不同心肌保护方法对体外循环下胎羊心功能的影响

发布时间：2018-06-25 10:10

本文选题：STH液 + HTK液　；参考：《广西医科大学》2017年硕士论文

【摘要】：目的通过观察比较St.Thomas停搏液(STH液)、组氨酸-色氨酸-酮戊二酸停搏液(HTK液)和不停跳对胎羊实行体外循环(CPB)术中及术后心肌酶、心肌组织结构、代谢及实验中其它相关指标的影响,评估这三种方法对胎羊心肌的保护作用,寻求一种适合的胎羊体外循环心肌的保护方法,从而为临床胎儿体外循环提供依据。方法18只怀孕山羊(孕100—120 d,足月为150 d),重量为25.72±4.29 kg(17.5-32 kg),共孕有18只胎羊。分为3组:不停跳组(不停跳组),STH组(St.Thomas保护液组),HTK组(HTK保护液组)。3组胎羊均采用胸骨前正中切口,完全肝素化后建立CPB行主肺动脉-右心房插管转流,体外循环回路建立好后开始体外循环,其中St.Thomas保护液组和HTK保护液组在体外循环开始后,用主动脉钳夹闭主动脉开始灌流心肌保护液,不停跳组不灌流心肌保护液。控制胎羊体外循环流量在200 ml/kg/min以上,水箱温度恒定40℃。St.Thomas保护液组和HTK保护液组使胎羊心脏停跳30分钟,然后开放升主动脉在并行循环下使胎羊心脏复跳,30分钟后撤离体外循环,观察2小时,不停跳组转流60 min后停机观察2h。在CPB前、转流30min、转流60min、停机后1h、停机后2h采颈静脉血5ml,腋动脉动脉血1ml(测血气)。胎羊静脉血均在4℃下4000 rpm离心15min,取上清液,分装入EP管(每管约0.5ml),放入-80℃冰箱冻存。在停机后2h,胎羊静脉注射10%氯化钾5 ml,使心跳迅速停跳,剪取胎羊右心房心肌组织大小约1×1×1cm,置入含有4%福尔马林液的试管中密封保存于常温中,待1周后进行脱水、石蜡包埋,动物实验完成后用TUNEL统一染色观察细胞凋亡情况,剪取心室肌若干块,大小约0.5×0.5×0.5cm用于乳酸和ATP的检测。分别于转流前(T1)、转流30 min(T2)、转流60 min(T3)、转流结束后1h(T4)、转流结束后2 h(T5)测心率(Heart Rate,HR)、平均动脉压(Mean Artery Pressure,MAP)、体外循环转流量、PH值、PaO_2、Pa CO_2、碱剩余(Base Excess,BE)、K~+、Na~+、Ca~(2+)、血糖(Glucose,Glu)、乳酸(Lactic Acid,Lac)、血红蛋白(Hemoglobin,Hb),在各个时间点采集胎羊静脉血检验心肌受损指标:CKMB、钙蛋白I(c Tn I)和肌钙蛋白T(c Tn T)。本次研究采集的所有数据均用SPSS21.0统计学软件处理,计量资料以均数±标准差(x±s)的方法表示。两组间差异显著性检验利用单因素方差检验(Student’s t-test),多组间显著性比较采用one-way ANOVA检验。结果1.三组胎羊在各个时间点腋动脉在测得的血气中,p H、PO_2(mm Hg),PCO_2(mm Hg)、K~+(mmol/L)、Ca2+(mmol/L)在各时间点比较均无统计学差异(P0.05)。在T3及T3以后的时段HTK组Na+(mmol/L)要低于STH组(P0.05);在T3时,HTK组的K~+(mmo/L)、Lac(mmol/L)、BE(mmol/L)浓度低于STH组(P0.05);T4时,HTK组的Hct(%)高于STH组,有统计学差异(P0.05)。2.体外循环之前胎羊血清c Tn T、c Tn I、CKMB浓度分别在三组间比较均无明显差异(P=0.430,P=0.391,P=0.071)。血清c Tn I(ng/L)浓度变化,在T1、T2、T3时三组间比较无统计学差异(P0.05);T4时STH组的c Tn I较CPB前明显升高(P0.05);T5时不停跳组与HTK组的c Tn I均较比CPB前升高(P0.05);T4和T5时HTK组的c Tn I低于STH组(P0.05);T4时STH组和T5时的STH组、HTK组的c Tn I高于不停跳组(P0.05)。3.HTK组和STH组TUNEL阳性细胞数均较不停跳组组明显增多(p=0.001和p=0.048),但STH组TUNEL阳性细胞数较HTK组显著增多(p=0.007)。4.STH组心肌组织ATP含量较不停跳组与HTK降低(P0.05);HTK组的乳酸含量高于不停跳组(P0.05);STH组的乳酸含量高于不停跳组和HTK组(P0.05)结论胎羊体外循环中,不停跳对心肌的保护明显优于停跳。在胎羊停跳方面与STH停搏液相比,HTK液在降低心肌代谢、减少心肌损伤标记物的释放方面有明显优势,TUNEL标记的细胞凋亡减少,心肌ATP含量有增加。HTK液的停跳时间要长与STH液相比可引起低钠血症。
[Abstract]:Objective To observe and compare the effects of St.Thomas cardioplegia solution (STH solution), histidine tryptophan - ketopulacid cardioplegic solution (HTK solution) and nonstop jump on cardiac muscle enzymes, myocardial tissue structure, metabolism and other related indexes during and after extracorporeal circulation (CPB) in fetal sheep, and to evaluate the protective effect of these three methods on fetal sheep myocardium. The protective method of cardiopulmonary bypass was suitable for fetal cardiopulmonary bypass to provide basis for clinical fetal cardiopulmonary bypass. Methods 18 pregnant goats (pregnancy 100 to 120 d, full month 150 d), 25.72 + 4.29 kg (17.5-32 kg) and 18 pregnant sheep were divided into 3 groups: non stop jump group (non stop jump group), STH group (St.Thomas protection solution group), HTK group (HTK protection solution group). The 3 groups of fetal sheep were all used in the anterior sternum incision. After completely heparin, CPB was set up with the main pulmonary artery right atrium intubation. After the cardiopulmonary bypass was established, the St.Thomas protective fluid group and the HTK protection group began to use the aorta clamp closed aorta after the cardiopulmonary bypass. The flow of cardiopulmonary bypass was controlled by the cardiopulmonary bypass. The flow rate of the fetal sheep was over 