| Objective: 1. To study the effect of complement protein fragments dissolved on the formation of brain edema after ICH; 2. To study the impact of Camostat Mesilate for protein fragments dissolved from complement activation; 3. To observe the inhibition effect of Camostat Mesilate on the formation of brain edema after ICH; 4. To explore the mechanism of the inhibition effect of Camostat Mesilate on the formation of brain edema after ICH. Methods: 1. To divide SD rats into groups: 90 SD rats were randomly divided into three groups (30 per group), named sham operation group, ICH control group and Camostat Mesilate treating group. Every group had six observations set: 12h, 1d, 2d, 3d, 7d, 14d after operation. There were five animals at each observation set. 2. Models and administration. Autologous blood was injected in the right caudate nucleus of rats by the solid directional instrument. The successful estimate of the model would be done at 2h post operation. Camostat Mesilate (50mg/kg·d) was injected in the abdominal cavity of the rats in Camostat Mesilate treating group , equal volume normal saline was injected in the abdominal cavity of the rats in ICH control group everyday. The surgical procedures of Sham-operated group like with the ICH control group, but autologous blood wasn't injected in the caudate nucleus of rats, indwelling needle for about 10 minutes. 3. Neurobehavioral deficit outcome would be estimated at each observation time point.4. Water contents of brain tissue circum-hematoma would be detected by dry-wet weigh method at each observation time point. 5. Use radiation immunity method to detect IL-1β,IL-6 and TNF-αlevel at each observation time point. 6. Use colorimetric method to detect nonheme iron of brain tissue circum-hematoma at each observation time point. 7. Use immunohistochemistry to detect Complement C5a and C9 fragment of brain tissue circum-hematoma at each observation time point. Results: 1. Neurobehavioral deficit outcome at various stages in sham operation group, ICH control group and Camostat Mesilate treating group: Neurobehavioral deficit outcome at 12h～14d in ICH control group were significantly lower than those at corresponding time points in sham operation group (P<0.05). Neurobehavioral deficit outcome at 2d were significantly lower than those at other time points in ICH control group (P<0.05), but not differ significantly from those at 3d (P>0.05). Neurobehavioral deficit outcome at 12h～7d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in sham operation group (P<0.05). Neurobehavioral deficit outcome at 2d～7d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in ICH control group (P<0.05). 2.Water contents of brain tissue circum-hematoma at various stages in sham operation group, ICH control group and Camostat Mesilate treating group: Water contents of brain tissue circum-hematoma at 1d～7d in ICH control group were significantly higher than those at corresponding time points in sham operation group (P<0.05). Water contents of brain tissue circum-hematoma at 2d were significantly higher than those at other time points in ICH control group (P<0.05), but not differ significantly from those at 3d (P>0.05). Water contents of brain tissue circum-hematoma at 1d～3d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in sham operation group (P<0.05). Water contents of brain tissue circum-hematoma at 1d～7d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 3. IL-1βlevel at various stages in sham operation group , ICH control group and Camostat Mesilate treating group: IL-1βlevel at 12h～3d in ICH control group were significantly higher than those at corresponding time points in sham operation group (P<0.05). IL-1βlevel at 2d were significantly higher than those at other time points in ICH control group (P<0.05). IL-1βlevel at 12h～3d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in sham operation group (P<0.05). IL-1βlevel at 2d～3d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 4. IL-6 level at various stages in sham operation group, ICH control group and Camostat Mesilate treating group: IL-6 level at 1d～7d in ICH control group were significantly higher than those at corresponding time points in sham operation group (P<0.05). IL-6 level at 2d were significantly higher than those at other time points in ICH control group (P<0.05). IL-6 level at 1d～3d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in sham operation group (P<0.05). IL-6 level at 2d～3d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 5. TNF-αlevel at various stages in sham operation group , ICH control group and Camostat Mesilate treating group: TNF-αlevel at 12h～3d in ICH control group were significantly higher than those at corresponding time points in sham operation group (P<0.05). TNF-αlevel