谷胱甘肽S-转移酶基因多态性与特发性男性不育患者氧化应激水平的关系

Relationship between Glutathione S-transferase Gene Polymorphism and Oxidative Stress Level in Idiopathic Male Infertility Patients

目的:分析特发性男性不育患者谷胱甘肽S-转移酶(GSTs)基因GSTT 1、GSTM 1及GSTP 1基因多态性与氧化应激水平及精子DNA氧化损伤的关系。方法:收集246例特发性男性不育患者的静脉血,提取全血基因组DNA,采用聚合酶链反应(PCR)对GSTT 1及GSTM 1基因进行分型,采用限制性片段长度多态性聚合酶链反应(PCR-RFLP)对GSTP 1基因进行分型;采集患者精液,酶联免疫吸附法(ELISA)检测精浆中丙二醛(MDA)、一氧化氮(NO)水平及总抗氧化能力(T-AOC),同法检测精子DNA中8-羟基脱氧鸟苷(8-OH-dG)水平。结果:GSTM 1(-)及GSTT 1(-)组中精浆NO及精子DNA中8-OH-dG水平分别显著高于GSTM 1(+)及GSTT 1(+)组(P<0.01),GSTM 1/GSTT 1(-/-)组精浆MDA、NO及精子DNA中8-OH-dG水平高于GSTM 1/GSTT 1(+/+)组(P<0.05),而精浆T-AOC低于GSTM 1/GSTT 1(+/+)组(P<0.05),GSTP 1(A/G+G/G)组中NO及精子DNA中8-OH-dG水平高于GSTP 1(A/A)组(P<0.05),而T-AOC显著低于GSTP 1(A/A)组(P<0.01)。结论:GSTs的GSTT 1、GSTM 1及GSTP 1基因突变可增加特发性男性不育患者精浆氧化应激水平,降低精浆总抗氧化能力,导致精子DNA氧化损伤。

Objective:To analyze the relationship between glutathione S-transferase(GST)gene polymorphisms including GSTT1,GSTM1 and GSTP1 gene,oxidative stress levels and sperm DNA oxidative damage in idiopathic male infertility patients.Methods:The venous blood from 246 idiopathic male infertility patients was collected,and whole blood genomic DNA was extracted.GSTT1 and GSTM1 genes were genotyped by polymerase chain reaction(PCR).Restriction fragment length polymorphism polymerase chain(PCR-RFLP)was used to classify GSTP1 gene.The levels of malondialdehyde(MDA),nitric oxide(NO)and total antioxidant capacity(T-AOC)in seminal plasma were measured by enzyme-linked immunosorbent assay(ELISA).8-hydroxydeoxyguanosine(8-OH-dG)levels were detected in sperm DNA.Results:The levels of seminal plasma NO and 8-OH-dG were significantly higher in GSTM1(-)and GSTT1(-)groups than those in GSTM1(+)and GSTT1(+)groups,respectively(P<0.01).Seminal plasma MDA,NO and 8-OH-dG were higher in GSTM1/GSTT1(-/-)group than those in GSTM1/GSTT1(+/+)groups(P<0.05),while seminal plasma T-AOC was lower than that in GSTM1/GSTT1(+/+)groups(P<0.05).The levels of seminal plasma NO and 8-OH-dG were significantly higher in GSTP1(A/G+G/G)group than in those in GSTP1(A/A)group(P<0.05),while T-AOC was significantly lower than that in GSTP1(A/A)group(P<0.01).Conclusion:GSTT1,GSTM1 and GSTP1 gene mutations can increase the level of seminal plasma oxidative stress in idiopathic male infertility patients,reduce the total antioxidant capacity of seminal plasma,leading to oxidative damage of sperm DNA.