本文選題：STH液 + HTK液　； 參考：《廣西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的通過觀察比較St.Thomas停搏液(STH液)、組氨酸-色氨酸-酮戊二酸停搏液(HTK液)和不停跳對胎羊?qū)嵭畜w外循環(huán)(CPB)術(shù)中及術(shù)后心肌酶、心肌組織結(jié)構(gòu)、代謝及實驗中其它相關(guān)指標(biāo)的影響,評估這三種方法對胎羊心肌的保護作用,尋求一種適合的胎羊體外循環(huán)心肌的保護方法,從而為臨床胎兒體外循環(huán)提供依據(jù)。方法18只懷孕山羊(孕100—120 d,足月為150 d),重量為25.72±4.29 kg(17.5-32 kg),共孕有18只胎羊。分為3組:不停跳組(不停跳組),STH組(St.Thomas保護液組),HTK組(HTK保護液組)。3組胎羊均采用胸骨前正中切口,完全肝素化后建立CPB行主肺動脈-右心房插管轉(zhuǎn)流,體外循環(huán)回路建立好后開始體外循環(huán),其中St.Thomas保護液組和HTK保護液組在體外循環(huán)開始后,用主動脈鉗夾閉主動脈開始灌流心肌保護液,不停跳組不灌流心肌保護液�？刂铺パ蝮w外循環(huán)流量在200 ml/kg/min以上,水箱溫度恒定40℃。St.Thomas保護液組和HTK保護液組使胎羊心臟停跳30分鐘,然后開放升主動脈在并行循環(huán)下使胎羊心臟復(fù)跳,30分鐘后撤離體外循環(huán),觀察2小時,不停跳組轉(zhuǎn)流60 min后停機觀察2h。在CPB前、轉(zhuǎn)流30min、轉(zhuǎn)流60min、停機后1h、停機后2h采頸靜脈血5ml,腋動脈動脈血1ml(測血氣)。胎羊靜脈血均在4℃下4000 rpm離心15min,取上清液,分裝入EP管(每管約0.5ml),放入-80℃冰箱凍存。在停機后2h,胎羊靜脈注射10%氯化鉀5 ml,使心跳迅速停跳,剪取胎羊右心房心肌組織大小約1×1×1cm,置入含有4%福爾馬林液的試管中密封保存于常溫中,待1周后進行脫水、石蠟包埋,動物實驗完成后用TUNEL統(tǒng)一染色觀察細(xì)胞凋亡情況,剪取心室肌若干塊,大小約0.5×0.5×0.5cm用于乳酸和ATP的檢測。分別于轉(zhuǎn)流前(T1)、轉(zhuǎn)流30 min(T2)、轉(zhuǎn)流60 min(T3)、轉(zhuǎn)流結(jié)束后1h(T4)、轉(zhuǎn)流結(jié)束后2 h(T5)測心率(Heart Rate,HR)、平均動脈壓(Mean Artery Pressure,MAP)、體外循環(huán)轉(zhuǎn)流量、PH值、PaO_2、Pa CO_2、堿剩余(Base Excess,BE)、K~+、Na~+、Ca~(2+)、血糖(Glucose,Glu)、乳酸(Lactic Acid,Lac)、血紅蛋白(Hemoglobin,Hb),在各個時間點采集胎羊靜脈血檢驗心肌受損指標(biāo):CKMB、鈣蛋白I(c Tn I)和肌鈣蛋白T(c Tn T)。本次研究采集的所有數(shù)據(jù)均用SPSS21.0統(tǒng)計學(xué)軟件處理,計量資料以均數(shù)±標(biāo)準(zhǔn)差(x±s)的方法表示。兩組間差異顯著性檢驗利用單因素方差檢驗(Student’s t-test),多組間顯著性比較采用one-way ANOVA檢驗。結(jié)果1.三組胎羊在各個時間點腋動脈在測得的血氣中,p H、PO_2(mm Hg),PCO_2(mm Hg)、K~+(mmol/L)、Ca2+(mmol/L)在各時間點比較均無統(tǒng)計學(xué)差異(P0.05)。在T3及T3以后的時段HTK組Na+(mmol/L)要低于STH組(P0.05);在T3時,HTK組的K~+(mmo/L)、Lac(mmol/L)、BE(mmol/L)濃度低于STH組(P0.05);T4時,HTK組的Hct(%)高于STH組,有統(tǒng)計學(xué)差異(P0.05)。2.體外循環(huán)之前胎羊血清c Tn T、c Tn I、CKMB濃度分別在三組間比較均無明顯差異(P=0.430,P=0.391,P=0.071)。血清c Tn I(ng/L)濃度變化,在T1、T2、T3時三組間比較無統(tǒng)計學(xué)差異(P0.05);T4時STH組的c Tn I較CPB前明顯升高(P0.05);T5時不停跳組與HTK組的c Tn I均較比CPB前升高(P0.05);T4和T5時HTK組的c Tn I低于STH組(P0.05);T4時STH組和T5時的STH組、HTK組的c Tn I高于不停跳組(P0.05)。3.HTK組和STH組TUNEL陽性細(xì)胞數(shù)均較不停跳組組明顯增多(p=0.001和p=0.048),但STH組TUNEL陽性細(xì)胞數(shù)較HTK組顯著增多(p=0.007)。4.STH組心肌組織ATP含量較不停跳組與HTK降低(P0.05);HTK組的乳酸含量高于不停跳組(P0.05);STH組的乳酸含量高于不停跳組和HTK組(P0.05)結(jié)論胎羊體外循環(huán)中,不停跳對心肌的保護明顯優(yōu)于停跳。在胎羊停跳方面與STH停搏液相比,HTK液在降低心肌代謝、減少心肌損傷標(biāo)記物的釋放方面有明顯優(yōu)勢,TUNEL標(biāo)記的細(xì)胞凋亡減少,心肌ATP含量有增加。HTK液的停跳時間要長與STH液相比可引起低鈉血癥。
[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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R654.1

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