| Objective:1. To study the secondary damage in nervefunction after ICH and its mechanism.2. To explore Protective effect Ofulinastatin (UTI) on the ICH secondary brain damage and its protectivemechanism by Observe the changes of Inflammatory response, oxygenfree radicals,pathology, brain edema and apoptosis.3. To furnishtheoretical and experimental basis for the effect of UTI treatment onICH. Methods:1. Grouping: SD rats(n=90) were randomly divided intothree groups, named sham operation group(n=30), ICH group(n=30) andUTI treating group(n=30). Then each group was divided into six timepoint:12h,1d,2d,3d,7d,14d. There were five animals at each time point.2. Models.(1) The ICH group and the UTI treating group: Autologousblood was injected in the right caudate nucleus of rats by theHorseley-Clarke technique,The sham operation group had the sameprocess of operation as the other two groups, but autologous blood wasn’tinjected, needle was kept on for10minutes.3. Administration. The ICHgroup and the sham operation group:1ml normal saline was injected inthe abdominal cavity of rats everyday, the UTI treating group: UTI(50000u/kg.d) was injected in the same area, until the corresponding timepoints.4. Observation. Various observational indicators were measuredat each observation time point, Neurobehavioral deficit outcome would be estimated by using the method of Garcia,Water contents of brainwould be detected by dry-wet weigh method, IL-1β、IL-6and TNF-αlevel would be detected by radiation immunity method Use radiationimmunity method, The SOD activity were measured by xanthineoxidation(XTO) method,the MDA content were measured by xanthinethiobarbituric acid(TBA) methods, pathological changes of brain tissuecircum-hematoma would be detected by HE staining pathology method,Neurocyte apoptosis was observed around hematoma by TUNEL staining,then measured positive cells under microscope.5. Results were analyzedstatistically by SPSS17.0. Results:1. Compared with sham operationgroup, Neurobehavioral deficit outcome at12h～14d decreasedsignificantly, Water contents of brain at1d～7d were obviously higher,SOD activity at12h～7d decreased significantly, MDA level at1d～7dwere obviously higher,Water contents of brain, IL-1β level at12h～3dwas obviously higher, IL-6level at1d～7d were obviously higher, TNF-αlevel at12h～3d was obviously higher in ICH group; Compared withsham operation group, Neurobehavioral deficit outcome at12h～7ddecreased significantly, Water contents of brain at1d～3d were obviouslyhigher,SOD activity at1d～3d decreased significantly, MDA level at1d～7d was obviously higher, IL-1β level at12h～3d was obviouslyhigher, IL-6level at1d～3d were obviously higher, TNF-α level at1d～2d was obviously higher in UTI treating group (P<0.05);Compared with the ICH group, Neurobehavioral deficit outcome at2d～7d wasobviously higher, Water contents of brain at1d～7d were obviouslyhigher,,SOD activity at1d～3d was obviously higher, MDA level at1d～7d decreased significantly,IL-1β level at2d～3d decreased significantly,IL-6level at2d～3d decreased significantly, TNF-α level at1d～2ddecreased significantly in UTI treating group (P<0.05);2. in ICH controlgroup, mild brain edema was found around the hematoma at12h,Accompanied by a small amount of inflammatory cell infiltration;glialcells with mild swelling was found at1d,and the number of neuronsdecreased, disorganized,besides, Inflammatory cell infiltration could befound; a large number of inflammatory cell infiltration could be found at2d, Neutrophil granulocyte was the most, It could be seen that a highdegree of swelling and degeneration of nerve cells, showing vacuolarchanges, disappearance of Nissl bodies, nerve nucleus condensation andstain; A high degree of brain edema around the hematoma could be foundat3d, there were still a large number of inflammatory cell infiltration,degeneration and necrosis of some neurons, nuclear structure fuzzy, glialcell hyperplasia, hematoma edge scattered microglia which engulfedphagocytizing hemosiderin. Hematoma reduced significantly and glialcells hyperplasiaed apparently at7d. glial cells and vascular hyperplasiawere the most improtent at14d. The degree of the brain edema and theinflammatory cell infiltration,the proliferation of glial cells in the late in UTI treating group were lower than that in ICH group.3.Only fewTUNEL-positive cells could be found in Sham operation group;Compared with the Sham operation group, the number ofTUNEL-positive cell was obviously higher in ICH group and UTItreating group showed at each time point(P＜0.05), The ICH groupincreased more markedly, there is significant difference in1d~14d andUTI treatment group (P <0.05);A large number of TUNEL-positive cellscould be found at12h in ICH group and UTI treatment group,and that itIncreased continuously until reached the peak at3d.4.Linear correlation:At each time point in ICH group and UTI treatment group,neurobehavioral deficit outcome is negatively correlated with watercontents of brain tissue circum-hematoma, IL-1β level, IL-6level,TNF-α level, MDA level, apoptosis,but positively correlated with SODlevel(P <0.05). Water contents of brain tissue circum-hematoma waspositively correlated with IL-1β level, IL-6level，TNF-α level, MDAlevel, apoptosis. but significantly negatively correlated with SOD level(P<0.05). Conclusion:1. The rat models with ICH were made byautologous blood, the operation was simple with higher success rate, thechanges of pathophysiology of rat models were similar with the patientswith ICH.2.There were obvious brain edema、 neurological andpathological damage after ICH.3. These factors, which includedinfiltration of inflammatory cell, expression of inflammatory cytokines, generation Free radical and apoptosis, would increase Brain edema andneurological function after ICH.4. UTI can reduce brain edema andpathological damage, improve neurological function obviously after ICH.5. UTI could protect nerve cells by inhibiting the infiltration ofinflammatory cell, the expression of inflammatory cytokines, thegeneration Free radical and apoptosis. |
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