| Objective: Subarachnoid hemorrhage (SAH) is a syndrome appears afterthe blood into subarachnoid space of the brain or spinal canal mainly due torupture of intracranial aneurysm, intracranial vascular malformation and braintrauma, which is one of the high mortality and morbidity of threecerebrovascular disease.The main factors affecting the prognosis of patientswith SAH are early brain injury (EBI) and cerebral vasospasm (CVS), Somescholars believe that EBI is the most important factor affecting the prognosisof patients with SAH,EBI is the result of a variety of damage factors,eventually leading to degeneration of nerve cells, apoptosis and necrosis. Itsmechanism is complex, increased intracranial pressure, reduced cerebralblood flow, local inflammation and oxygen free radicals may be involved inthe pathogenesis, the exact mechanism has not been elucidated. it is a hotspotto research EBI mechanism in recent years. Hyperbaric oxygen therapy(HBOT) has been used in the adjuvant treatment of a variety of diseases,nevertheless, because there has pressure change in the process of hyperbaricoxygen therapy, which is rarely used to therapy early brain injury of SAH,and there is considerable controversy. In this study, we mainly observe thechange of the concentration TNF-a, S100β in the plasma and cerebrospinalfluid(CSF) of rat model of early brain injury after SAH and the change of cellapoptosis in hippocampal CA1region, and observe the change ofneurobehavioral、mortality、the concentration of TNF-a, S100β in the plasma and CSF and the cell apoptosis in hippocampal CA1region after hyperbaricoxygen therapy the EBI model rats. Thus the purpose of the study is to enrichthe theory system of EBI Occurrence and development, and to provide areference for the clinical to early use hyperbaric oxygen to therapy EBI afterSAH at the same time.Methods:96clean healthy male Sprague-Dawley rats（300-350g）wererandomly divided into normal control group (6), sham operation group (30),SAH group (30) and hyperbaric oxygenintervention SAH group (30);Thenrats of the sham-operated group, SAH group and hyperbaric oxygenintervention SAH group were further divided into five subgroups (6h,12h,24h,48h,72h) randomly according to different times of we taking thespecimens. Rats in the normal control group were fed for three days withoutany intervention and were executed to save specimens；Rats in the SAH groupand hyperbaric oxygen intervention SAH group were operated to induct aSAH model by puncturing the internal carotid artery;Rats in the hyperbaricoxygen intervention SAH group were placed in a60cm*40cm*30cm feedingcage and were intervented by HBOT at the scheduled time; Rats in thesham-operated group were operated like as SAH group, but did not puncturethe blood vessels. We recorded the death situation and neurobehavioral scoreof each group rats at the scheduled time (6h,12h,24h,48h,72h),And thenexecuting the corresponding subgroups of rats, and take the cerebrospinalfluid, brain tissue and plasma samples. we tested TNF-a, S100concentrationin the cerebrospinal fluid and plasma by enzyme-linked immunosorbent assay(ELISA),and tested cell apoptosis in hippocampal CA1area byimmunofluorescence and TUNEL.All data were recorded with Microsoft office excel2003and were analyzed with SPSS19.0. Results of measurement were presented as meanvalues±SD. Normality test and F test were performed first. Independentsample T-test,one way analysis of variance(ANOVA) and SNK-q test wereused for comparison between or among groups. The Pearson correlationcoefficient was used to assess the correlation between the level of TNF-a orS100β and the degree of cell apoptosis in the CSF and plasma;the chi-squaretest Rate was used to compared rate; the Wilcoxon rank sum test was used totest skewed data types,Statistical significance was defined as a probabilityvalue less than0.05.Result:1. The SAH group mortality rate was18.92%, the mortality rate was41.18%in the intervention group of hyperbaric oxygen, Two sets of deathsare concentrated in the early postoperative, and the hyperbaric oxygenintervention group had a higher mortality than SAH group within12hours,the mortality of two groups was statistically significant(χ2=4.897,p=0.027,p＜0.05).2. According to the amount of rats neurobehavioral score, the hyperbaricoxygen intervention group had a higher score than SAH group within12hours, and the score was lower after24hours;At the6h and12h, theneurobehavioral score of HBO intervention group was higher than the SAHgroup, which was statistically significant (p<0.05); At the24h,HBOintervention group and SAH group score was no significant difference（p>0.05）; At the48h and72h, the neurobehavioral score of HBOintervention group was lower than the SAH group, which was statisticallysignificant (p<0.05). 