【作者】 郭涛；

【导师】 黄亮；

【作者基本信息】 南昌大学 ， 急诊医学， 2011， 硕士

【摘要】 目的：探讨H2S在心肺复苏后脑损伤中的作用。方法：取SD雄性大鼠45只,随机分为假手术(A)组、CPR(B)组和NaHS处理(C)组,A组5只,B组、C组按自主循环恢复(restoration of spontaneous circulation,ROSC)后6h、12h、24h、72h四个时间点各分四个亚组,每亚组各5只,每组共20只。参照经典窒息法制作心脏骤停模型。A组仅行置管及插管,B组制作心肺复苏模型大鼠,C组在B组基础上于CPR前一分钟静脉推注NaHS 10umol/kg(将称取的NaHS立即溶于0.5m1生理盐水中)。采用全自动酶标仪测定大鼠脑皮质H2S含量；721分光光度计测脑组织EB含量；免疫组化计Nestin阳性细胞数；测脑组织干/湿重比并采用原子吸收光谱法测脑组织钠、钾离子含量,行NDS评分。结果：(1)脑组织H2S含量变化：与A组相比,B组6h、12h脑组织H2S含量增加(6h,p<0.05,12h,p<0.01),24h下降(p<0.05),72 h恢复A组水平(p>0.05)。与B组同时间点比,C组中各时间点脑组织H2S含量增加(p<0.05)。(2)脑水肿指标变化：与A组相比,B组脑组织含水量、EB含量、钠离子含量随时间点逐渐上升,至24h达到高峰,72h呈下降趋势。钾离子含量逐渐下降,24h至低谷,72h呈上升趋势,与B组同时间点比,C组6h、12h、24 h EB含量下降(p<0.05),C组12h、24 h、72 h脑组织含水量及脑组织钠离子含量下降,钾离子含量上升(p<0.05)。(3)脑组织巢蛋白、NDS评分变化：与A组相比,B组巢蛋白、NDS评分表达升高,且其值随时间点逐渐增加(p值均<0.01),与B组同时间点比,C组各时间点巢蛋白表达、NDS评分增加(p值均<0.05)。行相关性分析显示：脑组织含水量与脑组织钠离子含量变化呈正相关(r=0.688,p<0.01),与脑组织钾离子含量呈负相关(r=-0.554,p<0.05)；与脑组织EB含量相关(r=0.854,p<0.01),将H2S与脑组织含水量进行相关性分析,结果显示B组中,H2S与脑组织含水量不相关,C组H2S与脑组织含水量呈负相关(r=-0.663,p<0.05)。结论：小剂量外源性H2S可通过降低大鼠脑组织钠离子含量,增加钾离子含量,减轻BBB通透性的增加而降低CPR后脑组织含水量,同时还可上调脑组织巢蛋白的表达,改善NDS评分；H2S可能通过上述机制影响心肺复苏后脑损伤的发生发展。更多还原

【Abstract】 Objective:To investigate the role of H2S in the formation of brain edema and the effect of H2S on the expression of nestin in hippocampus after CPR.Methods:Forty-five male Sprague-Dawley(SD) rats were randomly diveded into normal group(group A), model group(group B) and NaHS pretreatment group(group C). There were 5 rats in sham-operated group,20 rats in model group and NaHS pretreatment group,20rats were separately divided into four subgroups at 6h、12h、24h、72h after ROSC with 5 rats in each subgroup.The cardiac arrest model of rat was established according to asphyxy method. Catheters were inserted into left femoral artery and vein of rats while tracheal intubation was performed in A group, the cardiac arrest model of rat was established in rats of group B and C,0.5ml NaHS solution (lOumol/kg NaHS dissolving in physiological saline by the rat weight) was administered intravenously 1 minute before CPR. The level of H2S in cortex in brain tissue of rats in three groups were determined by universal Microplate Reader, the content of EB were measured by 721 spectrophotometer. immunohistochemistry method was adopted to calculate the amount of Nestin positive cells, the ratio of wet to BWC was calculated, atomic absorption spectrometry was adopted to measure the content of Na+and K+, NDS score was calculated.Results:(1) Change of the level of H2S in brain tissue:Compared with group A, the level of H2S in brain tissue increased at 6h、12h、(6h、p<0.05,12h、p<0.01), and decreased at 24h (p<0.05),assumed a dynamic chang of rising and descending, returning to group A level at 72h (p>0.05). Compared with group B, the level of H2S in all time points of group C increased (p<0.05) in brain tissue. (2) Change of indicator in celebral edema:Compared with group A, the content of water, EB and Na+ of brain tissue increased gradually, peaked at 24h and assumed a trend of decreasing at 72h, the content of K+ of brain tissue decreased gradually, lowered at 24h and assumed a trend of increasing at 72h, Compared with crresponding time point of group B, the content of EB was decreased at 6h、12h、24 h in C group (p<0.05),the content of water and Na +decreased while the content of K+increased at 12h、24 h、72h in C group (p<0.05). (3)Change of the expression of nestin and NDS score:Compared with group A, the expression of nestin and NDS score increased in B group (p<0.01) and its data increased gradually with time points. Compared with crresponding time point of group B, the expression of nestin and NDS score increased in all subgroups of group C (p<0.05), Correlation analyse demonstrated that the content of water of brain tissue was positively correlated with the content of Na+ of brain tissue (r=0.688, p<0.01), negatively correlated with the content of K+ of brain tissue (r=-0.686, p<0.01), and was positively correlated with the content of EB of brain tissue (r=0.854,p<0.01);the level of H2S wasn’t correlated with brain edema in group B, and was positively correlated with brain edema in group C (r=-0.663,p<0.01)Conclusion:low level of exdogenous Hydrogen sulphide can decrease brain water content by decreasing the content of Na+ in brain tissue、increasing the content of K+ in brain tissue and mitigating BBB Permeability, while upregulate the expression of nestin in brain tissue,and thus increase NDS score. H2S may influence in the formation and progression of celebral injury after CPR though the above mechanism,更多还原