| objectiveThe mechanism of depression is very complex.With the gradual acceptance of acupuncture in clinical practice,There are more and more studies on the mechanism of acupuncture in the treatment of depression.To observe the effect of acupuncture on the expression of GABAAR and GABABR in frontal lobe and hippocampus of rats exposed to chronic restraint stress,and to reveal the possible mechanism and possible signaling pathway of acupuncture intervention in depressive disorder.MethodsTwenty-four male SD rats were randomly divided into blank group(N),model group(M),acupuncture group(A)and fluoxetine group(F),6 rats in each group.The rats in each group were fed for one week before the experiment.In addition to the blank group,the psychological stress model was established by chronic restraint stress combined with isolation.The specific method:using barbed wire,both ends of which are fixed by a clamp.The aim was to keep the rats breathing unobstructed,and not to tioppress the rats.Restraint stress daily from 9am to 3pm,during the period of restraint,intake and water deprivation.After the end of the shackles,the rats were put into mouse cage,free food intake.28 days in a row.①blank group(N):Rats feed,6 per cage,intake and water deprivation time for the daily 9am-3pm/d.②model group(M):Rat solitary,1 per cage,restraint stress time for the daily 9am-3pm/d,intake and water deprivation for during the period of restraint,the rest of the time free feeding water and food.③acupuncture group(A):Rat solitary,1 per cage,restraint stress time for the daily 9am-3pm/d,intake and water deprivation for during the period of restraint,the rest of the time free feeding water and food.At the same time,selecting the "Baihui"(GV20),"Yintang"(GV29)and double "Sanyinjiao"(SP6)points to acupuncture intervention,lhour before restraint stress,retaining needle for 20 minutes,once a day,continue 28 days.④fluoxetine group(F):Rat solitary,1 per cage,restraint stress time for the daily 9am-3pm/d,intake and water deprivation for during the period of restraint,the rest of the time free feeding water and food.At the same time,using fluoxetine administered intervention 1hour before restraint stress.Fluoxetine was mixed with distilled water for 1.8mg/kg and was administered by 10ml/kg/d.To reflect the quality changes and dietary status of rats.Weighing rats 1 day before modeling and fourteenth,twenty-eighth days after modeling,The weight growth(The difference of body mass between the last time point and the previous time point)was calculated with the baseline of the body weight before modeling.Training all animals to adapt to sugar water before making the model.Intake and water deprivation for 23 hours before seventh,fourteenth,twenty-first after modeling.And then conduct the sugar water consumption experiment.Give two bottles of water to every rat,one of the bottle is 1%sucrose water,another one is pure water,then take and weigh the bottles.Calculate the sugar water consumption(Sugar water preference = sugar water consumption/total liquid consumption*100%)of animals and Evaluate pleasure behavior.Results1.The results of behavioral:The value of weight growth:①Compared with the group N,the weight gain of the group M was significantly lower(P<0.01).② Compared with the group M,the weight gain of the group A was significantly increased,and it was close to the growth value of the group N(P<0.01).③ Compared with the group M,the weight gain of the group F was increased(P<0.01).④ compared with fluoxetine group,there was no significant difference in body weight increment between acupuncture group and control group(P>0.05).Sugar water preference:① Compared with group N,the sugar water preference of M rats was significantly lower than that of group(P<0.01).② Compared with group M,the sugar preference of group A was significantly increased,and it was close to that of group N(P<0.01).③ Compared with group M,the preference of sugar water of group F increased(P<0.01).④compared with fluoxetine group,there was no significant difference in the index of sugar and water preference between the acupuncture group and the control group(P>0.05).2.The results of Western Bolt:Relative expression of GABAAR/GABABR in the hippocampus:①Compared with group N,the GABAAR and GABABR protein levels in the hippocampus of group M were significantly decreased(P<0.01,P<0.01).② Compared with group M,the GABAAR and GABABR protein levels in the hippocampus of group F were significantly increased(P<0.05,P<0.05).③ Compared with group M,the GABAAR protein levels in the hippocampus of group A were significantly increased,while the GABABR not(P<0.05;P>0.05).④Compared with fluoxetine group,the content of GABAAR protein in hippocampus of acupuncture group was not significantly different(P>0.05).Relative expression of GABAAR/GABABR in frontal lobe:① Compared with group N,the GABAAR and GABABR protein levels in frontal lobe of group M were significantly decreased(P<0.01,P<0.01);② Compared with group M,he GABAAR and GABABR protein levels in frontal lobe of group F were significantly increased(P<0.01,P<0.05);③Compared with group M,he GABAAR and GABABR protein levels in frontal lobe of group A were significantly increased(P<0.01,P<0.01).④ compared with fluoxetine group,the contents of GABAAR and GABABR in the frontal cortex of the acupuncture group were not significantly different(P>0.05,P>0.05).Conclusion1.Compared with the blank group,the weight growth value and the sugar water preference index of the model group decreased significantly,and the clinical manifestations were similar to those of the patients with depression.2.Compared with the blank group,the expression of GABAAR/GABABR protein in hippocampus and frontal lobe decreased in the model group,indicating that the onset of depression is related to the content of GABAAR/GABABR protein in hippocampus and frontal lobe.3.Compared with the model group,the weight increase value and the sugar water preference index of the rats in the acupuncture group are all increased,indicating that acupuncture can improve the depressive state of the gastrointestinal function and the loss of pleasure.4.Compared with the model group,the expression of GABAAR protein in hippocampus and GABAAR/GABABR in frontal lobe increased in acupuncture group,indicating that the antidepressant effect of acupuncture may be achieved by increasing the protein expression level of GABAAR in hippocampus and GABAAR/GABABR in frontal lobe.5.Compared with the model group,the expression of GABABR protein in hippocampus of rats was increased not obviously,this may be related to the distribution of GABABR is mainly in the cerebral cortex and relatively less in the hippocampus.It can’t exclude that GABAB receptor involvement in depression by effecting different neurotransmitters and their receptors or neurotrophic factors that related the occurrence of depression.6.Compared with the fluoxetine group,there was no significant difference in the expression of GABAAR/GABABR protein in the hippocampal GABAAR and the frontal cortex of the rats in the acupuncture group,indicating that the antidepressant effect of the acupuncture and fluoxetine was homosexual. |
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