200 ml/kg/min, the temperature of the water tank was constant at 40 centigrade and the HTK protection solution group made the fetal heart jump for 30 minutes. Then the ascending aorta was opened in the parallel circulation to make the fetal heart jump, 30 minutes after the evacuation of the cardiopulmonary bypass, for 2 hours and 60 MI in the non stop group. After n, 2h. was stopped to observe the flow of 30min, transfer of 30min, transfer 60min, 1h after shutdown, 2h take neck vein blood 5ml, and the arterial blood of axillary artery is 1ml (blood gas). The venous blood of fetal sheep is centrifuged under 4 centigrade, and the supernatant is loaded into the EP tube (each tube about) and stored in the refrigerator. After the shutdown, 10% potassium chloride 5 of fetal sheep are injected, so that the fetal sheep vein is injected intravenously. The cardiac muscle tissue of the right atrium was reduced to 1 x 1 x 1cm in the right atrium of the fetal sheep. The test tube containing 4% formalin liquid was sealed in the normal temperature, and after 1 weeks, dehydration and paraffin were embedded. After the animal experiment was completed, the cell apoptosis was observed by TUNEL uniform staining. Some pieces of ventricular muscle were cut, the size was about 0.5 * 0.5 x 0.5cm for milk. ATP (T1), 30 min (T2), 60 min (T3), 1H (T4) after the end of the flow, 2 h (T5) after the end of the flow (Heart Rate), average arterial pressure, pH value, blood sugar, lactic acid, lactic acid, lactic acid, lactic acid, lactate Lactic Acid, Lac), hemoglobin (Hemoglobin, Hb), samples of fetal sheep venous blood were collected at every time point: CKMB, calcin I (C Tn I) and troponin T (c). All data collected in this study were treated with statistical software. The measurement data were expressed in a mean range of mean standard deviation. The differences between the two groups were significant. By using single factor variance test (Student 's t-test), one-way ANOVA test was used for the significant comparison among multiple groups. Results 1. of the three groups of fetal sheep were in the measured blood gas at every time point, P H, PO_2 (mm Hg), PCO_2 (mm), and afterwards, there was no statistical difference at all time points. HTK group Na+ (mmol/L) is lower than group STH (P0.05), and at T3, HTK group K~+ (mmo/L), Lac (mmol/L), BE (%) is lower than that of the group. 1, P=0.071). The change of serum C Tn I (ng/L) concentration, there is no statistical difference between the three groups when T1, T2, T3, while T4 STH group is higher than before. The number of TUNEL positive cells in group.3.HTK and STH group was higher than that of non stop group (p=0.001 and p=0.048), but the number of TUNEL positive cells in STH group was significantly higher than that in group HTK (p=0.007).4.STH group (p=0.007).4.STH group, and the content of lactic acid in the group was higher than that in the group STH. Compared with STH cardioplegia, HTK solution has obvious advantages in reducing myocardial metabolism and reducing the release of myocardial damage markers in fetal sheep stopping jumping compared with STH cardioplegic solution. TUNEL labeled cell apoptosis and myocardial ATP content have a higher level of ATP content than STH cardioplegia. Increasing the stopping time of.HTK solution is longer than that of STH solution, which can cause hyponatremia.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R654.1

【参考文献】