| Background The diagnosis and treatment of substance addiction relies mainly on the experience of doctors for lack of objective biomarker.Metabolomics is a new technology for studying the metabolic network of biological systems which can reflect the comprehensive metabolic status and the pathophysiological processes of the organism.It is expected that biomarkers of substance dependence will be discovered through this technology.For the treatment of synthetic drugs,r TMS is a mild and non-invasive physical therapy.It has been found to be effective in reducing craving,but the biomarker for efficacy is absent,and the influencing factor is also unknown.For the treatment of traditional opioids,Methadone maintenance treatment(MMT)is the main treatment for opioid dependence,previous study found lots of efficacy influencing factors,however,the results were not consistent.Objective 1.Explore metabolomics character of methamphetamine dependent patients.Build a network of possible regulatory mechanisms for differential metabolites and validate the expression of candidate regulatory genes and enzymes.2.Explore the candidate biomarker of r TMS treatment of methamphetamine dependence.And explore the influencing factors of r TMS.3.Explore the influencing factors of MMT treatment of heroin dependence.Methods 1.40 methamphetamine dependence patients were enrolled,and 38 healthy controls were recruited.240 patients with heroin dependence who were treated with MMT were enrolled.General information was collected and related scales were assessed.2.Gas chromatography-mass spectrometry(GC-MS)was used to detect the serum metabolomics of methamphetamine dependence subjects.Bioinformatics approach was used to establish a regulatory network model.The expression of the regulatory enzymes and genes in this model were detected by ELISA and Q-PCR.3.Methamphetamine dependence patients were randomly divided into two groups,r TMS intervention and pseudo-stimulation were given respectively.GS-MS was used to detect the serum metabolomics characteristics before and after intervention.Characteristic therapeutic biomarker and influencing factors of r TMS were analyzed.4.The MMT outpatients who were enrolled were followed up for 6 months.The correlations between OPRM1,ABCB1 gene polymorphism and shedding/negative rate of morphine urine test were analyzed.The influencing factors of MMT efficacy were analyzed.Results 1.In methamphetamine dependence patients,compared with control subjects,35 differentially expressed metabolites were detected(p<0.05),of which 21 metabolites can be queried pathways and were distributed in four significantly altered pathways:(1)Arginine synthesis,(2)alanine,aspartic acid,glutamate metabolism,(3)cysteine and methionine metabolism,(4)purine metabolism(p <0.05).2.Selected the differential metabolits in “alanine,aspartic acid,and glutamate metabolism pathway” to establish a regulatory network model.A total of 13 regulatory genes were found,which regulated the expression of four key enzymes through 29 intermediate proteins.And ultimately affect metabolites expression.3.The expression of the regulatory enzymes in this model was detected by ELISA,and no significant difference between the two groups(p>0.05)was found.The expression of the regulatory genes in this model was detected by Q-PCR,and five genes were found to be significantly different expressed which were DLG2,PLA2G4,PDE4 D,PDE4B,and EPHB2(p<0.05).4.After intervention of r TMS,the craving level of the dependence subjects decreased,and there were 8 metabolites expression changed significantly(p<0.05),they were atocopherol(1.3 times higher than that before intervention),5-hydroxylysine(0.8 times),decanoylcarnitine(1.8 times),glyceric acid(1.2 times),malic acid(1.3 times),Caproic acid(1.4 times),fumaric acid(1.2 times),a-ketoisovalerate(1.1 times).In which three metabolites was reversed from the baseline,they are a-tocopherol,glyceric acid,fumaric acid.These three metabolites have the potential to be the therapeutic biomarkers.Multiple linear regression analysis suggested that r TMS,atocopherol,withdrawal duration,PSS-10 score were the influencing factors of craving change.5.The allele and genotype of rs562859(OPRM1 gene)was significantly different between the maintenance group and drop out group(p<0.05).A allele / AA genotype carriers were significantly more,and G allele / GG genotype carriers were significantly less in the maintenance group(p<0.05).No significant difference was found in other SNPs of ORPM1 and ABCB1 gene between the two groups(p>0.05).In the negative rate of morphine urine test,rs3192723 GG(OPRM1 gene)carriers was significantly lower than that of GA/AA carriers;and rs562859 AA carriers was significantly higher than that of AG/GG carriers(p<0.05).No significant difference was found in other SNPs of OPRM1 and ACBC1 gene between two groups(p>0.05).Logist regression analysis showed that the rs2032582(GG)(ABCB1 gene)carriers,age of first drug use,daily dose of heroin,duration of nicotine use,and unplanning score of BIS affected MMT treatment compliance.The rs3192723(GG)and rs2236259(TT)cairriers(OPRM1 gene)affected the positive rate of morphine urine test.Conclusions 1.Methamphetamine effects a long-term time on humans,L-glutamine,pyruvic acid,L-aspartate were the candidates of methamphetamine dependence biomarkers;Atocopherol,glyceric acid,fumaric acid were expected to be the candidates of r TMS treatment biomarkers.2.The glutamate metabolic pathway is regulated by a wide range of genes,including signal transduction,nervous system development,cell membrane transport,synapses development,plasticity and stability.Which indicated that methamphetamine has a wide range of effects on the human body and is not limited to several neurotransmitters.3.OPRM1,ABCB1 genotype and age of first drug use,daily dose of heroin,duration of nicotine use,impulsivity can affect the efficacy of MMT. |
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