本文選題：胸段硬膜外阻滯 + 缺血再灌注損傷��； 參考：《新鄉(xiāng)醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的通過對(duì)雄性Wistar大鼠心肌缺血再灌注損傷前給予胸段硬膜外阻滯處理,采集大鼠心率、心電圖及平均動(dòng)脈壓,測(cè)定大鼠各時(shí)間段之間血清心肌肌鈣蛋白I(cTnI)含量,檢測(cè)心肌組織自噬相關(guān)蛋白(Beclin-1)表達(dá)水平及TTC染色計(jì)算心肌梗死面積,來探討胸段硬膜外阻滯對(duì)大鼠心肌缺血再灌注損傷(MIRI)的保護(hù)作用。方法1、研究分組將30只健康成年雄性Wistar大鼠隨機(jī)分為假手術(shù)組、心肌缺血再灌注組(I/R)組和胸段硬膜外阻滯組(TEB)組,每組10只。TEB組、I/R組大鼠結(jié)扎左冠狀動(dòng)脈前降支前分別向硬膜外腔給予體積分?jǐn)?shù)0.5%羅哌卡因0.125ml·kg-1和等量生理鹽水,假手術(shù)組大鼠給予心臟左冠狀動(dòng)脈前降支穿線,但不結(jié)扎,穿線前硬膜外腔給予生理鹽水0.125ml·kg-1;TEB組和I/R組大鼠給予心臟左冠狀動(dòng)脈前降支穿線并結(jié)扎,40min后解開結(jié)扎線復(fù)灌120min。TEB組大鼠胸段硬膜外阻滯后根據(jù)大鼠胸腹肌肉松弛,阻滯區(qū)體表溫度升高,大鼠左前降支結(jié)扎、復(fù)灌后,根據(jù)心肌大體顏色觀察及心電圖等提示動(dòng)物模型制備成功。2、心電圖及血流動(dòng)力學(xué)檢測(cè):利用多功能生物采集系統(tǒng),在大鼠體表連接肢體導(dǎo)聯(lián),持續(xù)檢測(cè)記錄各組大鼠開胸前(T_0)、左冠狀動(dòng)脈結(jié)扎前(T_1)、缺血20min(T_2)、缺血40min(T_3)、再灌注120min(T_4)的心率(HR)、平均動(dòng)脈壓(MAP)。3、ELISA法測(cè)定血清肌鈣蛋白I(cTnI)含量:每只大鼠于開胸前(T_0)、左冠狀動(dòng)脈結(jié)扎前(T_1)、缺血20min(T_2)、缺血40min(T_3)及再灌注120min(T_4)時(shí)取靜脈血,用ELISA法測(cè)血清cTnI水平。4、TTC染色:再灌注120min時(shí)處死大鼠,取心臟,用冰鹽水沖洗干凈積血,將心臟放入-20℃冰箱冷凍20min,切成2mm厚的心肌切片,用TTC染色,用Image—Pro Plus6.0圖象分析軟件(美國(guó)Media Cybernetics公司)計(jì)算梗死區(qū)和心肌缺血危險(xiǎn)區(qū)面積。5、Western blot方法檢測(cè)心肌組織自噬相關(guān)蛋白(Beclin-1)表達(dá)水平:再灌注120min后取心臟,將缺血危險(xiǎn)區(qū)心肌組織放液氮冷凍后,提取組織蛋白,檢測(cè)心肌組織自噬相關(guān)蛋白(Beclin-1)表達(dá)水平。6、統(tǒng)計(jì)分析數(shù)據(jù):采用SPSS21.0統(tǒng)計(jì)軟件進(jìn)行分析,計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差((?)±s)表示,行單因素AVONA方差分析,兩均數(shù)比較采用t檢驗(yàn)P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果1、與組內(nèi)T_0時(shí)及假手術(shù)組同時(shí)間點(diǎn)比較,T_2~T_4時(shí)I/R組、TEB組大鼠HR、MAP顯著降低(P0.05);與I/R組同時(shí)間點(diǎn)比較,T_2~T_4時(shí)TEB組HR、MAP顯著升高(P0.05)。與T_0時(shí)比較T_1時(shí)三組HR、MAP差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。2、與T_0時(shí)比較,T_2~T_4時(shí)I/R組、TEB組血清cTnI水平顯著升高(P0.01);與假手術(shù)組同時(shí)間點(diǎn)比較,T_2~T_4時(shí)I/R、TEB組血清cTnI水平顯著升高(P0.01);與TEB組同時(shí)間點(diǎn)比較,T_2~T_3時(shí)I/R組血清cTnI水平差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),T_4時(shí)I/R組血清cTnI水平較TEB組顯著升高(P0.01),T_0時(shí)三組之間比較血清cTnI水平差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。3、與I/R組比較,TEB組心肌梗死區(qū)與心肌缺血危險(xiǎn)區(qū)的百分比顯著降低(P0.01),假手術(shù)組大鼠無(wú)心肌梗死。4、與假手術(shù)組比較I/R組大鼠心肌組織中Beclin-1表達(dá)顯著增高(P0.01),與I/R組比較TEB組大鼠心肌組織中Beclin-1水平顯著降低(P0.01)。結(jié)論TEB能夠減輕MIRI,對(duì)心肌有明顯的保護(hù)作用,其機(jī)制可能與下調(diào)心肌細(xì)胞自噬因子表達(dá),從而減少再灌注階段心肌細(xì)胞自噬有關(guān)。
[Abstract]:Objective to measure the heart rate, electrocardiogram and mean arterial pressure of rats before myocardial ischemia reperfusion injury in male Wistar rats, and to determine the content of cardiac troponin I (cTnI) in serum, the expression of autophagy related protein (Beclin-1) and TTC staining for myocardial infarction. The protective effect of thoracic epidural block on myocardial ischemia reperfusion injury (MIRI) in rats was investigated. Methods 1, 30 healthy adult male Wistar rats were randomly divided into sham operation group, myocardial ischemia reperfusion group (I/R) group and thoracic epidural block group (TEB) group, 10.TEB groups in each group, and I/R group ligation of left coronary artery. The volume fraction of 0.5% ropivacaine 0.125ml kg-1 and equal amount of normal saline were given to the epidural cavity before the anterior descending branch of the pulse. The rats in the sham operation group were given the left anterior descending branch of the coronary artery, but they were not ligation and 0.125ml. Kg-1 was given to the epidural cavity before the thread was worn, and the left anterior descending