Correlative Research Of Expression Change Of PKC, NF-kB And ICAM-1as Well As The Relation With The Cause Of CVS After SAH

PartⅠThe clinical research of the variation of NF-kB,sICAM-1,sVCAM-1 content of blood in patients with ruptured aneurysmal subarachnoid hemorrhage Objective: To explore the relations between dynamic content change of NF-kB, sICAM-1, sVCAM-1 in blood of patients with SAH and the progress of the symptom. Methods: 30 cases of ruptured intracranial aneurysms were studied in hospital in 48h after the bleeding. The volume of SAH were evaluated according to Fisher CT grade and the severity of symptom were evaluated according to Hunt and Hess grade. Position of aneurysms were identified by DSA and the CVS were confirmed by TCD. Venous blood were analysed 3, 5, 7, 9, 11d after intracranial aneurysms ruptured. Anticoagulating plasm and outside monocyte were separated, stored in -80℃refrigerator and measured together. The control group were 20 healthy people. Results: The expression of NF-kB got to top in 3d, sustained to 5d, then decreased and basically back to normal in 9d. The expression of sICAM-1,sVCAM-1 got to top in 5d, began to decrease in 7d, back to normal in 11d. The mean data of NF-kB in 3, 5d and sICAM-1,sVCAM-1 in 5,7d were significantly higher than the control group. There were significant difference between CVS and non-CVS groups. These closely related to the occurrence and progress of disease. Content changes of NF-kB after SAH were significantly earlier than sICAM-1, sVCAM-1 in 3~5ds. Conclusion: Variation of NF-kB, sICAM-1, sVCAM-1 in blood of patients with SAH had correlation with the progress and degree of CVS. All means that ICAM-1 and VCAM-1 induced by NF-kB played important roles in the occurrence and progress of CVS. Part ⅡThe expression of PKC, NF-kB, ICAM-1 and VCAM-1 in the basilar artery of rabbits Objective: To discuss the existence of PKC, NF-kB, ICAM-1and VCAM-1 in the occurrence and progress of CVS in the basilar artery of rabbits and the relations of them. Methods: 110 Newzealand pure rabbit from 4~6M were afforded by the animal center of SuZhou University medical college without the limitation of gender with the mean weight of 2.5±0.4kg. Models of SAH and CVS weresuccessfully made via double blood injection into major cistern and basilar artery were separated , preserved under zero and observed collectively. Morphological methods, immuno-histochemistry, hybridization in situ as well as Western blot were employed to observe the variance of basilar artery and dynamic expression of PKC, NF-kB, ICAM-1and VCAM-1 in the wall of the BA. Results: Stenosis of BA were identified by DSA 3d after SAH and got to the top in 5d which was accordance with morphological observation and could afford foundation for the observation of patho-physiological changes of CVS. The expression of PKC, NF-kB, ICAM-1and VCAM-1 increase accompanied with the stenosis of spasmodic artery. The expression of PKC, NF-kB significantly increased 1d after double blood injection, maintained in the high level during 3~5ds and began to decrease in 7d, back to normal in 9d. The expression of ICAM-1, VCAM-1 began to increase in 1d, got to the top in 5d, and decreased in 7d, back to normal in 11d. The expression of PKC, NF-kB, ICAM-1and VCAM-1 in protein and mRNA level had difference in sequence and had close correlation with the happening and progress of CVS. Conclusion: Inflammation of spasmodic artery controlled by PKC, NF-kB and induced by ICAM-1, VCAM-1 do exist in the happening and progress of cerebral vasospasm. These relations played important functions in the happening and progress of CVS.Part ⅢThe experimental research of inhibitors of PKC and NF-kB in the treatment of CVS in basilar artery of rabbits Objective: To observe the effect of inhibitors of PKC and NF-kB in CVS by breaking the expression of adhesion molecule and other inflammatory factors in gene level, then to try to provide a new approach of treatment in CVS. Methods: The inhibitors of PKC, NF-kB were injected for 2ds through major cistern 3d after the model of CVS were made and got the BA to give a morphologic observation. Methods of immuno-histochemistry, hybridization in situ and Western blot were employed to examine the expression and effect of PKC, NF-kB, ICAM-1 and VCAM-1 in the wall of spasmodic vessels. Results: Intimas were plain under light microscope. There were no obvious ripple crimple and the endodermis cells arrayed regularly. The stenosis degree of the vessels decreased obviously and there were no evident thickness of the wall of vessels. There were obvious stenosis of vessels and obvious thickness of the walls of vessels as well as the intimas twisted inwards in control group. PKC,NF-kB,ICAM-1and VCAM-1 mainly existed and had a little expression in intimas and subintimas. The effect of PKC inhibitors were much evident. The effect of using antibody of ICAM-1 was not as good as that of PKC, NF-kB inhibitors. Conclusion: Using the inhibitors of PKC, NF-kB could restrain the expression of PKC, NF-kB, ICAM-1and VCAM-1in BA.This meant that theinhibiting treatment of PKC, NF-kB might provide some effect in preventing CVS.