高渗盐溶液干预对心肺复苏后小型猪的保护作用及机制的实验研究

发布时间：2018-04-11 12:18

本文选题：心脏骤停 + 心肺复苏　；参考：《北京协和医学院》2016年博士论文

【摘要】：目的：研究高渗盐溶液对心肺复苏后小型猪的保护作用及可能的机制。方法：27只小型猪随机分配至3个组:GroupI(n=9)---健康小型猪诱颤后3min静注生理盐水(Normal saline, NS)4ml/kg; GroupII(n=9)---健康小型猪诱颤后3min静注7.5%高渗盐溶液(Hypertonic saline, HS)4ml/kg; GroupⅢ(n=9)—-健康小型猪诱颤前建立全身炎症反应综合征(Systemic inflammatory response syndrome, SIRS)模型,诱颤后3min静注7.5%HS4ml/kg。所有实验对象在诱颤后3min开始心肺复苏(Cardiopulmonary Resuscitation, CPR),记录各组自主循环恢复(return of spontanesou circulation, ROSC)率,ROSC后120min存活率；基础及ROSC即刻、30、60、90、120min相关血流动力学指标；基础及ROSC即刻、60、120min血清炎症指标。每组动物ROSC后120min处死并取心脑肾肠,测定组织中的髓过氧化物酶(Myeloperoxidase, MPO),丙二醛(Malondialdehyde, MDA),超氧化物歧化酶(Superoxide dismutase, SOD)浓度,光镜下对组织器官病理形态进行观察,末端脱氧核苷酸转移酶末端标记法(Terminal-deoxynucleotidyl tra nsferase mediated nick end labeling, TUNEL)检测各器官细胞凋亡指数,链霉亲和素-生物素-酶复合物(Strept Avidin-Biotin Complex, SABC)法免疫组化检测心肌组织Bcl-2、Bax、FasL蛋白表达。结果：1.组间ROSC率、120min存活率无统计学差异,但G3组CPR时间长于G2组(P0.05)。2.G3组ROSC后心率、中心静脉压高于其余两组(P0.05),G2组只在ROSC后30min心率高于G1组(P0.05)；G1组及G2组ROSC后平均动脉压、冠脉灌注压、颈动脉血流速无统计学差异,G3组ROSC后60min开始冠脉灌注压低于其他两组(P0.05),ROSC后90min开始平均动脉压低于其余两组(P0.05),ROSC后60及120min颈动脉血流速低于G2组(P0.05)。3.实验对象血清炎症介质在ROSC后均显著升高,G3组血清炎症介质升高更为明显；G2组血清TNF-α在ROSC后60及120min均低于G1组(P0.05),IL-1β只在ROSC后60min低于G1组(P0.05)；G1,1间组血清IL-4、IL-10同一时间点无统计学差异。4.G2组心肌组织SOD活性高于其余两组(P0.05),G1,3组间无统计学差异,G1,2MDA及MPO活性低于G3组(P0.05),G1组和G2组组间无统计学差异；G2,3组脑组织SOD活性高于G1组(P0.05),MDA活性低于G1组(P0.05),但组间无统计学差异,三组间MPO活性无统计学差异；三组间小肠组织SOD活性无统计学差异,G3组MDA及MPO活性高于其余两组(P0.05),G1,2组间MDA、MPO活性无统计学差异；三组间肾脏组织SOD、MDA活性无统计学差异,G3组MPO活性高于其余两组(P0.05),G1,2组间无统计学差异。5.G3组心脑组织病理损伤评分高于其余两组(P0.05),G2组心脏病理损伤评分低于G1组(P0.05),但G1,2组脑病理损伤评分无统计学差异；三组间小肠病理损伤评分无统计学差异；G3组肾脏病理损伤评分高于G2组(P0.05),G1组与其余两组无统计学差异。6.G2组心肌细胞凋亡低于其余两组(P0.05),G1,3组无统计学差异；G3组脑、肠、肾组织细胞凋亡均高于其余两组(P0.05),G1,2组无统计学差异,但G2组脑细胞凋亡有减少趋势。7.G1组有少量Bcl-2、Bax及FasL蛋白表达;G2组可见Bcl-2蛋白表达增高,FasL蛋白表达降低,与G1组比较均有统计学差异(P0.05),而Bax表达无统计学差异；G3组Bcl-2蛋白表达低于G2组(P0.05),Bax及FasL蛋白表达指数很高,与其他两组比较均有统计学差异(P0.05)。结论：1.HS溶液干预对CPR后小型猪ROSC率、存活率、平均动脉压、冠脉灌注压、颈动脉血流速无明显影响,但可一过性提高中心静脉压。2.HS抑制炎症介质释放,对抑炎介质的生成可能无影响。3.HS促进心脑组织SOD生成,对小肠、肾组织缺乏类似作用,同时降低脑组织MDA浸润。4.HS减轻缺血后心肌损伤,抑制心肌细胞凋亡发生,有降低脑细胞凋亡趋势。5.HS下调促凋亡基因FasL蛋白表达、上调抑凋亡基因Bcl-2蛋白表达,可能是其抗心肌细胞凋亡的分子机制的重要环节。
[Abstract]:Objective: To study the protective effect of hypertonic saline solution on cardiopulmonary resuscitation in pigs and its possible mechanism. Methods: 27 pigs were randomly assigned to 3 groups: GroupI (n=9) - induced VF healthy pigs 3min after intravenous injection of saline (Normal, saline, NS) 4ml/kg; GroupII (n=9) - Health small pig induced fibrillation after 3min intravenous injection of 7.5% hypertonic saline solution (Hypertonic saline, HS 4ml/kg); Group III (n=9) - induced VF healthy pigs before the establishment of systemic inflammatory response syndrome (Systemic inflammatory response syndrome, SIRS) model, 3min induced fibrillation after intravenous injection of 7.5%HS4ml/kg. in all subjects after 3min induced atrial fibrillation CPR (Cardiopulmonary Resuscitation, CPR), recorded the restoration of spontaneous circulation (return of spontanesou circulation, ROSC ROSC) rate, the survival rate of 120min; and based at ROSC, 30,60,90120min and ROS based hemodynamic index; At C, serum inflammatory markers after ROSC 120min 60120min. Each animal were sacrificed and cardiovascular and renal tissue in the intestine, determination of myeloperoxidase (Myeloperoxidase, MPO), malondialdehyde (Malondialdehyde, MDA), superoxide dismutase (Superoxide dismutase, SOD) concentration, light microscopy of tissue and organ pathological morphology were observed. The end DEOXYNUCLEOTIDYLTRANSFERASE endlabeling (Terminal-deoxynucleotidyl tra nsferase mediated nick end labeling, TUNEL) to detect the organ cell apoptosis index, streptavidin biotin peroxidase complex (Strept Avidin-Biotin, Complex, SABC) immunohistochemical detection of myocardial Bcl-2, Bax, FasL protein expression between the 1. groups. Results: the rate of ROSC 120min, the survival rate was no significant difference, but the G3 group CPR group G2 group.2.G3 (P0.05) ROSC heart rate, central venous pressure is higher than the other two groups (P0.05), G2 group 