Study On The Influence Of Hyperbaric Oxygen Therapy To Expression Of P53and Caspase-3in Early Brain Injury After Subarachnoid Hemorrhage

Objective: Subarachnoid hemorrhage (SAH) is one of the high mortalityand high disability rate of cerebral vascular disease, it divided intospontaneous and traumatic. Accounted for all of the spontaneoussubarachnoid hemorrhage80%cause of subarachnoid hemorrhage of rupturedintracranial aneurysm, also called the aneurysmal subarachnoid hemorrhage,the death rate and disability rate is very high, the mortality was45%within3days after the hemorrhage. Scholars believe that early brain injury (EBI) afterhemorrhage is one of the main factors of high mortality and disability rate.Hyperbaric oxygen therapy (HBOT) is inhalation of pure oxygen at higherthan an atmospheric environment, it has been widely used in the treatment ofmany diseases presently, due to the SAH patients condition changes quicklyand high risk, few studies of early hyperbaric oxygen therapy for SAH. Thisstudy established the rat model of SAH by internal carotid artery puncture,observes the expression of P53and Caspase-3in cerebral cortex of rats, andthe changes of neurobehavior, mortality, expression of P53and Caspase-3incerebral cortex after HBOT.Methods:100healthy adult male Sprague-Dawley (SD) rats that weightof280g to320g, they were randomly divided into control group (10), shamoperation group (30), SAH group (30) and hyperbaric oxygen treat SAHgroup (30); In addition to the control group, the rest groups were divided intothree subgroups (24h,48h,72h) by the experimental animal different specimen time, each subgroup10rats. The blank control group without anyspecial treatment, anesthesia, perfusion, take brain tissue samples afterconventional feeding3days; SAH group and HBOT+SAH group rats wereused puncturing the internal carotid artery endovascular to make SAH model,except HBOT+SAH group rats were treated with hyperbaric oxygen after thesuccess of modeling, the rearing environment and postoperative care were allthe same; the modeling process of sham group is same as the SAH group, butdo not puncture vessel. Record each subgroup neurobehavioral scores andmortality of sham group, SAH group and HBOT+SAH group at24h,48h and72h after modeling; then, anesthesia and perfusion each subgroup rats, takethe brain tissue preservation. Immunohistochemical method was used todetect the expression of P53and Caspase-3in cerebral cortex.Using Motic Image Advance3.2image analysis system to analyze theimmune tissue sections, calculate the percentage of positive cells, all the datarecord by Microsoft office excel2003, normal distribution measurement datais represented by S. Data were analyzed by spss19.0software, normalitytest and F test were performed first, two groups were compared using the ttest, comparison among subgroups by SNK method (q test), P <0.05wasconsidered statistically significant.Result:1. Mortality: The mortality rate of blank control group and shamoperation group rats were0, The two groups had no statistical significance.The total mortality rate in SAH group and HBOT+SAH group were40%and16.67%, which was statistically significant (P <0.05); The mortality rates ofSAH group and HBOT+SAH group were23.33%and13.33%in24hours,the mortality within24hours of the two groups were no significant difference (P＞0.05).2. Neurobehavioral score: It had no significant differences that comparedbetween SAH group and HBOT+SAH group at24h and48h; But the score ofHBOT+SAH group was significantly lower than that in SAH group at72h,the difference was statistically significant (P <0.05); In SAH group, it had nosignificant difference between the each two subgroup (P＞0.05); Comparedbetween the three subgroup of HBOT+SAH group: it was no statisticallysignificant between24h and48h (P＞0.05), but they have statisticallysignificant between24h and72h,48h and72h (P <0.05).3. The positive expression of P53in cerebral cortex is brown or brownyellow particles in the nucleus, and a small amount of expressed in cytoplasm.Blank control group and sham operation group were lower expression, thetwo groups had no statistical significance (P＞0.05). Compared between thethree subgroup of HBOT+SAH group: It is only between24h and48h that theexpression of P53has no difference, each SAH subgroup have moresignificantly positive expression than each sham subgroup at every time point,the difference have statistical significance (P <0.05); it have statisticalsignificance between the three subgroup of HBOT+SAH group; Theexpression of P53in HBOT+SAH group was lower than that of SAH group at48h and72h, the difference have statistical significance (P <0.05);HBOT+SAH group and SAH group has no difference at24h (P＞0.05).4. Positive expression of Caspase-3is brown granules in the cytoplasm,Blank control group and sham operation group were lower expression, thetwo groups had no statistical significance (P＞0.05). Each SAH subgrouphave more significantly positive expression than each sham subgroup at everytime point, the difference have statistical significance (P <0.05); Difference of the three subgroup in SAH group have statistical significance, also as theHBOT+SAH group. Compared the expression of Caspase-3between SAHgroup and HBOT+SAH group, all the difference have statistical significanceat24h,48h and72h (P <0.05).Conclusions:1. Early brain injury after SAH is related to the increase of expression ofP53and Caspase-3in rats cerebral cortex.2. HBOT can reduce the total mortality of rats in the early days (in72h)after SAH, and improve the neurological function in a certain extent, haveeffect on the treatment of EBI, but to reduce the mortality rate of SAH rats in24h is not obvious.3. Hyperbaric oxygen may be through decreasing the expression of P53and Caspase-3to protect brain tissue in the early after SAH.