CAJ | 학술논문

目的评价富氢液对大鼠短暂性脑缺血再灌注时炎性反应的影响.方法健康雄性SD大鼠45只,5～6周龄,体重180～ 250 g,采用随机数字表法,将其分为3组(n=15):假手术组(S组)、脑缺血再灌注组(I/R组)和富氢液组(H组).采用结扎双侧颈总动脉15 min伴低血压-再灌注的方法建立大鼠短暂性脑缺血再灌注损伤模型.H组于再灌注即刻腹腔注射0.6 mmol/L富氢液5 ml/kg;I/R组再灌注即刻腹腔注射等容量生理盐水.于造模前5d开始,每组取5只大鼠,采用Morris水迷宫法测定认知功能,定位航行实验历时5d,记录逃避潜伏期、游泳速度以及游泳总路程;造模后1和3d时行空间探索实验,记录目标象限停留时间百分比、穿越平台次数和游泳速度.造模后3d空间探索实验结束后,处死大鼠,取海马组织,光镜下观察海马CA1区神经元形态.于造模后1d时,每组处死5只大鼠,取海马组织,采用ELISA法检测TNF-α和IL-1β的含量;每组处死5只大鼠,取海马组织,采用免疫组化法测定NF-κB活性.结果造模前定位航行实验中,3组游逃避潜伏期和游泳路程随训练时间的延长逐渐缩短(P＜0.05),游泳速度随训练时间的延长无改变(P＞0.05).与S组比较,I/R组造模后1和3d时探索实验中目标象限停留时间百分比降低,穿越平台次数减少,造模后1d时海马组织TNF-α和IL-1β的含量、NF-κB活性升高(P＜0.05).与I/R组,H组造模后1和3d时探索实验中目标象限停留时间百分比升高,穿越平台次数增加,造模后1d时海马组织TNF-α和IL-1β的含量、NF-κB活性降低(P＜0.05),病理学损伤减轻.3组间造模后1和3d时探索实验中游泳速度比较差异无统计学意义(P＞0.05).结论富氢液减轻大鼠短暂性脑缺血再灌注损伤的机制可能与其抑制炎性反应有关.
목적 평개부경액대대서단잠성뇌결혈재관주시염성반응적영향.방법 건강웅성SD대서45지,5～6주령,체중180～ 250 g,채용수궤수자표법,장기분위3조(n=15):가수술조(S조)、뇌결혈재관주조(I/R조)화부경액조(H조).채용결찰쌍측경총동맥15 min반저혈압-재관주적방법건립대서단잠성뇌결혈재관주손상모형.H조우재관주즉각복강주사0.6 mmol/L부경액5 ml/kg;I/R조재관주즉각복강주사등용량생리염수.우조모전5d개시,매조취5지대서,채용Morris수미궁법측정인지공능,정위항행실험력시5d,기록도피잠복기、유영속도이급유영총로정;조모후1화3d시행공간탐색실험,기록목표상한정류시간백분비、천월평태차수화유영속도.조모후3d공간탐색실험결속후,처사대서,취해마조직,광경하관찰해마CA1구신경원형태.우조모후1d시,매조처사5지대서,취해마조직,채용ELISA법검측TNF-α화IL-1β적함량;매조처사5지대서,취해마조직,채용면역조화법측정NF-κB활성.결과 조모전정위항행실험중,3조유도피잠복기화유영로정수훈련시간적연장축점축단(P＜0.05),유영속도수훈련시간적연장무개변(P＞0.05).여S조비교,I/R조조모후1화3d시탐색실험중목표상한정류시간백분비강저,천월평태차수감소,조모후1d시해마조직TNF-α화IL-1β적함량、NF-κB활성승고(P＜0.05).여I/R조,H조조모후1화3d시탐색실험중목표상한정류시간백분비승고,천월평태차수증가,조모후1d시해마조직TNF-α화IL-1β적함량、NF-κB활성강저(P＜0.05),병이학손상감경.3조간조모후1화3d시탐색실험중유영속도비교차이무통계학의의(P＞0.05).결론 부경액감경대서단잠성뇌결혈재관주손상적궤제가능여기억제염성반응유관.
Objective To evaluate the effect of hydrogen-rich saline on inflammatory responses during transient cerebral ischemia-reperfusion (I/R) in rats.Methods Forty-five male Sprague-Dawley rats,aged 5-6 yr,weighing 180-250 g,were randomly divided into 3 groups (n =15 each) using a random number table:sham operation group (group S),group I/R and hydrogen-rich saline group (group H).The rats were anesthetized with intraperitoneal 10％ chloral hydrate 0.3 ml/100 g.Transient cerebral ischemia was induced by bilateral common carotid artery occlusion with hypotension for 15 min,followed by reperfusion.Five rats were randomly chosen from each group,and Morris water maze was used to assess the cognitive function starting from 5 days before establishment of the model.Place navigation test lasted for 5 consecutive days.The escape latency,swimming speed and swimming distance were recorded.Spatial probe test was carried out on 1 and 3 days after establishment of the model.The time of staying at the target platform quadrant,frequency of crossing the original platform and swimming speed were recorded.The rats were sacrificed after the end of spatial probe test on 3 days after the model was established,and hippocampi were removed to examine the morphology of neurons in hippocampal CA1 region with light microscope.Five rats randomly chosen from each group were sacrificed on 1 day after the model was established,and hippocampi were removed to detect the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β).The rest 5 rats in each group were sacrificed,and hippocampi were removed for determination of nuclear factor kappa B (NF-κB) activity (by immuno-histochemistry).Results In place navigation test before the model was established,the escape latency and swimming distance were gradually shortened with the prolonging training time,and no significant change was found in the swimming speed with the prolonging training time in the three groups.Compared with group S,the time of staying at the target platform quadrant was significantly shortened,and the frequency of crossing the original platform was reduced on 1 and 3 days after establishment of the model,and the contents of TNF-α and IL-1β and NF-κB activity were increased on 1 day after establishment of the model in group I/R.Compared with group I/R,the time of staying at the target platform quadrant was significantly prolonged,and the frequency of crossing the original platform was increased on 1 and 3 days after establishment of the model,and the contents of TNF-α and IL-1β and NF-κB activity were decreased on 1 day after establishment of the model in group H.There was no significant change in the swimming speed during spatial probe test on 1 and 3 days after establishment of the model.Conclusion The mechanism by which hydrogen-rich saline reduces transient cerebral I/R injury may be related to inhibition of inflammatory responses in rats.