Experimental Study The Effect Of Application Of Monoclonal Antibody Against CD34 As Means Of Prevention And Treatment On CVS And Cerebral Protection After SAH In Rats

PartⅠ: The expression of CD34 in BA walls following SAH in ratsObjective: To investigate the expressions of CD34 in BA(basilar artery) walls following subarachnoid hemorrhage (SAH) in rats and to study the relationships with cerebral vasospasm (CVS).Methods: 51 health male Sprague-dawley(SD) rats were assigned randomly into following groups: Normal group, Control group(injected normal sodium through cisterna magna only), SAH group. The control and SAH groups then were assigned into 5 subgroups according to time points(1d,3d,5d,7d and 10d,Control group: n=3; SAH group: n=6). Once autologus arterial blood injection through cistema magna was underwent to establish SAH model in rats. The morphologic changes of BA were examined with hemotoxylin and eosin(HE) staining, and the expressions of CD34 in BA walls following SAH at different time point were detected by immunohistochemistry method.Results: The pathological changes including arterial narrowing, thickness of arterial wall, reductus of intima and hypertrophy of smooth muscle cells could be observed and such changes progressed with time and were more obvious on 3rd day and 5th day. The luminal inner diameter of BA on 3th day after SAH was about 60% of the control group's, and the wall thickness of BA was about double times of the control group's. The manifestation on 10th day was near to the control group.The expression of CD34 increased in SAH group, and reached to the peak on 3rd day and 5th day, then decreased. The level on 10th day was similar to the control group.Conclusions: 1. The pathological changes including morphological changes, arterynarrowing, thickness of arterial wall suggests that CVS happens after SAH. 2. The expression of CD34 in the BA walls has changes and been concerned with CVS. It suggests that CD34 may participate the pathological course of vasospasm.Part II: Experimental study of the effect of monoclonal antibody against CD34 on CVS after SAHObjective: To evaluate the effect and potential mechanism of monoclonal antibody against CD34 on CVS after SAH.Methods: 30 health male Sprague-dawley(SD) rats were assigned randomly into following groups: Normal group, SAH group,Vehicle group(injected normal sodium through cisterna magna only),Low dose monoclonal against CD34 injection group(LD), High dose monoclonal against CD34 injection group(HD).The"one hemorrhage"model of vasospasm was performed. We administered increasing doses of a monoclonal antibody against rat CD34( the dosage in LD group is 10μg/10μl and in HD group is 20μg/10μl) directly into the cisterna magna asmeans of prevention and treatment before the blood injection, while Vehicle animals received an equal volume of NS into the cisterna magna. We observed morphological changes and the expression of CD34 and endothelial apoptosis in BA walls by light microscopy and immunohistochenistry.Results: The pathological changes in Normal and Vehicle groups including the narrowing of arterial diameter(60% of the control group), thickness of arterial wall(about 2 times of the Normal group), reductus of intima. Compared with SAH and Vehicle groups, diameter of BA in HD group was expanded,thickness of BA walls and CVS value were decreased with a statistical difference (p <0.05),while LD group had no difference (P> 0.05). And there was different between HD and LD groups. The expression of CD34 in BA walls in SAH and Vehicle groups were strong, especially in media and tunica intima , whereas it was slight in LD group and no obvious in HD group. The expression of Caspase-3 was strong in normal and vehicle groups(P<0.01), but it was slight in LD group(P<0.05) and no obvious in HD group(P<0.01).What's more, different dose of monoclonal antibody against CD34 showed different effect (P<0.05). Conclusions: Monoclonal antibody against CD34 showed good effect for prevention and treatment on CVS after SAH. At the same time that monoclonal monoclonal antibody against CD34 may prevention and treatment on CVS by inhibited Caspases enzymes leading to reduce endothelial apoptosis.Part III The Effect of Protection of Monoclonal Antibody Against CD34 on brain injury caused by SAH in RatsObjective: To evaluate the effect of monoclonal antibody against CD34 on reduced apoptosis of the temporal lobe's cortical and the hippocampus neuron's which caused by SAH.,and the effect for protection on ischemic brain.Methods: 30 health male Sprague-dawley(SD) rats were assigned randomly into 5 groups. The"one hemorrhage"model of vasospasm was performed. We administered increasing doses of a monoclonal antibody against rat CD34( the dosage in LD group is 10μg/10μl and in HD group is 20μg/10μl) directly into the cisterna magna asmeans of prevention and treatment before the blood injection, while Vehicle animals received an equal volume of NS into the cisterna magna. We observed the expression of Caspase-3 in the temporal lobe's cortical and the hippocampus by immunohistochenistry.Results: The expression of Caspase-3 of the temporal lobe's cortical and the hippocampus in SAH and Vehicle groups were strong, and it expressed in neuronal cytoplasm which is dark(p <0.01) , whereas it was slight in LD group(p <0.05) and no obvious in HD group(p <0.01). What's more, different dose of monoclonal antibody against CD34 showed different effect (p <0.05).Conclusions: Monoclonal antibody against CD34 showed good effect for prevention and treatment on brain injury caused by SAH and was like to reduce neuronic apoptosis.