| Objective:The purpose of this study was to investigate the pathophysiological effects of the mRNA and protein of the quinone oxidoreductase 1(NQO1)in methamphetamine(METH)dependent population,and the susceptibility to NQO1 polymorphism 609C/T(rs1800566)between METH dependent population and controls.Besides,we were also analyze the relation between genetic factors and environmental factors in METH dependence,and whether the genotypes of NQO1 609C/T polymorphism influenced the transcription and translation of NQO1,in order to provide molecular biological evidence for the prevention,treatment and study of METH dependence.Methods:Demographic characteristics,laboratory data and peripheral blood samples were obtained from 392 METH dependent subjects(experimental group)and 669 healthy controls(control group).All participants were Han ethnic origin and non-consanguineous Chinese people by self-description.The quantitative real-time polymerase chain reaction(qPCR)and Enzyme linked immunosorbent assay(ELISA)were used to detect the relative expressions of NQO1 mRNA in PBMCs and protein levels in plasma,respectively.And,restriction fragment length polymorphism(RFLP-PCR)and direct sequencing genotyping were used to detect the alleles and genotypes of NQO1 609C/T polymorphism.All experimental data were analyzed by SPSS13.0,and P<0.05 was taken as statistical differences.Results:There were no significant difference in age,sex,erythrocyte count and systolic blood pressure between the experimental group and the control group(p>0.05),respectively.The body mass index(BMI)of the experimental group(20.44±5.14 kg/m~2)was significantly lower than that of the control group(22.19±3.13kg/m~2).The white blood cell count,hemoglobin and diastolic blood pressure in the experimental group were significantly higher than those in the control group(p<0.01).In our study,use qPCR and ELISA experiments to detect 84 cases and84 controls who were randomly selected,respectively.The relative expression level of NQO1 mRNA in the experimental group(3.2650±2.2943)was significantly higher than those in the controls(1.0125±0.7959)(p<0.001).The expression level of NQO1 plasma protein in experimental group(0.2368±0.1486)was significantly lower than that in control group(0.5844±0.1742)(p<0.001).The NQO1 609C/T alleles and genotypes were classified in 392experimental subjects and 669 controls.The distribution of NQO1 609C/T polymorphism genotypes in the control group was consistent with the Hardy–Weinberg equilibrium(HWE))(p=0.06),which indicated that the population in the control group was well represented.The T allele frequencies of the experimental group(46.56%)was significantly higher than those in the control group(41.78%)(P=0.032,OR=1.214,95%CI=1.017-1.450).It was indicated that the T allele increased the METH dependent risk.The frequencies of CC,CT and TT genotype of the experimental group were 24.49%,57.91%and 17.60%,respectively.The frequencies of CC,CT and TT of genotype the control group were 32.14%,52.17%and 15.69%,respectively.We found that there were statistically significant differences between experimental group and control group under heterogeneity genetic model and dominant genetic model,respectively(TC vs CC:P=0.012,OR=1.457,95%CI=1.087-1.952;TC/TT vs CC:P=0.008,OR=1.460,95%CI=1.102-1.935).There were no statistical difference between experimental group and control group under homogenous genetic model and recessive genetic model(TT vs CC:P=0.051,OR=1.472,95%CI=0.999-2.168;TT vs TC/CC:P=0.418,OR=1.147,95%CI=0.822-1.601).Multivariate logistic regression analysis was used to adjust the age and sex,while the similar results were obtained.We performed the stratified analysis accorded with the age,sex,BMI,blood pressure and blood routine in the experimental group based on homogeneity genetic(TT vs CC)and heterogeneous genetic models(TC vs CC).There was no significantly difference under homogeneous genetic model and the heterogeneous genetic model,respectively(P>0.05).The function of NQO1 609C/T polymorphism was evaluated by analying of the relationship between the polymorphism of NQO1 609C/T genotypes and the expression of NQO1 mRNA and protein in the control group,the results showed that no significant difference was found between the CT,TT and CT/TT genotypes of NQO1 609C/T polymorphism and the expression of NQO1 mRNA and protein,respectively(P>0.05).The results of the experimental group were the same as the control group.Conclusions:(1)METH dependentence may cause changes in body quality and physiological and biochemical indexes.(2)NQO1 mRNA and protein play an important role in the pathophysiological mechanism of METH dependence.(3)NQO1 609C/T polymorphism may be associated with genetic susceptibility to METH dependent population in Chinese han population,and not affected by age,sex,BMI,blood pressure and inflammatory state.(4)NQO1 609C/T polymorphism may not cause changes in NQO1transcription and translation levels. |
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