| Objective: This study aimed to investigate Fatty acid amide hydrolase(FAAH)messenger RNA(mRNA)levels in peripheral blood mononuclear cells(PBMCs)and protein levels in plasma in Methamphetamine(METH)dependent subjects,and to analyze the FAAH 385C/A polymorphism(rs324420)in METH dependent subjects and controls.Besides,we were also evaluated its relevance to the genotypes of FAAH 385C/A polymorphism and its mRNA and protein.It was evaluated that FAAH 385C/A polymorphism was important role in METH dependence from transcription,translation and genetic polymorphism,and provided the theory foundation for the diagnosis,therapy and precaution of METH dependence.Methods: The investigation was conducted by case-control study,including 430 METH dependent subjects(cases)and 631 healthy subjects(controls),who were recruited from the Compulsory Detoxification Center of Nanchong and Affiliated Hospital of North-Sichuan Medical College from May 2013 to March 2016,respectively.All participants were of Han ethnic origin and were unrelated Chinese people by self-description.The FAAH mRNA expressions in peripheral blood mononuclear cells(PBMCs)were detected by quantitative real-time polymerase chain reaction(qPCR).The enzyme linked immunosorbent assay(ELISA)was conducted to reveal the FAAH protein levels in plasma.In addition,restriction fragment length polymorphism polymerase chain reaction(RFLP-PCR)and direct sequencinggenotyping were performed to detect the presence of the FAAH 385C/A genetic polymorphism.Statistical analyses were performed using the SPSS for Windows software package version 13.0,and a two-sided P value <0.05 was taken as the level for statistical significance.Results: In the study,84 cases and 84 controls were randomly selected for analyzing the mRNA and protein levels of FAAH,respectively.The FAAH mRNA expression levels in cases(0.5611±0.2743)were significantly decreased compared with those in controls(0.9681±0.2998)in PBMCs(P<0.001).The FAAH plasma protein levels were(4.1116±1.2659)and(2.4718±0.4939)in METH dependent subjects and controls,respectively.The FAAH plasma protein levels in METH dependent subjects were significantly higher than that those of controls(P<0.001).The FAAH 385C/A polymorphism was successfully genotyped in 430 cases and 631 controls.The genotype distribution of the FAAH 385C/A polymorphism in controls was in agreement with that expected under Hardy–Weinberg equilibrium(HWE)(P=0.10).It was indicated that our samples can represent the group in the area.The A allele frequencies of385C/A polymorphism in the METH dependent subjects and the control group were 25.58% and 17.27%,respectively.The A allele increased the METH dependent risk(P<0.001,OR=1.646,95%CI=1.332-2.034).Homogeneous genetic model,heterogeneous genetic model,dominant genetic model and recessive genetic model were used to assess the strength of association between genotypes of 385C/A polymorphism and METH dependent risk.We found that the AA,AC and AC/AA genotypes of 385C/A were observed significantly more frequently in METH dependent subjects than in controls(AA: P=0.017,OR=2.454,95%CI=1.171-2.143;AC :P<0.001,OR=1.818: 95%CI=1.404-2.353;AC/AA: P<0.001,OR=1.858,95%CI=1.444-2.319).The similar results were obtained after adjusting for age and sex.The stratified analysis accorded with demographic characteristics,body mass index(BMI)and blood routine by homogeneous genetic model(AA vs CC)and heterogeneous genetic model(AC vs CC)in cases,respectively.We found that the AC genotype were significantly different compared with CC genotype in subjects of male and higher BMI,respectively(gender: P=0.041,OR=1.901,95%CI=1.026-3.522;BMI : P=0.037,OR=1.575,95%CI=1.029-2.411).No significantly difference was found under homogeneous genetic model(P>0.05).To evaluate the functional relevance of the FAAH 385C/A polymorphisms,we analyzed the correlation between the mRNA and protein levels of FAAH and the 385 C/A polymorphism genotype in 84 healthy individuals.We failed to find that each genotype of 385 C/A affect the mRNA levels in PBMCs or plasma protein levels,respectively(P>0.05).The similar results were observed in METH dependent subjects.Conclusions: These data indicate that the mRNA and protein of FAAH may play an important role in the pathophysiological process of METH dependent,and the 385C/A polymorphism may be a genetic risk factor in METH dependent in a Chinese Han population.However,the FAAH 385C/A polymorphism did not affected its transcription and translation. |
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