Experimental Study Of Cx43RNAi For The Prevention Of DIND Disease In Vivo

Objective：Spontaneous subarachnoid hemorrhage (SAH) accounted for about5%of the stroke, lower than the incidence of intracranial hemorrhage and cerebralinfarction, but the case fatality rate is higher than the latter two[1]. And lack of specifictreatment at present in Global. SAH has always been a neurosurgery scholars researchhotspot and focus. Lead to DIND of pathological changes: delayed cerebral vaspasm(DIC),The main cause of DCI is delayed cerebral vasospasm and microcirculationdysfunction. Early research at our laboratory shows that gap junction may be play avery important role in delayed ischemic neurological deficit after subarachnoidhemorrhage (DIND). Application specific siRNA interference can effectively silentgap junction protein expression level, reducing the degree of cerebral vasospasm.On the basis of the previous research， this experiment intends to verify that whetherefficacious downregulation of protein Cx43would have any impacts on the occurrence of DIND to SAH or not.Methods：1、The induction of subarachnoid hemorrhage using a double blood injection, thedouble-hemorrhage model, was caused by repeating a second blood injection24hafter the first induction of subarachnoid hemorrhage using the same method.2、Identification of SAH model: divided into SAH group and normal group, theseventh day of the application of ink staining to measure the diameter of the basilarartery and Neurological score evaluate neurological deficit, identification SAHmodel.3、Detection of viral silencing efficiency：Virus treatment group，compared withSAH group and normal group,Western bloting detection virus efficiency of silence atday1,3,7,14.4、Neurological score：All neurological scores（modfication Garcia and beambalance test） were evaluated7d and14d after SAH.5、MRI: Normal group and the simple SAH module, pretreatment group,treatment group MRI detection at day7and14, including cerebral blood flow (CBF) and apparent diffusion coefficient (ADC). Respectively cerebral blood flowparameters level MRI (DWI, PWI), detection of cerebral blood flow (CBF) andapparent diffusion coefficient (ADC).Results:1、India ink staining method measuring cerebral basilar artery diameter, SAHbuilding basal artery diameter is much smaller than normal group.2、Ｖirus silence efficiency testing experiments show that the cisterna magnaadenovirus injection can effectively silence cerebrovascular Cx43expression.3、Neurological scores: modfication Garcia： SAH group compared normal groupfailed to Neurological deficit at day7，no difference at day14; All pretreatmentgroup,and All treatment group compared with SAH group, failure to improve theneurological scores.beam balance test:no difference between all group at day7and14.4、MRI results: SAH group compared with normal group of CBF and ADC valuesdecline, Virus pretreatment group and treatment group compared with SAH group,improve CBF at day7, failed to improve CBF at day14，failed to improve ADCvalues at day7and14；Carbenoxolone pretreatment, carbenoxolone treatmentgroup, the virus solvent pretreatment group, virus solvent treatment group, thecarbenoxolone solvent pretreatment group and carbenoxolone solvent treatment groupfailed to improve CBF and ADC values at day7and14.Conclusion:1、Double-hemorrhage model can reproduce the process of SAH.2、Cisterna magna injection of adenovirus, the specific downregulation of Cx43can improve CBF SAH after，suggest that can improve the outcome of SAH.