branch of the coronary artery in the group of TEB and the group I/R was given the line of the left anterior descending branch of the coronary artery. After ligating and ligation, after 40min, the thoracic and abdominal muscle relaxation of rats in group 120min.TEB was unraveled according to the relaxation of the thoracic and abdominal muscles of the rats, the body surface temperature in the block was increased, the left anterior descending branch of the rat was ligated. After the reperfusion, the animal model was prepared according to the gross color observation of the myocardium and electrocardiogram, and the electrocardiogram and hemodynamic tests were made: using the multifunction test. The biological collection system was used to connect the limb lead on the body surface of the rat. The rats were continuously recorded before the chest (T_0), the left coronary artery ligation (T_1), ischemic 20min (T_2), ischemic 40min (T_3), and reperfusion 120min (T_4) heart rate (HR), mean arterial pressure (MAP).3, and ELISA method to determine the content of serum troponin. Before ligation of the left coronary artery (T_1), ischemic 20min (T_2), ischemic 40min (T_3) and reperfusion 120min (T_4), the venous blood was taken. The serum cTnI level.4, TTC staining was measured by ELISA method. Image - Pro Plus6.0 image analysis software (American Media Cybernetics company) was used to calculate the area.5 in the infarct area and the area of myocardial ischemia. The Western blot method was used to detect the expression level of autophagy related protein (Beclin-1) in myocardial tissue: after reperfusion 120min, the heart was taken, and the tissue protein was extracted after the cryosurgery of the myocardial tissue of the ischemic risk area and detected the tissue protein. Myocardial autophagy related protein (Beclin-1) expression level.6, statistical analysis data: using SPSS21.0 statistical software for analysis, measurement data with mean + standard deviation (?) + s), single factor AVONA variance analysis, two average number comparison using t test P0.05 for difference is statistically significant. Results 1, with the same time in the group T_0 and the sham operation group in the same time Comparison, T_2~T_4 I/R group, group TEB rats HR, MAP significantly decreased (P0.05); compared with the I/R group at the same time point, T_2~T_4 TEB group HR, MAP significantly increased (P0.05). The level of serum cTnI in group TEB was significantly higher in I/R and TEB (P0.01), compared with group TEB at the same time, and there was no significant difference in serum cTnI level in I/R group at T_2~T_3 (P0.05). There was no significant difference in serum levels between the three groups at T_4 (P0.05). Compared with the group TEB, the percentage of myocardial infarction area and myocardial ischemia was significantly lower (P0.01). There was no myocardial infarction in the sham operation group (.4). Compared with the sham group, the Beclin-1 expression in the myocardial tissue of the I/R group was significantly higher (P0.01), and the Beclin-1 level in the myocardial tissue of the TEB group was significantly lower than that of the I/R group (P0.01). Conclusion TEB could be reduced. Light MIRI has an obvious protective effect on myocardium, and its mechanism may be related to downregulation of autophagy factor expression in myocardium, thereby reducing autophagy in myocardial cells at reperfusion stage.

【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R614

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