CAJ | 학술논문

目的评价硫化氢(H2S)联合浅低温对大鼠脑缺血再灌注损伤的影响.方法雄性SD大鼠80只,体重250～300 g,月龄3月,随机分为5组(n=16):假手术组(S组)、脑缺血再灌注组(IR组)、浅低温组(M组)、硫氢化钠组(NaHS组)和硫氢化钠+浅低温组(NM组).IR组、M组、NaHS组、和NM组采用四血管阻塞法建立大鼠脑缺血再灌注模型,缺血15 min后恢复灌注.NaHS组和NM组于再灌注即刻腹腔注射NaHS 14μmol/kg,其余组注射等容量生理盐水.同时M组和NM组于15 min内将直肠温降至32～33 ℃,并维持6 h;其余组采用白炽灯维持直肠温36～37 ℃.再灌注6 h时,各组处死12只大鼠,取海马组织,测定H2S的含量,采用Western b1ot法测定磷酸化cAMP反应原件结合蛋白(p-CREB)的表达,采用RT-PCR法测定脑源性神经营养因子(BDNF)mRNA的表达.于再灌注72 h时,各组处死4只大鼠,取海马组织,观察CA1区病理学结果.结果 M组、NaHS组和NM组病理学损伤较IR组减轻,其中NM组减轻最明显.与S组比较,IR组、M组、NaHS组和NM组海马H2S含量升高(P＜0.05);与IR组和M组比较,NaHS组和NM组海马H2S含量升高(P＜0.05).与S组比较,IR组、M组、NaHS组和NM组海马p-CREB和BDNF mRNA的表达上调(P＜0.05);与IR组比较,M组、NaHS组和NM组海马p-CREB和BDNF mRNA的表达上调(P＜0.05);与NaHS组和M组比较,NM组海马p-CREB表达差异无统计学意义(P＞0.05),BDNF mRNA表达上调(P＜0.05).NahS组和M组各指标比较差异无统计学意义(P＞0.05).结论 H2S联合浅低温可减轻大鼠脑缺血再灌注损伤,其机制与上调海马p-CREB和BDNF mRNA的表达有关.
목적 평개류화경(H2S)연합천저온대대서뇌결혈재관주손상적영향.방법 웅성SD대서80지,체중250～300 g,월령3월,수궤분위5조(n=16):가수술조(S조)、뇌결혈재관주조(IR조)、천저온조(M조)、류경화납조(NaHS조)화류경화납+천저온조(NM조).IR조、M조、NaHS조、화NM조채용사혈관조새법건립대서뇌결혈재관주모형,결혈15 min후회복관주.NaHS조화NM조우재관주즉각복강주사NaHS 14μmol/kg,기여조주사등용량생리염수.동시M조화NM조우15 min내장직장온강지32～33 ℃,병유지6 h;기여조채용백치등유지직장온36～37 ℃.재관주6 h시,각조처사12지대서,취해마조직,측정H2S적함량,채용Western b1ot법측정린산화cAMP반응원건결합단백(p-CREB)적표체,채용RT-PCR법측정뇌원성신경영양인자(BDNF)mRNA적표체.우재관주72 h시,각조처사4지대서,취해마조직,관찰CA1구병이학결과.결과 M조、NaHS조화NM조병이학손상교IR조감경,기중NM조감경최명현.여S조비교,IR조、M조、NaHS조화NM조해마H2S함량승고(P＜0.05);여IR조화M조비교,NaHS조화NM조해마H2S함량승고(P＜0.05).여S조비교,IR조、M조、NaHS조화NM조해마p-CREB화BDNF mRNA적표체상조(P＜0.05);여IR조비교,M조、NaHS조화NM조해마p-CREB화BDNF mRNA적표체상조(P＜0.05);여NaHS조화M조비교,NM조해마p-CREB표체차이무통계학의의(P＞0.05),BDNF mRNA표체상조(P＜0.05).NahS조화M조각지표비교차이무통계학의의(P＞0.05).결론 H2S연합천저온가감경대서뇌결혈재관주손상,기궤제여상조해마p-CREB화BDNF mRNA적표체유관.
Objective To evaluate the effect of hydrogen sulfide combined with mild hypothermia on cerebral ischemia-reperfusion (I/R) injury in rats. Methods Eighty male SD rats, aged 3 months, weighing 250-300 g, were randomly divided into 5 groups ( n = 16 each): sham operation group (group S), cerebral I/R group,mild hypothermia group (group M), sodium hydrosulfide group (group NaHS) and NaHS + mild hypothermia group (group NM). In group I/R, M, NaHS and NM, cerebral I/R was induced by occlusion of 4 vessels (cauterization of bilateral vertebral arteries and 15 min occlusion of bilateral common carotid arteries) followed by reperfusion. In group NaHS and NM, intraperitoneal NaHS 14 μmol/kg was injected immediately after reperfusion, while the equal volume of normal saline was injected in the other three groups. At the same time, the rectal temperature was reduced to 32-33 ℃ within 15 min, lasting for 6 h, in group M and NM, while it was maintained at 36-37 ℃by physical method in other groups. Twelve rats of each group were sacrificed after 6 h of reperfusion, and then the hippocampus was removed for determination of the content of H2 S by using spectrophotometer and the expression of p-CREB and BDNF mRNA by using Western blot and RT-PCR respectively. Four rats in each group were sacririced after 72 h of reperfusion and then the hippocampus was removed for microscopic examination. Results The cerebral I/R injury was attenuated in group M, NaHS and NM compared with group I/R, with the slightest injury in group NM. The H2S content was significantly higher in group I/R, M, NaHS and NM than in group S, and in group NaHS and NM than in group I/R and M. The expression of p-CREB and BNDF mRNA was significantly higher in group I/R, M, NaHS and NM than in group S, and in group M, NaHS and NM than in group I/R. The BDNF mRNA expression was significantly higher in group NM than in group M and NaHS. There was no significant difference in the H2S content and the expression of p-CREB and BNDF mRNA between group NaHS and M.Conclusion Hydrogen sulfide combined with mild hypothermia can attenuate cerebral I/R injury by up-regulating the expression of p-CREB and BDNF mRNA in hippocampus in rats.