.92±0.08、1.23±0.07、1.20±0.09、1.43±0.10。COPD对照组的Nrf2 mRNA和蛋白高于正常对照组,正常PM2.5组和COPD PM2.5组的Nrf2 mRNA和蛋白分别高于各自对照组。与正常PM2.5组比较,COPD PM2.5组的Nrf2 mRNA和蛋白升高(均P<0.01)。各组的TAC和GSH-PX分别为(5.1±0.4)、(2.9±0.4)、(3.3±0.3)、(1.8±0.3)U/mgpro和(13.4±0.5)、(9.9±0.7)、(9.8±0.7)、(7.0±0.6)U/mgpro。COPD对照组的TAC和GSH-PX低于正常对照组,正常PM2.5组和COPD PM2.5组的TAC和GSH-PX分别低于各自对照组。与正常PM2.5组比较,COPD PM2.5组的TAC和GSH-PX降低(均P<0.01)。各组的MDA分别为(2.9±0.4)、(4.8±0.5)、(4.5±0.3)、(6.2±0.4)nmol/mgpro。COPD对照组的MDA高于正常对照组,正常PM2.5组和COPD PM2.5组的MDA分别高于各自对照组。与正常PM2.5组比较,COPD PM2.5组的MDA升高(均P<0.01)。各组中Nrf2 mRNA、蛋白与MDA呈正相关,与TAC、GSH-PX呈负相关(均P<0.05)。

结论

PM2.5可诱导COPD小鼠Nrf2表达并加重氧化应激。Nrf2表达增加与氧化应激密切相关。

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