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Objective: widely gap junctions (gap junction GJ) stored in the cardiovascular system, the change of the gap junction channel function play an important role in many cardiovascular and cerebrovascular diseases. On the basis of the preliminary studies, the experimental immuno precipitation technique of subarachnoid hemorrhage (SAH) cerebral vascular changes in gap junction channel types, to lay the foundation for more in-depth study of cerebrovascular GJ channels participate in the mechanism of CVS after SAH may CVS to find a new way to more effective preventive. Method: 1. Establish rabbit secondary CVS after SAH model: New Zealand white rabbits were 3% sodium pentobarbital (1ml/kg) anesthesia after cisterna magna puncture injection not heparinized the fresh autologous arterial blood (0.5ml/kg,), sober After normal feeding 24 hours after the second blood injection is repeated. 2 detection of animal CVS model: divided into the sham injection group and the SAH group, in the 7d line twice before the establishment of the model and modeling DSA angiography, comparing the experimental group, the change of the diameter of the basilar artery; quadratic contrast, after the heart whole brain perfusion fixed for HE staining, light microscope morphological changes of the vascular wall. GJ protein changes: each group of animals row DSA were sacrificed after cerebral angiography separation of the basilar artery, the application of co-immunoprecipitation technology for the purification of the corresponding protein antibody precipitated Cx43, Cx45, Cx40, Cx3 Western Blot, respectively. change detection basilar artery of Cx43, Cx45, of Cx40, Cx37 and base it three protein interaction, suggesting that these connexin up the same type of channel or abnormal channel change. Results: 1. Successfully established rabbit secondary CVS after SAH model: DSA angiography before and after the normal control group modeling the basilar artery diameter ratio of 97.2% ± 1.5%, and the basilar artery diameter ratio was 63.7% ± 7.1 before and after the SAH-7d group modeling %, with a significant difference (p <0.05) compared with the control group. Narrow vascular lumen light microscope spasm and wall thickening; endothelial cells Partial necrosis induced exposed internal elastic lamina, cytoplasmic vacuolization see, part of the cell nuclei hyperchromatic pyknotic; internal elastic lamina ripple like curling, fracture; smooth muscle cell hypertrophy, degeneration and associated necrosis. SAH group Cx43 derived protein precipitation of Cx43, Cx45, Cx37 expression was higher than that in the sham injection group increased significantly, while the SAH group Cx40 expression was higher than that in the sham injection group significantly reduced; similar to Cx45 protein precipitation can be drawn SAH Group Cx45, Cx43, Cx37 expression significantly increased compared with the sham injection group significantly reduced compared with the sham injection group the SAH group Cx40 expression; Cx40 SAH group derived protein precipitation of Cx43, Cx45, Cx37 and of Cx40 expression was higher than that in the sham injection group were significantly reduced; to Cx37 to precipitate proteins derived SAH group of Cx43, Cx45, Cx37 expression significantly increased compared with the sham injection group, while the SAH group Cx40 expression compared with the sham injection group significantly reduced. 3. Cx43 and Cx43, Cx45 and Cx45, Cx37 and Cx37 formed the same type of gap junction channels increase in the normal group after SAH. Cx40 and Cx40 formed the same type of gap junction channels is less than normal after SAH group. Shaped Cx37 and Cx45, Cx37 and of Cx43, Cx45 and Cx43 gap junction channel than the normal group increased after SAH. Shaped Cx37 and of Cx40 of Cx40 with Cx43, of Cx40 and Cx45 gap junction channel is less than normal after SAH group. Conclusion: in CVS the process, the extent of interaction of a variety of GJ protein (Cx37, Cx40, Cx43, Cx45) occurred changes corresponding to the formation of a variety of the same type of slot connected to the channel or shaped gap junction channel quantity changes, indicate that GJ The change of the channel may be involved in the occurrence of CVS after SAH.
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