Experimental Research Of Progesterone On The Prevention And Treatment Of Delayed Cerebral Vasospasm After Subarachnoid Hemorrhage

objective:To observe the morphological change of basilar artery and the expression level change of protein kinase c(PKC) and nuclear-transcription factor-Kappa B(NF-κB) in the vascular wall of the double-hemorrhage model in Sprague-Dawley(SD) male rats, to discuss progesterone function on the prevention and treatment of delayed cerebral vasospasm(DCVS) following subarachnoid hemorrhage(SAH) and analyse its mechanism.Methods:Seventy healthy adult SD male rates were randomly divided into four groups:normal control group(NC group,n=10),sham operation group(sham group,n=20),subarachnoid hemorrhage group(SAH group,n=20),progesterone treatment after SAH group(SAHP group,n=20). Rats of NC group were done nothing but killed on day seven;rats of SAH group and SAHP group which were suffering from subarachnoid hemorrhage by the method of double-hemorrhage were killed on day three and seven respectively(each 10),there was one difference between them was that progesterone per kilo 8mg was injected by intraperitoneal injection everyday to the rats in SAHP group;rats of sham group which were used normal saline instead autologous femoral arteries blood to establish the SAH model were still killed on day three and seven(each 10);the basilar artery in the rats was removed immediately at corresponding time points.The morphological change of basilar artery was observed by HE staining and the expression level change of protein kinase c(PKC) and nuclear-transcription factor-Kappa B(NF-κB) in the vascular wall was measured by immunohistochemical staining.the datum were recorded and statistically analyzed.Results:1.hematoxylin-eosin staining(HE staining) :1) NC group and sham group:The basilar artery intima and adventitia was smooth,the structure of the endothelial cells were integrity and lined up tightly,there was no obvious ruga or breakage on the internal elastic membrane,the smooth muscle cells were in alignment,there was no obvious stricture in the vascular lumen,the vascular wall was not incrassate,and no inflammatory cell infiltration was found on the adventitia.2)SAH group:On the third day,the endothelial cells turned swell and degeneration.the internal elastic membrane began to arise ruga, breakage and uneven thickness,and smooth muscle cells were hyperplasia and confusion.there were connective tissue hyperplasia and inflammatory cell infiltration on the adventitia,the vascular lumen turned stricture and the vascular wall was incrassate.On the seventh day,all those conditions turned more and more serious.3)SAHP group:The endothelial cells turned swell and degeneration,the internal elastic membrane had ruga and crack,the smooth muscle cells and the connective tissue were hyperplasia,the adventitia had inflammatory cell infiltration on the third and seventh day,but lighter than SAH group.2. The inner diameter and tube wall thickness of basilar artery1) NC group and sham group:To compare the two groups in the seventh day,There were no difference between them(P>0.05).2)sham group and SAH group:To compare the two groups in the third and seventh day,the inner diameter of basilar artery in SAH group were less than sham group,the tube wall thickness of basilar artery were greater than sham group,there were obvious difference between them(P<0.05).3)SAH group and SAHP group:To compare the two groups in the third and seventh day,the inner diameter of basilar artery in SAHP group were greater than SAH group,less the inner diameter of basilar artery,there were obvious difference between them(P<0.05).3. Immunohistochemistry1)The expression of PKC:There were weak positive expression in the intima and smooth muscle cells in NC group and sham group,there were no difference between them on the seventh day(P>0.05).On the third and seventh day in the SAH and SAHP group,there were positive expression of PKC in the intima and smooth muscle cells,especially on the seventh day.To compare the SAH and sham group on the third and seventh day,the positive expression were both significantly enhanced in SAH group(P<0.05).To compare the SAH and SAHP group on the third and seventh day,the positive expression were both significantly subdued in SAHP group(P<0.05).2)The expression of NF-κB:There were weak positive expression in the intima and smooth muscle cells in NC group and sham group,there were no difference between them on the seventh day(P>0.05).On the third and seventh day in the SAH and SAHP group,there were positive expression of NF-κB in the intima and smooth muscle cells,especially on the seventh day.To compare the SAH and sham group on the third and seventh day,the positive expression were both significantly enhanced in SAH group(P<0.05).To compare the SAH and SAHP group on the third and seventh day,the positive expression were both significantly subdued in SAHP group(P<0.05).Conclusion:1. The enhanced expression of PKC and NF-κB in the vascular wall after subarachnoid hemorrhage may be the one mechanism of delayed cerebral vasospasm.2.Progesterone possibly restrain the activity of PKC to effect the expression of NF-κB.3. Progesterone may be restrain the activity of PKC and reduce the expression of NF-κB to relieve delayed cerebral vasospasm following subarachnoid hemorrhage.