The Epigenetic Mechanism Underlying Ethanol Dependence Of Wistar Rats And The Effect Of Epigenetic On The Susceptibility Of Ethanol Dependence And Epigenetic Of The Offspring

Objective(1) To study the epigenetic mechanism underlying the chronic ethanol dependence.(2) To investigate the effect of histone deacetylase on ethanol-induced CPP.Methods(1)48male and48female Wistar rats were respectively assigned to ethanol group, ethanol+sodium butyrate group, sodium butyrate group, saline group. The rats were recived CPP training and CPP test.(2) Using real-time PCR quantified htr3a mRNA.(3) ChIP assays were performed to determine the levels of specific histone modifications in htr3a gene promoter. Result(1) Compared with saline group, the CPP scores of EtOH group and EtOH+SB group were significantly higher. Compared with EtOH group, the CPP scores of EtOH+SB group were significantly higher.(2) htr3a mRNA expression levels were significantly higher in EtOH group, EtOH+SB group and SB group compared with saline group. The CPP score in EtOH group and EtOH+SB group was positively correlated with level of htr3a mRNA expression in PFC.(3) The acetylation of H3K9in htr3a promoter region in ethanol group was significantly increased, corresponding to the increase of htr3a mRNA expression compared with those in saline group. And there is no significant difference in levels of trimethylated H3K4in htr3a promoter region in PFC in each group. Sodium butyrate (a kind of HDAC inhibitor) enhanced the ethanol-induced CPP.Conclusion(1) Chronic ethanol exposure may upregulate htr3a of Wistar rats through mechanisms involving H3K9acetylation, which is associated with the mechanism underlying ethanol seeking behavior.(2) HDAC inhibitor sodium butyrate enhanced the ethalol-induced CPP. Part â…¡ The effect of parental exposure to ethanol on epigenetic and the susceptibility of ethanol dependence of offspringObjectiveTo investigate the effect of parents that exposured to ethanol on the epigenetic and the susceptibility of ethanol dependence of offspring.Methods24adult male and24female Wistar rats were randomly injected ethanol and saline for15days and experienced withdrawal for15days. Then coupled into4groups:male parent ethanol exposure+female parent ethanol exposure, male parent ethanol exposure+female parent saline exposure, male parent saline exposure+female parent ethanol exposure, male parent saline exposure+female parent saline exposure. The offspring were fed for2months, then received two different procedures:(1) Project1. Without ethanol exposure. Using real-time PCR quantified htr3a mRNA. ChIP assays were performed to determine the levels of specific histone modifications in htr3a gene promoter.(2) Project2. With ethanol exposure.offspring from each couple were received CPP training and CPP tests. Using real-time PCR quantified htr3a mRNA. ChIP assays were performed to determine the levels of specific histone modifications in htr3a gene promoter.Result(1) Project1. Compared to the htr3a mRNA expression of the offspring of Saline+Saline group, that of the offspring of EtOH+EtOH group, EtOH+Saline group, Saline+EtOH group significantly increased. The H3K9acetylation in htr3a promoter region of the offspring of EtOH+EtOH group, EtOH+Saline group, Saline+EtOH group are much higher than the offspring of Saline+Saline group. There is no significant difference in H3K4trimethylation in htr3a promoter region in each group.(2) Project2. Compared with the the CPP score of offspring of Saline+Saline group, that of the offspring of EtOH+EtOH group, EtOH+Saline group, Saline+EtOH group were significantly higher. Compared to the htr3a mRNA expression of the offspring of Saline+Saline group, that of the offspring of EtOH+EtOH group, EtOH+Saline group, Saline+EtOH group significantly increased. The H3K9acetylation in htr3a promoter region of offspring of EtOH+EtOH group, EtOH+Saline group, Saline+EtOH group are much higher than that of the offspring of Saline+Saline group. There was no significant difference in H3K4trimethylation in htr3a promoter region in each group. Conclusion(1) Chronic ethanol exposure induced H3K9acetylation in htr3a promoter region could inherit between generations.(2) Parental exposure to ethanol may increase the susceptibility of ethanol dependence through mechanisms involving H3K9acetylation in htr3a promoter region.