本文選題：亞低溫 + 心臟驟停��； 參考：《蘇州大學(xué)》2016年博士論文

【摘要】：第一部分:大鼠心肺復(fù)蘇后亞低溫模型的建立與評(píng)估目的:利用窒息法使大鼠心臟驟停后行心肺復(fù)蘇,在復(fù)蘇后使用冰袋誘導(dǎo)低溫,并評(píng)估大鼠心肺復(fù)蘇后的亞低溫模型。方法:將10%水合氯醛(0.3 ml/100 g)注射進(jìn)腹腔內(nèi)用于麻醉,然后氣管切開放置氣管插管,將套管插到股動(dòng)脈和股靜脈。然后檢測動(dòng)脈血壓以及記錄標(biāo)準(zhǔn)Ⅱ號(hào)導(dǎo)聯(lián)動(dòng)態(tài)心電圖。在距離肛門1.5厘米處放置探針并使用電子溫度計(jì)來檢測直腸溫度。手術(shù)后,讓大鼠情緒穩(wěn)定15分鐘,然后去除心率和血壓明顯異常的大鼠。夾住氣管,使大鼠突然心臟驟停(心電圖表現(xiàn)為心室顫動(dòng),無脈性電活動(dòng)或心搏停止,或收縮壓下降到25 mm Hg以下并且動(dòng)脈搏動(dòng)消失)。6分鐘后,將夾住的氣管松開,并且連接動(dòng)物呼吸機(jī)為其輔助通氣,同時(shí)使用胸外心臟按壓,如果有心室纖維性顫動(dòng),使用電除顫和利多卡因靜脈注射。當(dāng)自主循環(huán)恢復(fù)(自主循環(huán)標(biāo)準(zhǔn)為HR?170次/分且收縮壓?90 mm Hg),并且自主循環(huán)維持5分鐘后,終止按壓。如果在心肺復(fù)蘇6分鐘后,自主循環(huán)沒有恢復(fù),復(fù)蘇終止。當(dāng)自主呼吸恢復(fù)時(shí),撤除呼吸機(jī)。在監(jiān)測到自主循環(huán)恢復(fù)35分鐘后,移除股動(dòng)脈以及靜脈插管,縫合。當(dāng)大鼠蘇醒時(shí),立即0.12 ml/100 g濃度為10%的水合氯醛注入腹腔內(nèi)。在復(fù)蘇后,對(duì)照組在室溫下(25℃)治療,而亞低溫兩組使用冰袋誘導(dǎo)低溫,并且在誘導(dǎo)體溫降低后的30分鐘內(nèi),達(dá)到最佳溫度(33/30℃)。如果體溫偏離最佳溫度,予以調(diào)節(jié)。如果體溫高于預(yù)期標(biāo)準(zhǔn),噴稀乙醇或者再結(jié)合電風(fēng)扇使其降低溫度。當(dāng)大鼠的體溫低于預(yù)期標(biāo)準(zhǔn),將大鼠放于室溫的環(huán)境下使其回溫。體溫的波動(dòng)幅度要控制在±0.3℃內(nèi)。結(jié)果:大鼠在心肺復(fù)蘇后的30分鐘內(nèi),亞低溫組分別達(dá)到輕度及中度低溫溫度(33/30℃)。結(jié)論:大鼠心肺復(fù)蘇后輕度及中度低溫模型建立成功。第二部分:亞低溫治療對(duì)心肺復(fù)蘇后大鼠腦組織凋亡相關(guān)蛋白的作用目的:探討輕度和中度低溫治療對(duì)心肺復(fù)蘇后大鼠腦組織凋亡相關(guān)蛋白的作用。方法:30只SD大鼠被隨機(jī)分成對(duì)照組(正常體溫),體溫為33℃輕度低溫組以及體溫為30℃中度低溫組,每組各10只。對(duì)照組大鼠在進(jìn)行心肺復(fù)蘇后,25℃室溫下接受常規(guī)治療,而輕度低溫組以及中度低溫組的大鼠經(jīng)過心肺復(fù)蘇后,分別接受33℃和30℃低溫治療。所有組經(jīng)過心肺復(fù)蘇24小時(shí)后,將腦組織取出進(jìn)行研究,并利用免疫組化法檢測大腦皮層的凋亡相關(guān)蛋白caspase-3,用蛋白免疫印跡法(western blot)檢測凋亡相關(guān)蛋白Bcl-2以及Bax的表達(dá)。結(jié)果:與對(duì)照組相比,大腦神經(jīng)元亞低溫組的caspase-3明顯下降(p0.01),體溫為30℃的低溫組與33℃低溫組相比caspase-3明顯更低,但無統(tǒng)計(jì)學(xué)意義(p?0.05);Bcl-2的表達(dá)在亞低溫組有明顯的增加(p0.01),并且體溫為30℃低溫組的增加程度高于體溫為33℃的低溫組(p0.05);而Bax的表達(dá)在這三組中無明顯的區(qū)別。結(jié)論:輕度及中度低溫可以下調(diào)caspase-3,上調(diào)Bcl-2的表達(dá)。顯示了低溫在心肺復(fù)蘇之后有保護(hù)大腦的作用。并且30℃的低體溫療法比33℃的低溫療法有更好的治療作用。第三部分:亞低溫治療對(duì)心肺復(fù)蘇后大鼠腦組織膠質(zhì)纖維酸性蛋白(GFAP)的作用目的:探討亞低溫治療對(duì)心肺復(fù)蘇后大鼠腦組織膠質(zhì)纖維酸性蛋白(GFAP)的作用。方法:30只SD大鼠隨機(jī)分成對(duì)照組(正常體溫),體溫為33℃的低溫組以及體溫為30℃低溫組,每組各10只。對(duì)照組的大鼠在進(jìn)行心肺復(fù)蘇后,在25℃室溫下接受常規(guī)治療,而輕度低溫組以及中度低溫組的大鼠,經(jīng)過心肺復(fù)蘇后在亞低溫下進(jìn)行治療。在所有組經(jīng)過心肺復(fù)蘇24小時(shí)后,將腦組織取出研究,并利用免疫組化法檢測大腦皮層的膠質(zhì)纖維酸性蛋白(GFAP)表達(dá)。結(jié)果:與對(duì)照組相比,膠質(zhì)纖維酸性蛋白的表達(dá)在亞低溫組中有明顯的增加(p0.01)。結(jié)論:輕度及中度低溫通過減少caspase-3的表達(dá)以及增加Bcl-2的表達(dá)去調(diào)控神經(jīng)膠質(zhì)纖維酸性蛋白的表達(dá),引起膠質(zhì)纖維酸性蛋白的表達(dá)上調(diào),從而促進(jìn)大腦細(xì)胞信號(hào)轉(zhuǎn)導(dǎo),進(jìn)一步抑制細(xì)胞凋亡,減少腦損傷。
