| Objective:Hyperbaric oxygen(HBO)therapy refers to put the patients to the oxygen cabin with pressure more than one standard atmosphere and achieve the purpose of treatment by increasing the content of oxygen level in body through the breathing of high concentration oxygen with the same pressure tothe cabin,and at the same time,the treatment delays on the physical,chemical,biological and physiological effects of oxygen on body.Intracerebral hemorrhage(intracerebral hemorrhage,ICH),also known as cerebral haemorrhage,refers to the spontaneous non-traumatic intracerebral haemorrhage.Intracerebral hemorrhage(ICH)is the primary and non traumatic hemorrhage in the brain,which accounts for the acute cerebral vascular disease 20%-30%.The decompression method has become the most important emergency measures in saving the life of acuteICH patients and has been recognized by people.However,this method is still helpless in dealing with rapid improvement in preoperative and postoperative ischemia,hypoxia and cerebral edema and so on.Cerebral edema is the secondary pathological process after ICH.Reducing cerebral edema has become a key link in the clinical treatment of ICH because of the reason that effective prevention and treatment of cerebral edema is an important and effective measure to promote the recovery of neurological function and reduce the secondary brain damage.Clinical studies have indicated that HBO treatment can reduce brain edema after ICH,help the ischemic semi dark band keep alive with the improvement of blood oxygen tension and dispersion distance,and is conducive to the patient recovery,eliminate or improve dysfunction and reduce sequela.But there is still a lack of evidence of evidence-based medicine.Clinical and basic studies have shown that intracerebral hemorrhage following intervention of hyperbaric oxygen treatment can reduce brain oedema,impaired neurological function can be improved but there is no wide support from clinical and basic research data.Clinical and basic studies have shown that HBO may relieve the brain edema after the intracerebral hemorrhage and improve the neurological function impairment.But there is not have vast clinical and preclinical research date support.Aquaporins(aquaporins,AQPs)is the transport protein associated with water passthrough membrane on cell,in mammals it has been found 13 kinds of AQPs.AQP4 is the largest AQP in the brain,widely spreads in the membrane surface of astrocytes and ependymal cells of lateral basal lamina.AQP4 plays a very important role in the formation of brain edema and eliminate and physiological process,in regulating its expression in the brain has important clinical significance for the treatment of cerebral edema.Basic research shows that HBO can inhibit the expression of AQP4 in the area of intracerebral hemorrhage,and then reduce the formation of brain edema after ICH.The study found that excess free radicals can cause damage to the cell membrane,protein,mutant cell tissue induced by DNA,which caused multiplehuman diseases.The level of superoxide dismutase(SOD)in body showed a negative correlation with the free radical content.Through the detection of SOD,the content of free radicals in the body can be reflected indirectly.HBO can increase SOD(Superoxide dismutase,SOD)level,which plays a vital role on the balance between oxidation and anti-oxidation system and avoids the damage of free radicals.The previous experimental study in this research team showed that the optimal treatment for the treatment of cerebral hemorrhage in rats was 2 atmosphere absolute(ATA)pressure,and the best treated with hyperbaric oxygen in treatment of intracerebral hemorrhage is 6 hours,and inhale oxygen time is 60 min.This paper discusses the effects of different hyperbaric oxygen treatment courses on perihematomal edema and expression of aquaporin-4 and superoxide dismutase in rats with intracerebral hemorrhage,for exploring the best inhale oxygen time of hyperbaric oxygen intervene ICH.Methods:1 A total of 52 male SD rats with weight in 245g~265g,were induced to experimental intracerebral hemorrhage.These rats were offered by the Hebei Medical University laboratory animal Center,caged in the second Hospital of Hebei Medical University,Neurology,laboratory animal room,fed with standard diet and drinking tap water.The rats were divided to normal group(n=12)control group(n=20)and HBO therapy group(n=20)randomly.According to different treatment course,experimental groups and control groups was divided into different courses between 1-4 weeks,each group contains 5 rats.Experimental rats implements 1-4 weeks treatment respectively by groups.The experimental group rats were killed and retain brain tissue specimens at the end of each corresponding period after HBO therapy.Control group and normal group rats were fed in normal environment,dealed without treatment,and killed at the same time as the experimental group and made specimens.2 Made of ICH model,using the collagenase-induced method in making ICH model.3 Result evaluation,using the LongaFZ rule to judge the result 6h after model made according to the behavior change: 0 point,no sign;1 point,theopposite side of operation(the left sidebody)body can't be fully extended;2 points,turn around to the opposite side of operation,the body is limb paralysis,and appears collision phenomenon;3 points,dump to the opposite side of operation,can't stand;4 points,conscious disturbance.The model is considered success when gaining 2 points or more.4 Hyperbaric oxygen in postoperative 6h to treat when scoring to determine the successful model.Placed animal to special wooden placewith high-pressure oxygen,pressure set for 2.0ATA,pressurized 30 min,before stable time 5 minutes used