Screening Of Specific Target Recognition Proteins In The Cerebral Vasospasm

Objective:Our early studies suggest that connexin43abnormally high expression of themodel of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) plays avery important role in the process, application specific siRNA interference caneffectively silence their protein content, and reduce the level of CVS. This experimentbased on this research, find cerebral vascular in CVS after SAH specific highexpression of proteins as drug identification of molecular targets to achieve targeteddrug in silent cerebrovascular Cx43aimed for the future by the foundation for thetransformation of animal experiments to clinical application.Methods:1. Twice CVS after SAH model building induced by the blood from SD ratsautologous tails artery: using the same SD rats, pump artery blood approximately0.3ml after anesthesia, Rats cerebral cisterna puncture, inject blood after clearcerebrospinal fluid outflow，following day in previous steps, preparation of tissueafter5Days for testing.2. CVS model identification: divided into model set and normal group, withrandomly selected3SD rats in the model set with the normal group. Ink stain methodfor model identification, measurement of basal artery diameter.3. CVS model and normal groups’ OPN, TNC and E-selectin expression anddistribution in brain blood vessels, brain tissue, thoracic aorta and abdominal aorta:application of immunohistochemistry and Western Blotting technical analysis CVS inmodel and normal groups’ OPN, TNC and E-selectin of protein expression in thetissue mentioned before and distribution.4. CVS model groups’ OPN and E-selectin of protein expression in the spinalcord, heart, liver, spleen, lung, pancreatic, renal, mesenteric artery, as well as retinal:application of immunohistochemistry and Western Blotting technical analysis CVSmodel groups’ OPN and E-selectin of protein expression in the tissue mentionedbefore. Results:1. Building CVS models after SAH induced by autologous arterial blood SDRats.2. Ink stain method after staining measurement of basilar artery diameter,modules is significantly less than the normal group.3. Immunohistochemical and western nlotting results shown OPN, TNC andE-selectin in brain blood vessels and brain tissue, expression in thoracic aorta andabdominal aorta and are distributed as follows: in the normal group OPN, TNC andE-selectin are low expression levels at nasilar artery. CVS model groups:(1) OPNexpression is higher in nasilar artery, and mainly in the internal coatings, externalcoatings and coated in smooth muscle cells to extracellular matrix.(2) TNC andE-selectin expression in the cerebral basilar artery elevated levels, but lower thanOPN expression levels, and mainly in the inner membrane and the outer membranelayer.(3) The three proteins are expressed in brain tissue, located within thecytoplasm.(4) OPN and E-selectin low expression in thoracic aorta and abdominalaorta.(5) TNC is higher in thoracic aorta and abdominal aorta expression, and mainlyin the intima layer and extracellular matrix coated in smooth muscle.4. Immunohistochemical and Western Blotting results shown CVS model groupOPN E-selectin in the spinal cord, heart, liver, spleen, lung, pancreatic, renal,mesenteric artery, pulmonary artery, as well as retinal expression is as follows: OPNMainly in the kidney, pancreas, mesenteric arterial intimal layer, spinal cord andretina has a higher level of expression. E-selectin of positive regional and OPN issimilar, there were few expressions of the spleen, but expression level below OPNexpression levels.Conclusion:1. TNC expression in thoracic aorta and abdominal aorta, and in CVS modelsless elevated expression in cerebral vascular than OPN and E-selectin, therefore notas a candidate target protein;2. E-selectin in the normal group and CVS model of basilar artery elevatedexpression levels than for the group OPN a lower expression levels are relatively low in vital organs of the body, may be considered as a candidate target, target recognitionOPN a slightly weaker, side effects are relatively small;3. OPN in the normal and CVS model group expression of basilar arteryincreases measurably, have positive expression in tissues and organs, can beconsidered as one of the candidates targeted, targeted recognition of its high, but theside effects are relatively large.