| Objective:1.To observe the expression of adenosine 2A receptor in different brain regions of migrainous rats.2.To investigate the effects of sleep induction on the behavior of migrainous rats and the expression of adenosine 2A receptors in different brain regions.Methods:1.Thirty adult male Sprague-Dawley rats were randomly divided into three groups,blank group A,model group B and sham group C.The rats in blank group A didn't accept any processing.The rats in group B were established a model of migraine with electrical stimulation of dura mater.The rats in sham group C were only performed electrode placement surgery without electrical stimulation.2.After electrical stimulation,the rats in each group were placed in a quiet environment.After 10 minutes,rats exhibited excessive grooming,spining and climbing cage,and the change of behavior was recorded at the same time.Immunohistochemistry and Western Blot were performed to detect the expression of adenosine 2A receptor in periaqueductal gray(PAG),trigeminal nucleus caudalis(TNC)and the trigeminal ganglion(TG).3.Thirty migraine model rats were randomly divided into three groups,control group D,sleep induction group E and sham sleep induction group F.The rats in group E were induced sleep for 1.5 hours with intraperitoneal injection of 25mg/kg diazepam.The rats in group F were given the same dose of diazepam intraperitoneally and kept awake for 1.5 hours.Group D was given the same dose of saline.4.After induction of sleep for 1.5 hours,the rats in each group were placed in a quiet environment.After 10 minutes,excessive grooming,spining and climbing cage were detected and the behavioral scores were recorded.Immunohistochemistry and Western Blot were performed to detect the expression of adenosine 2A receptor in periaqueductal gray(PAG),trigeminal nucleus caudalis(TNC)and the trigeminal ganglion(TG).Results:1.The scores of excessive grooming,spining and climbing cage in group B were significantly higher than group C(p<0.001).There was no significant difference between group A and group C.2.Immunohistochemistry results showed that the expression of adenosine 2A receptor in PAG in group B was significantly increased than that in group C(p<0.001).The expression of adenosine 2A receptor in TNC was also significantly increased(p=0.002).There was no significant difference between group A and group C.Adenosine 2A receptor expression was not detected in TG.The expression of adenosine 2A receptor in PAG,TNC and TG in group B was significantly increased than that in group C(PAG:p=0.036,TNC:p=0.042,TG:p=0.034,respectively).There was no significant difference between group A and group C.3.Compared with group F,the scores of excessive grooming and climbing cage were significantly reduced in group E(p=0.006)after sleep induction in migraine rats.There was no significant difference between group D and group F.4.Expression of adenosine 2A receptor in PAQ TNC and TG after sleep induction:immunohistochemical staining showed that the expression of adenosine 2A receptor in PAG and TNC in group E was significantly reduced than that in group F(PAG:p=0.020,TNC:p=0.038),and there was no significant difference between group D and group F.The expression of adenosine 2A receptor was not detected in TG.The results of Western Blot showed that the expression of adenosine 2A receptor in PAG,TNC and TG in group E was reduced significantly than that in group F(PAG:p-0,032,TNC:p=0.028,TG:p=0.048).There was no difference between group D and group F.Conclusions:1.Diazepam sleep induction can reduce the behavioral score of migraine rats and alleviate migraine-like symptoms.2.The increased expression of adenosine 2A receptor in migraine related brain regions suggests that the up-regulation of adenosine 2A receptor expression may be related to migraine attack in rats.3.Sleep induction in migraine rats can reduce the expression of adenosine 2 A receptor in migraine related brain regions and alleviate migraine related behavior,which suggest that the remission of migraine after sleep induction may be mediated by reducing the expression of adenosine 2A receptor. |
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