30min after ROSC The heart rate is higher than that of G1 group (P0.05); G1 group and G2 group after ROSC mean arterial pressure, coronary perfusion pressure, there was no significant difference in carotid artery blood flow, 60min G3 group ROSC after coronary perfusion pressure is lower than the other two groups (P0.05, 90min) to mean arterial pressure after ROSC lower than the other two groups (P0.05, ROSC) after 60 120min and carotid artery blood flow velocity is lower than that of group G2 (P0.05) serum inflammatory mediators.3. subjects were significantly increased in ROSC, G3 group of serum inflammatory mediators were significantly increased in G2 group; serum TNF- after ROSC 60 and 120min were lower than those in group G1 (P0.05), IL-1 only after ROSC 60min was lower than that of beta G1 group (P0.05); G1,1 group of serum IL-4, IL-10 at the same time no SOD activity in myocardial tissue of.4.G2 group significant difference than the other two groups (P0.05), no significant difference between groups G1,3, G1,2MDA and MPO were lower than group G3 (P0.05), G1 group and G2 group had no statistical difference; the activity of SOD in brain tissue of G2,3 group Higher than that of G1 group (P0.05), the activity of MDA was lower than that of G1 group (P0.05), without significant difference between groups, no significant difference between the three groups MPO activity; no significant difference between the three groups of small intestine tissue SOD activity, MDA and MPO activity in G3 group than that in the other two groups (P0.05), G1,2 group, MDA, MPO. No statistically significant difference between the three groups; kidney tissue SOD, there was no significant difference in the activity of MDA, G3 in MPO group was higher than the other two groups (P0.05), no pathological damage of heart and brain of.5.G3 group were higher than those of the other two groups statistically significant difference between the G1,2 group (P0.05), G2 group cardiac pathology score was lower than that of G1 group (P0.05). But the group of brain injury G1,2 score had no significant difference; small intestine pathological damage score was no significant difference between the three groups; G3 group renal pathological injury score higher than that of G2 group (P0.05), G1 group and the other two groups had no statistical difference of myocardial apoptosis in.6.G2 group is lower than the other two groups (P0.05), no significant group G1,3 The difference in G3 group; brain, intestine, renal cell apoptosis were higher than those of the other two groups (P0.05), no significant difference in the G1,2 group, but the G2 group had a tendency to decrease the apoptosis of brain cells in.7.G1 group had a small amount of Bcl-2, the expression of Bax and FasL increased; G2 group showed the expression of Bcl-2 protein, FasL protein expression decreased, compared with the G1 group had significant difference (P0.05), and the expression of Bax had no significant difference; the expression of Bcl-2 protein in G3 group was lower than that of G2 group (P0.05), Bax protein and FasL expression index is very high, and the other two groups were statistically significant (P0.05). Conclusion: 1.HS solution intervention on the survival rate of ROSC miniature pig rate, CPR, mean arterial pressure, coronary perfusion pressure, carotid artery blood flow had no significant effect, but can improve a central venous pressure.2.HS inhibited the release of inflammatory mediators, to produce anti-inflammatory medium may promote the formation of SOD.3.HS has no effect on heart and brain, small intestine, kidney deficiency with similar effects. To reduce myocardial injury of brain tissue infiltration of MDA.4.HS reduced after ischemia, inhibiting myocardial apoptosis, decrease the apoptosis of brain cells.5.HS protein expression down-regulation of the pro apoptotic gene FasL, up-regulated expression of anti apoptosis gene Bcl-2 protein may play an important role in the anti apoptosis mechanism.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R459.7

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