The Morphology Detection Of Gap Junction After Cerebral Vasospasm Caused By Subarachnoid Hemorrhage In Rats

Objective：On the bases of reliable and successful animal model, the diameter of the basilarartery can be measured by india ink staining cerebrovascular casting method。Thealteration of distribution of Cx43and Cx40and their ultrastructure can be observedby the using of of laser scanning confocal microscope and transmission electronmicroscopy.The goal of these methold is to futher explore the potential role of gapjunction after Cerebral vasospasm caused by subarachnoid hemorrhage in rats.Methold:1、Choosing adult SD rats as a animal model and establishing rat,s CVS modelafter secondary SAH by secondary injections of arterial blood of rats into its,cisternamagna。2、Model checking: CVS cerebral vascular casting method is used to determinebase artery diameter of the normal group and SAH3d、7d、14d groups to confirmecerebrovascular spasm (CVS).3、The distribution of Cx40and Cx43was measured by the application ofimmunofluorescence histochemical staining technique and the laser scanningconfocal microscope in each layer of the cerebral basilar artery,and then the digitalImage was stored in the computer for Image Analysis: generally used Image Analysisprocedure to analyze images, Cx40and Cx43pixel density, simultaneousdetermination Cx40and Cx43fluorescent spot area size, measured its pixel densityand fluorescent spot area size on behalf of Cx40and levels of Cx43, carries on thequantitative Analysis.4、By the use of scanning electron microscopy,the ultrastructure of the gapjunction and cerebral basilar artery was observed before and after the rats cerebralvasospasm caused by SAH.Conclusion:1、The obvious alteration of the distribution of Cx40and Cx43in each layer in rat have taken place between the normal cerebral basilar artery and cerebralvasospasm caused by SAH.2、Further confirmed the existence of gap junction channels between vascularendothelial cells (EC-EC) and smooth muscle cells (SMC-SMC), endothelial cellsand smooth muscle cells (EC-SMC), and the gap junctions increased obviously afterCVS in the normal group. Provides strong evidence for the potential mechanism ofcerebral vasospasm.