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[慢性阻塞性肺疾病患者EPHX1基因多态性与抗氧化能力的关系研究]

[Study on the relationship between EPHX1 gene polymorphism and antioxidant capacity in patients with chronic obstructive pulmonary disease].

作者信息

Luo Y, Dai L M, Jia M, Zhao Z H, Hu C M, Qi W Y, Zhang J Q

机构信息

Second Department of Respiratory and Critical Care Medicine, First Affiliated Hospital of Kunming Medical University, Kunming 650032, China.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2019 Oct 12;42(10):760-764. doi: 10.3760/cma.j.issn.1001-0939.2019.10.009.

Abstract

To explore the difference of mRNA, protein expression levels and the indexes of peripheral blood antioxidant capacity in peripheral blood lymphocytes of different EPHX1 genotypes in chronic obstructive pulmonary disease(COPD). A case-control study was conducted to collect peripheral blood samples of 220 stable chronic COPD patients with smoking history and 230 healthy smokers (control group) from October 2016 to February 2018 in the First Affiliated Hospital of Kunming Medical University, and the genetic testing was carried out according to the operation instructions of BigDye Terminator v1.1 DNA Sequencing Kit. Based on their EPHX1 exon 3 and exon 4 polymorphism status, the EPHX1 was classified into 4 groups, i. e., normal activity, slow activity, extremely slow activity and fast activity. Then COPD patients were allocated to either a slow activity group (slow and very slow activity) or a fast activity group (normal and fast activity) according to EPHX1 genotype and gene activity. The expression of EPHX1 mRNA and protein in peripheral blood lymphocytes were detected by qRT-PCR and Western blot, and indexes of serum antioxidant capacity was detected by corresponding kits. (1)The 2(-ΔΔCt) of the control group was 1.000, and the 2(-ΔΔCt) of the COPD group was 1.052±0.023. There was no significant difference in the level of EPHX1 mRNA expression between the two groups (1.992 0.865). The level of EPHX1 mRNA expression in the slow activity group was not different significantly compared to that in the fast-active group (1.053±0.023 vs 1.048±0.021, 1.133, 0.260). (2)The level of EPHX1 protein expression by Western blot analysis showed that the EHPX1/GAPDH gray ratio was not different significantly between the COPD group and the control group (0.613±0.089 vs 0.602±0.075, 0.805, 0.422). The level of EPHX1 protein expression in the slow activity group was not significantly different compared to that in the fast activity group (0.606±0.088 vs 0.622±0.092, -0.786 0.434). (3)There were significant differences in indexes of antioxidant capacity between the control group and the COPD group (0.05). There were significant differences in indexes of antioxidant capacity between the slow activity group and the fast activity group of COPD patients (0.05). The different antioxidant capacity of COPD patients with different EPHX1 genotypes may be related to the polymorphism of EPHX1 gene affecting the activity of microsomal epoxidase, but not to the level of EPHX1 mRNA and protein expression.

摘要

探讨慢性阻塞性肺疾病(COPD)不同EPHX1基因型患者外周血淋巴细胞中mRNA、蛋白表达水平及外周血抗氧化能力指标的差异。采用病例对照研究方法,于2016年10月至2018年2月在昆明医科大学第一附属医院收集220例有吸烟史的稳定期慢性COPD患者及230例健康吸烟者(对照组)的外周血样本,并按照BigDye Terminator v1.1 DNA测序试剂盒操作说明进行基因检测。根据EPHX1外显子3和外显子4多态性状态,将EPHX1分为正常活性、慢活性、极慢活性和快活性4组。然后根据EPHX1基因型和基因活性将COPD患者分为慢活性组(慢活性和极慢活性)和快活性组(正常活性和快活性)。采用qRT-PCR和Western blot检测外周血淋巴细胞中EPHX1 mRNA和蛋白的表达,并用相应试剂盒检测血清抗氧化能力指标。(1)对照组2(-ΔΔCt)为1.000,COPD组为1.052±0.023。两组间EPHX1 mRNA表达水平差异无统计学意义(1.992 0.865)。慢活性组EPHX1 mRNA表达水平与快活性组比较差异无统计学意义(1.053±0.023 vs 1.048±0.021,1.133,0.260)。(2)Western blot分析显示,COPD组与对照组EHPX1/GAPDH灰度比值差异无统计学意义(0.613±0.089 vs 0.602±0.075,0.805,0.422)。慢活性组EPHX1蛋白表达水平与快活性组比较差异无统计学意义(0.606±0.088 vs 0.622±0.092,-0.786 0.434)。(3)对照组与COPD组抗氧化能力指标差异有统计学意义(0.05)。COPD患者慢活性组与快活性组抗氧化能力指标差异有统计学意义(0.05)。不同EPHX1基因型COPD患者抗氧化能力不同,可能与EPHX1基因多态性影响微粒体环氧化酶活性有关,而与EPHX1 mRNA和蛋白表达水平无关。

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