[Abstract]:The first part: the establishment and evaluation of hypothermia model in rats after cardiopulmonary resuscitation: using asphyxia method to make cardiopulmonary resuscitation after cardiac arrest, using ice bag to induce hypothermia after resuscitation, and evaluate the hypothermia model after cardiopulmonary resuscitation in rats. Method: injection of 10% chloral chloral (0.3 ml/100 g) into the abdominal cavity for anesthesia, and then trachea The cannula was inserted into the endotracheal intubation and inserted into the femoral and femoral veins. Then the arterial blood pressure and the standard II lead dynamic electrocardiogram were detected. A probe was placed at 1.5 cm from the anus and an electronic thermometer was used to detect the rectal temperature. After surgery, the rats were emotionally stabilized for 15 minutes, and then the heart rate and blood pressure were significantly abnormally removed. Rats were trapped in the trachea, causing sudden cardiac arrest in rats (the electrocardiogram showed ventricular fibrillation, no pulse electrical activity or cardiac arrest, or the systolic pressure dropped below 25 mm Hg and the arterial pulsation disappeared). After.6 minutes, the trapped trachea was loosened and the animal ventilator was connected to its auxiliary ventilation, and the chest compression was used, if the heart was pressed, if the heart was in mind. Ventricular fibrillation, using electric defibrillation and lidocaine intravenous injection. When autonomic circulation recovery (independent circulation standard is HR? 170 / sub and systolic pressure? 90 mm Hg), and the autonomic circulation is maintained for 5 minutes, the press is terminated. If the cardiopulmonary resuscitation is 6 minutes, the autonomic circulation is not recovered and the resuscitation terminates. When the self respiration recovery is recovered, the ventilator is removed. After 35 minutes of recovery, the femoral artery and venous cannula were removed and sutured. When the rat was awakened, the 0.12 ml/100 G concentration of chloral hydrate was injected into the abdominal cavity immediately. After resuscitation, the control group was treated at room temperature (25 degrees C), while the hypothermia two groups used ice bags to induce hypothermia and 30 minutes after the hypothermia was induced. Inside, reach the best temperature (33/30 C). If the temperature deviates from the best temperature, adjust it. If the body temperature is higher than the expected standard, spray dilute ethanol or combine the electric fan to reduce the temperature. When the body temperature of the rat is lower than the expected standard, the rat is put back at room temperature. The fluctuation range of body temperature is controlled within 0.3. In 30 minutes after cardiopulmonary resuscitation, mild to moderate hypothermia temperatures (33/30 C) were reached in the hypothermia group. Conclusion: the mild to moderate hypothermia model after cardiopulmonary resuscitation in rats