Connexins Are Involved In Studies Of Migraine Hyperalgesia

Objective: To observe the changes of connexin 36 and connexin 40 in the rat model of migraine with allodynia,and to identify the mechanism of connexin36 and connexin 40 involved the process of allodynia.Methods: 108 male SD rats were randomly divided into 9 groups(each group n=12,6 for Western Blot,the other 6 for immunofluorescence staining): blank group,sham surgery group,saline group,inflammatory soup 3 d group(IS-3D group),inflammatory soup 6 d group(IS-6D group),inflammatory soup 12 d group(IS-12Dgroup),gap junction inhibitor prevention group,gap junction inhibitor treatment group and gap junction inhibitor control group.The rat model of migraine with inflammation of the dura was induced by inflammatory soup,and the inflammatory soup was repeatedly given to induce allodynia in rats.The pain threshold of periorbital skin was detected by the Von Frey filaments.Gap junctions inhibitors carbenoxolone was treated and the trigeminocervical complex was took as specimens.Immunofluoreseence technique was used to observe the expression of connexin 36 and connexin 40.Western Blot was used to measure the protein expression levels of connexin 36 and connexin 40.Results: 1.The expression of connexin 36 and connexin 40 was found in the trigeminocervical complex.2.Connexin 36 and connexin 40 were expressed in neurons and glial cells.With the allodynia,the expression of the two proteins in the neurons and glial cells was different in IS-6D group.3.In the rat trigeminocervical complex of the IS-3D group,the IS-6D group,and the IS-12 D group,compared with the blank group,sham surgery group and saline group,the fluorescent density connexin 36 and connexin 40 increased,with statistical difference(P<0.01),the protein expression levels of connexin36 and connexin 40 increased significantly,with statistical difference(P<0.01).After giving the gap junction inhibitor,fluorescent density and the protein expression level of connexin 36 and connexin 40 were statistically significant reduced(P<0.01).4.After the gap junction inhibitor was treated,the orbital pain threshold was significantly different in the third day of inhibitor treatment group compared to the control group,with statistical significance(P<0.001);in the second day after the stimulation of inflammatory soup,there was a significant difference in the periorbital pain threshold,which was statistically significant(P<0.001)between gap junction inhibitor prevention group and control group,and the periorbital pain threshold of rats with prevention treatment did not decrease to the level of allodynia.Conclusion:1.The expression of connexin36 and connexin 40 in the trigeminocervical complex were confirmed.2.Connexin36 and connexin 40 were expressed in astrocytes or neurons of the trigeminocervical complex.3.With the appearance of allodynia in migraine rats,the proteins of connexin36 and connexin 40 were involved in the development of migraine allodynia.4.The prevention and treatment of gap junction inhibitors can significantly reduce the development of migraine allodynia.After the migraine allodynia,the treatment of gap junction inhibitors still has a relief effect.