straight row type to animal box filling oxygen,oxygen flow set for 10L/min(stable condition measuring oxygen concentration can reach up to 85%),stable oxygen time60 min,then decompression 20 min,1 times/d.According tothe experimentgroups,the rats were dealed with different treatment courses(1week,2week,3week,4week).5 Appraising method: AQP4 determination: Rats in each group were killed at corresponding time and took the brain with surface puncture point coronal incision,and then 3mm thick brain tissue was made to make paraffin sections.The expression of AQP4 was observed by immune-histochemistry,and cells with membrane or cytoplasm brown were determined as positive cells.Full automatic image analysis system(Image Pro Plus 6.0)was used to measure the absorbance(10*40 times)of AQP4 in the field of view.2 brain tissue sections were selected from each rat,each slice was selected for 5 different horizons,and the average light density(OD)of each group was calculated.SOD determination: Rats in each group were killed at corresponding time,took the brain tissue around the hematoma,put-80 C refrigerator preservation after liquid nitrogen cooling,and then batch determination results.When weighing,0.86% physiological saline was prepared,and the ultrasonic crushing apparatus to crush the cells,and then centrifugal 5-8min(4000r/min),took the upper part.,according to the use of manual to detection absorbance(OD).Results:1 The change of water content of rat brain tissue: results of water content around the focal cerebral hemorrhage: Comparing with the ICH control group,brain water content reached peak at 2 weeksafter ICH,and declined thereafter.1week and 2/3week control group have significantdifferent(P<0.05);no significantdifferent were found between 1week control group and 4 week control(P>0.05);significant difference was also found between 2week and 3week control group(P<0.05).comparing with corresponding control group,rats in experimental group showed significant difference and lower level brain water content(P<0.05).No significant difference were showed between 1week,3week and 4week HBOexperimental group(P>0.05),thus it can be seen,reduce of water content in brain tissueappeared earlier in experimental group than in control group.See table 1.2 The change of AQP4 expression in rat brain:AQP4 expression in control group began to rise 1 week after ICH and reach peak at 2 weeks,and gradually decreased until the fourth weeks.Significant difference were showed between each control group(P<0.05).Comparing with control group,AQP4 expression declined in one to three week experimental group and showed significant difference(P<0.05),but no significant difference were showed between 4 week experimental group and control group(P>0.05).Compare between experimental group: the trends are roughly the same as control group,but overalllower than that.AQP4 expression began to decline in 2 week group,and gradually decreased to the lowest until the fourth weeks.The experimental group appearedAQP4 expression decline earlier and larger range than control group.No significant difference were showed between 1 week and 3week group,but significant difference were showed respectively between 2 week and 4 week group(P<0.05),and significant difference were also showed between 2 week and 4 week group(P<0.05).See table 2.3 The change of SOD expression in rat brain: T-SOD:SODexpression of experimental group with HBO treatment reach the peak in 2 weeks after ICH,and then gradually decreased.The control group and the experimental group have the same trend on SODexpression.Significant difference were showed between 4 and 1,2,3 week experimental group(P<0.05).No significant difference were showed between 1week to 3week experimental group(P<0.05).According to the data,significant difference were showed between 4 and 1,2,3 week control group(P<0.05).No significant difference were showed between 1week to 3week control group(P<0.05).Significant difference were showed between each experimental group and control group(P<0.05).Cu-ZnSOD:compare of SOD expression between control group andexperimental group with HBO treatment,no significant difference wereshowed between each group in experimental groups(P>0.05).No significantdifference were showed between each control group(P>0.05).Significantdifference were showed between1 week,2week and 4week in experimentalgroup and control group(P<0.05).No significant difference were showedbetween3week in experimental group and control group(P>0.05).Conclusion:This paper studied the treatment course of hyperbaric oxygen through the establishment of rat model of cerebral hemorrhage.Under the condition: 6h after ICH,2.0 ATA pressure of HBO,inhale oxygen time 60 min,and the period 1time per day,7 times per week.After ICH,the changes of AQP4 and SOD expression in rats with different treatment of cerebral hemorrhage after HBO treatment showed that the change of cerebral water content was consistent with the expression of AQP4 in rats.The study found the expression of AQP4 increased in the control group after first weeks of ICH,reached a peak of second weeks,decreased gradually in third week,and restored to the normal leve in the fourth week.Compared with the control group,the experimental group was lower than the control group.The study of SOD change found the expression of SOD in control groups increased in the first three weeks,reached peak in second week,and decreased to the normal leve in the fourth week.The expression of the experimental group was increased comparing with the control group.Thus,HBO can reduce brain edema after ICH,increase antioxidant capacity,and treatment for 4 weeks has better effects than other courses. |
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