ABSTRACT
Objective:To explore the differences of regulator of G protein signaling 4 (RGS4) gene polymorphisms and methylation between schizophrenia and healthy controls, as well as the association between gene polymorphisms and methylation.Methods:A total of 129 schizophrenia patients and 131 healthy controls from Southen Fujian were enrolled in this study. The peripheral blood DNA of all the subjects was extracted.The three polymorphic loci of RGS4 (rs10759, rs12753561 and rs951436) were amplified, sequenced, and then genotyped. In addition, 32 subjects were randomly selected from the two groups respectively and the gene methylation level of RGS4 was detected by sequencing after bisulfite treatment. SPSS 20.0 software was used for data analysis. The χ2 test and independent sample t-test were used to analyze the difference of gene methylation of RGS4.Two-way ANOVA was used to analyze the association between RGS4 gene polymorphism and methylation. Results:There were three genotypes of AA, AC and CC for rs10759 locus in the subjects of patient group and control group. And the distribution difference of genotypes between the two groups was statistically significant ( χ2=6.431, P=0.040), but there was no significant difference in allele frequency( χ2=1.270, P=0.260). For rs12753561, there were three genotypes of GG, GT and TT, and their distribution of genotypes was significantly different ( χ2=6.217, P=0.045). There was no significant difference for the allele frequency for rs12753561( χ2=0.021, P=0.885). For rs951436, there were three genotypes of AA, AC and CC, and there were no significant difference in genotype distribution and allele frequency distribution between the two groups( χ2=0.008, 0.007, both P>0.05). Methylated CpG sites were found in 26 patients and 27 healthy controls, and these were no significant difference between the two groups( χ2=0.110, P=0.740). There was no significant difference ( t=-0.318, P=0.752) of individual methylation rate (number of methylation sites/10) between schizophrenia patient group (0.24±0.11) and healthy control group (0.26±0.18). There was also no significant difference of methylation rate between male and female in both groups(both P>0.05). Finally, there was no significant difference of individual methylation rate among rs10759, rs12753561 and rs951436 genotypes (all P>0.05). Conclusion:RGS4 rs10759 and rs12753561 genotypes may be involved in schizophrenia, while RGS4 gene methylation has no association with schizophrenia. In addition, RGS4 gene polymorphism has no association with the methylation in the current experimental setting.
ABSTRACT
Objective To investigate the antagonism role of adenosine and its agonists in hypoxia-induced right ventricular hypertrophy in rat.Methods Fifty-six Sprague-Dawley rats were randomly divided into 7 groups,including normoxia control group,hypoxia control group,hypoxia plus adenosine group,hypoxia plus adenosine A1 receptor agonist(CPA) group,hypoxia plus adenosine A2 receptor agonist(NECA) group,hypoxia plus CPA and adenosine A1 receptor antagonist (DPCPX) group,hypoxia plus NECA and adenosine A2b receptor antagonist(MRS1754) group.There were 8 rats in each group.Rats in all groups except normoxia group were exposed to hypoxia for 21 days(oxygen volume concentration of 95-105 mL/L).On the 7th day,agents were continuously delivered with micro osmotic pump for 14 days.At the end of the experiment,both right ventricular to left ventricular plus septum weight[RV/(LV + S)] ratio and right ventricular to body weight (RV/BW) ratio were measured,and the expression of myosin heavy chain (β-MHC) mRNA and regulator of G protein signaling 4(RGS4) mRNA in the right ventricular myocardial tissues were detected by PCR method.Results 1.After chronic hypoxia for 21 days,the ratios of RV/(LV + S) and RV/ BW in the hypoxia control group were significantly higher than those in the normoxia control group respectively(all P ＜ 0.001) ; the ratios of RV/(LV + S) and RV/BW in the hypoxia plus adenosine group and hypoxia plus CPA group were significantly lower than those in the hypoxia control group (all P ＜ 0.05) ; the ratio of RV/(LV + S) in the hypoxia plus NECA group was significantly lower than that in the hypoxia control group(P ＜ 0.05).2.The expression level of β-MHC mRNA in the hypoxia control group was higher than that in the normoxia control group(P ＜ 0.001),and the expression level of RGS4 mRNA in the hypoxia group was lower than that in the normoxia group(P ＜ 0.001) ; the expression levels of β-MHC mRNA in the hypoxia plus adenosine group,hypoxia plus CPA group,or hypoxia plus NECA group were lower than those in the hypoxia control group (all P ＜ 0.001) ; the expression levels of RGS4 mRNA in hypoxia plus adenosine group,hypoxia plus CPA group,and hypoxia plus NECA group were higher than those in the hypoxia control group(all P ＜ 0.001).Conclusions Adenosine can attenuate hypoxia-induced right ventricular hypertrophy through RGS4 mediated pathway.