at 2d were significantly higher than those at 7d, 14d in ICH group (P<0.05) , but not differ significantly from those at 12h, 1d, 3d (P>0.05). TNF-αlevel at 1d～2d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in sham operation group (P<0.05). TNF-αlevel at 1d～2d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 6. The content of nonheme iron at various stages in sham operation group , ICH control group and Camostat Mesilate treating group: the content of nonheme iron at 3d～14d in ICH control group were significantly higher than those at corresponding time points in sham operation group (P<0.05). The content of nonheme iron at 14d were significantly higher than those at other time points in ICH control group (P<0.05). The content of nonheme iron at 3d～14d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in sham operation group (P<0.05). The content of nonheme iron at 3d～14d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 7. The expression of complement C5a at various stages in sham operation group , ICH control group and Camostat Mesilate treating group: The number of positive cells of complement C5a at 12h～14d in ICH control group were significantly higher than those at corresponding time points in sham operation group (P<0.05). The number of positive cells of complement C5a at 2d were significantly higher than those at other time points in ICH control group (P<0.05). The number of positive cells of complement C5a at 12h~14d in Camostat Mesilate treating group were significantly higher than those at corresponding time points in sham operation group (P<0.05).The number of positive cells of complement C5a at 12h～7d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 8. The expression of complement C9 at various stages in sham operation group, ICH control group and Camostat Mesilate treating group: sham operation group at different time points were no expression of complement C9. The number of positive cells of complement C9 at 1d were significantly higher than those at other time points in ICH control group (P<0.05). The number of positive cells of complement C9 at 12h～7d in Camostat Mesilate treating group were significantly lower than those at corresponding time points in ICH control group (P<0.05). 9. Linear correlation : (1) The ICH control group: neurobehavioral deficit outcome is siginificantly negatively correlated with water contents of brain tissue circum-hematoma, IL-1βlevel , IL-6 level, TNF-αlevel, the number of positive cells of complement C5a and the number of positive cells of complement C9. Water contents of brain tissue circum-hematoma was significantly positively correlated with IL-1βlevel, IL-6 level,TNF-αlevel, the number of positive cells of complement C5a and the number of positive cells of complement C9. The number of positive cells of complement C5a was significantly positively correlated with IL-1βlevel, IL-6 level and TNF-αlevel .The number of positive cells of complement C9 was significantly positively correlated with IL-1βlevel , IL-6 level and TNF-αlevel . (2)The Camostat Mesilate treating group: neurobehavioral deficit outcome was significantly negatively correlated with water contents of brain tissue circum-hematoma, IL-1βlevel , IL-6 level, TNF-αlevel, the number of positive cells of complement C5a and the number of positive cells of complement C9. Water contents of brain tissue circum-hematoma was significantly positively correlated with IL-1βlevel, IL-6 level, TNF-αlevel, the number of positive cells of complement C5a and the number of positive cells of complement C9. The number of positive cells of complement C5a was significantly positively correlated with IL-1βlevel , IL-6 level and TNF-αlevel .The number of positive cells of complement C9 was significantly positively correlated with IL-1βlevel, IL-6 level and TNF-αlevel. Conclusion: The rat models with ICH were made by autologous blood, the way was simple and made easily, the changes of pathophysiology of rat models were almost the same as the patients with ICH, the model could be used generally. After intracerebral hemorrhage, complement dissolved fragments that generated from the brain tissue around the hematoma complement cascade increased the inflammatory response, membrane attack complex damaged red blood cells, red blood cell metabolites involved in the formation of brain edema. After intracerebral hemorrhage, the application of Camostat Mesilate special to improve neurological function, reduce the brain edema. Camostat Mesilate blocked complement cascade by inhibiting the proteasome so as to reduce the inflammatory response and reduce the destruction of red blood cells. Camostat Mesilate exerted treatment effect on the formation of brain edema after intracerebral hemorrhage.
|
PDF Full Text Request