Study On The Mechanism Of Action Of CGMP In Drinking Behavior Of Rat

Object: By observing the amount of re-drinking after alcohol withdrawal in rats,investigate the effect of c GMP as a downstream pathway of 7,8-DHF on drinking behavior,and provide a new target for the clinical treatment of alcohol withdrawal,further.Methods:Experiment 1:Thirty-six male SD rats were randomly divided into control group(drinking water,n=6)and test group(drinking 20% alcohol solution,n=30).Rats in the test group were treated intermittent drinking for 28 days,and the liquid consumption was recorded daily and weight was measured weekly.After 28 days,the weight and the total fluid consumption of rats in the control group and the test group were compared.And randomly divided into test group accdording to the withdrawal time 0h,12 h,24h,48 h,72h,comparing the amount of re-drinking within 6h in each withdrawal group.Experiment 2:Eighteen male SD rats were randomly divided into the control group,the withdrawl time 0h and the meaningful drinking amount after alcohol withdrawl,and the expression of c GMP was detected by ELISA in the VTA.Experiment 3:Sixty six male SD rats were given stereotactic brain surgery to place cannula in the VTA area at 21 days after intermittent drinking,and continued to drink intermittently until 28 days after the surgery.According to experiment 1 the rats were randomly divided into sham group,ACSF group,8-Br-c GMP group,ODQ group,DMSO+ACSF group,KT5823 group,DMSO group,ODQ+ 8-Br-c GMP group,ODQ+ACSF group,KT5823+8-Br-c GMP group,KT5823+ACSF group,the amount of re-drinking was observed within 6h.Experiment 4:Twelve male SD rats were randomly divided into the meaningful drinking amount after alcohol withdrawl group and 7,8-DHF group,and the expression of c GMP was detected by ELISA.Experiment 5:Twelve male SD rats were randomly divided into7,8-DHF+ACSF group and 7,8-DHF+8-Br-c GMP group the amount of re-drinking was observed within 6h.Results:There was no significant difference between the test group and the control group in the weight difference at the end of the experiment and before the experiment(P>0.05).With the increase of drinking times,the alcohol consumption of rats finally reached a stable level,and the intermittent active drinking model was successfully established.There was no significant difference in total fluid consumption between the test group and the control group(P>0.05).In experiment 1,compared with alcohol withdrawal group for 0h,the amount of re-drinking within6 h in the withdrawal group for 72 h was statistically significant(P < 0.01).In Experiment2,compared with the control group and the withdrawal group for 0h,the content of c GMP increased more significantly in the withdrawal group for 72h(P<0.001).In experiment 3,the amount of re-drinking in 8-Br-c GMP group was significantly increased compared with sham operation group.The amount of re-drinking in ODQ group and KT5823 group was significantly decreased;ODQ+8-Br-c GMP group and ODQ+ACSF group were compared to increase the amount of re-drinking.The amount of re-drinking increased between KT5823+8-Br-c GMP group and KT5823+ACSF group,and the difference was statistically significant(P<0.05).In Experiment 4,compared with the withdrawal group for 72 h,the c GMP expression in VTA area of the 7,8-DHF group was significantly decreased,and the difference was statistically significant(P<0.01).In Experiment 5,compared with the 7,8-DHF+ACSF group,the amount of re-drinking in the 7,8-DHF+8-Br-c GMP group was increased,and the difference was statistically significant(P<0.01).Conclusions:SD rats drank 20%(v/v)alcohol solution intermittently for 28 days,and the alcohol intake finally reached a sustained and stable level,and the model of IA2 BC was successfully established.In IA2 BC model,the amount of alcohol consumed was different with abstinence time.The content of c GMP in the VTA was increased after alcohol withdrawal in rats.Exogenous injection of c GMP analogs or antagonists could increase or decrease the amount of re-drinking in rats,and c GMP could be involved in regulating drinking behavior.c GMP-PKG may act as a downstream pathway of 7,8-DHF and play an important role in the regulation of drinking behavior.