3. The level of TNF-a in plasma had no significant difference inSham-operated subgroups（p>0.05）,but SAH group was significantly higherthan the sham-operated group and the two groups were significantly different（p<0.001）; The level of TNF-a in plasma gradually increased in SAH group,reached to peak at24h, and then gradually decline. Compare to SAHsubgroups, the TNF-a in plasma had a higher level in hyperbaric oxygenintervention subgroups at6h and12h, and the level was lower at48h and72h. pairwise comparing two subgroups had statistical significance at6h,12h,48h and72h(p<0.05), while there was no significant difference at24h(p>0.05).4. The level of TNF-a in CSF had no significant difference inSham-operated subgroups（p>0.05）, but SAH group was significantly higherthan the sham-operated group and the two groups were significantly different（p<0.001）; The level of TNF-a in CSF gradually increased in SAH group,reached to peak at12h, and then gradually decline. Compare to SAHsubgroups, the TNF-a in CSF had a higher level in hyperbaric oxygenintervention subgroups at12h, and the level was lower at48h and72h.pairwise comparing two subgroups had statistical significance at12h,48hand72h(p<0.05), while there was no significant difference at6h and24h(p>0.05).5. The level of S100β in plasma had no significant difference inSham-operated subgroups（p>0.05）, but SAH group was significantly higherthan the sham-operated group and the two groups were significantly different（p<0.001）; The level of S100β in plasma gradually increased in SAH group,reached to peak at24h, and then gradually decline. Compare to SAHsubgroups, the TNF-a in plasma had a higher level in hyperbaric oxygen intervention subgroups at6h, and the level was lower at48h and72h.pairwise comparing two subgroups had statistical significance at6h,48h and72h(p<0.05), while there was no significant difference at12h and24h(p>0.05).6. The level of S100β in CSF had no significant difference inSham-operated subgroups（p<0.05）but SAH group was significantly higherthan the sham-operated group and the two groups were significantly different（p<0.001）; The level of S100β in CSF gradually increased in SAH group,reached to peak at12h, and then gradually decline. Compare to SAHsubgroups, the TNF-a in CSF had a higher level in hyperbaric oxygenintervention subgroups at6h and12h, and the level was lower at48h and72h.pairwise comparing two subgroups had statistical significance at6h,12h,48hand72h(p<0.05), while there was no significant difference at24h(p>0.05).7. The fluorescence intensity of cell apoptosis of hippocampal CA1areahad no significant difference in Sham-operated subgroup（sp>0.05）, but SAHgroup was significantly higher than the sham-operated group and the twogroups were significantly different（p<0.001）; The fluorescence intensityscore gradually increased in SAH group, reached to peak at12h, and thengradually decline. Compare to SAH subgroups, the fluorescence intensityscore had a higher level in hyperbaric oxygen intervention subgroups at12h,and the level was lower at48h and72h. pairwise comparing two subgroupshad statistical significance at6h,48h and72h(p<0.05), while there was nosignificant difference at12h and24h(p>0.05).8. The level of TNF-a was closely relative to the level of S100β in CSFin SAH group（r=0.867, p<0.001).9. The level of TNF-a was closely relative to the fluorescence intensity of cell apoptosis of hippocampal CA1area（r=0.729, p<0.001）.Conclusions1. The level of TNF-a in CSF in SAH rats gradually increased, reached topeak at12h, while the level of TNF-a in plasma reached to peak at24h, andthen gradually decline, but it is still high at72h; The level of TNF-a in CSFcould reflect the extent of inflammatory reaction of EBI earlier than inplasma.2. The level of S100β in CSF in SAH rats gradually increased, reached topeak at12h, while the level of S100βin plasma reached to peak at24h, andthen gradually decline, but it is still high at72h; The level of S100βin CSFcould reflect the extent of the nerve cells damage earlier than in plasma.3.The degree of hippocampal CA1neuronal apoptosis was the mostobvious at24h in subarachnoid hemorrhage model rats; Hyperbaric oxygenintervention would increase the apoptosis within12hours, while reducingapoptosis after24hours.4.The level of TNF-a in CSF in SAH rats was relative to the level ofTNF-a and the degree of hippocampal CA1neuronal apoptosis, which may beinvolved in mediating early brain injury.5.When SAH model rats were early intervented by HBO, the level ofTNF-a and S100β would increase within12hours, but it would decline after24hours.6. When SAH model rats were early intervented by HBO, it wouldaggravate the neuro cell damage within12hours, but it would alleviate injuryafter24hours. 7. When SAH model rats were early intervented by HBO,it wouldincrease the risk of death in rats within24hours, Especially within12h therewas a higher mortality rate, but it would decline after24hours. |
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