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接受Goeckerman疗法治疗的银屑病患者中GSTM1缺失多态性的液滴数字PCR分析

Droplet Digital PCR Analysis of GSTM1 Deletion Polymorphism in Psoriatic Subjects Treated with Goeckerman Therapy.

作者信息

Beránek Martin, Fiala Zdeněk, Kremláček Jan, Andrýs Ctirad, Hamáková Květoslava, Palička Vladimír, Borská Lenka

机构信息

Department of Biochemical Sciences, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Hradec Králové, Czech Republic.

Institute of Clinical Biochemistry and Diagnostics, Charles University Hospital and Faculty of Medicine in Hradec Králové, Hradec Králové, Czech Republic.

出版信息

Acta Medica (Hradec Kralove). 2016;59(3):75-78. doi: 10.14712/18059694.2016.94. Epub 2016 Aug 29.

Abstract

Goeckerman therapy (GT) represents an effective treatment of psoriasis including a combination of pharmaceutical grade crude coal tar (CCT) and ultraviolet irradiation (UV-R). Coal tar contains a mixture of polycyclic aromatic hydrocarbons. The best known carcinogenic polyaromate - benzo[a]pyrene is metabolized into a highly reactive benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE). Glutathione S-transferase M1 (GSTM1) catalyses the conjugation of drugs, toxins and products of oxidative stress with glutathione. The aim of the study is to found possible associations between GSTM1 genotypes and the level of BPDE-DNA adducts in 46 psoriatic patients treated with GT. For genotyping, droplet digital PCR was applied. The GSTM1 copy number was normalized to β-globin reference gene. In five GSTM1*1/1 subjects, the GSTM1 to β-globin ratio moved from 0.99 to 1.03 with a median of 1.01. GSTM10/1 heterozygotes (n = 20) contained only one GSTM1 function allele which conditioned the ratio 0.47-0.53 (median 0.50). GSTM10/0 individuals (n = 21) showed no amplification of the null variants because of the large deletion in GSTM1. BPDE-DNA concentrations ranged from 1.8 to 66.3 ng/µg with a median of 12.3 ng/µg. GSTM10/0 and GSTM10/1 genotypes showed non-significantly higher concentrations of BPDE-DNA adducts than the GSTM11/*1 one (12.3 and 12.4 vs 7.8 ng/µg). The non-significant relationship between BPDE-DNA adducts and GSTM1 genotypes in psoriatic patients could be associated with relatively low doses of CCT and short-term UV-R exposures used in GT.

摘要

格克曼疗法(GT)是一种治疗银屑病的有效方法,包括药用级粗煤焦油(CCT)与紫外线照射(UV-R)联合使用。煤焦油含有多环芳烃混合物。最著名的致癌多环芳烃——苯并[a]芘会代谢为高反应性的苯并[a]芘-7,8-二醇-9,10-环氧化物(BPDE)。谷胱甘肽S-转移酶M1(GSTM1)催化药物、毒素和氧化应激产物与谷胱甘肽的结合。本研究的目的是在46例接受GT治疗的银屑病患者中寻找GSTM1基因型与BPDE-DNA加合物水平之间可能存在的关联。采用液滴数字PCR进行基因分型。将GSTM1拷贝数标准化为β-珠蛋白参考基因。在5名GSTM1*1/1受试者中,GSTM1与β-珠蛋白的比值从0.99变为1.03,中位数为1.01。GSTM10/1杂合子(n = 20)仅含有一个GSTM1功能等位基因,这使得比值为0.47 - 0.53(中位数0.50)。GSTM10/0个体(n = 21)由于GSTM1中的大片段缺失,未显示无效变异的扩增。BPDE-DNA浓度范围为1.8至66.3 ng/μg,中位数为12.3 ng/μg。GSTM10/0和GSTM10/1基因型显示的BPDE-DNA加合物浓度比GSTM11/1基因型略高(分别为12.3和12.4 ng/μg,而GSTM11/*1基因型为7.8 ng/μg)。银屑病患者中BPDE-DNA加合物与GSTM1基因型之间的非显著关系可能与GT中使用的相对低剂量CCT和短期UV-R暴露有关。

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