was established successfully. The second part: the effect of mild hypothermia treatment on apoptosis related egg white in the brain tissue of rats after cardiopulmonary resuscitation: the treatment of mild and moderate hypothermia treatment Methods: 30 SD rats were randomly divided into control group (normal body temperature), mild hypothermia at 33 centigrade and 10 moderate hypothermia group at 30 centigrade, 10 rats in each group. The control group received routine treatment at the temperature of 25, and mild hypothermia after cardiopulmonary resuscitation, and the mild hypothermia group. After cardiopulmonary resuscitation, rats in the moderate hypothermia group received 33 and 30 centigrade cryogenic treatment. After 24 hours of cardiopulmonary resuscitation, the brain tissue was taken out and the apoptosis related protein caspase-3 in the cerebral cortex was detected by immunohistochemistry. The apoptosis related protein Bcl-2 was detected by protein immuno trace (Western blot) method. Results: compared with the control group, compared with the control group, the caspase-3 of the hypothermia group of the cerebral neurons decreased significantly (P0.01), the low temperature group at 30 C was significantly lower than the 33 C group, but there was no statistical significance (P? 0.05); the expression of Bcl-2 was significantly increased in the sub hypothermia group (P0.01), and the body temperature was 30 centigrade cryogenic group. The degree is higher than the hypothermia group (P0.05) at 33 degrees centigrade; and the expression of Bax is not distinctly different in these three groups. Conclusion: mild and moderate hypothermia can down regulate the expression of Caspase-3 and increase the expression of Bcl-2. It shows that the hypothermia can protect the brain after cardiopulmonary resuscitation, and the low body temperature therapy at 30 C has better treatment than the cryotherapy at 33. The third part: the effect of mild hypothermia on glial fibrillary acidic protein (GFAP) in the brain tissue of rats after cardiopulmonary resuscitation: To explore the effect of hypothermia therapy on glial fibrillary acidic protein (GFAP) in the brain tissue of rats after cardiopulmonary resuscitation. Methods: 30 SD rats were randomly divided into two groups (normal body temperature), hypothermia group at 33 C and body temperature. After cardiopulmonary resuscitation, the rats in the control group received routine treatment at the room temperature of 25, while the rats in the control group received routine treatment at 25 C, while the mild hypothermia group and the moderate hypothermia group were treated at subhypothermia after cardiopulmonary resuscitation. After 24 hours of cardiopulmonary resuscitation, the brain tissue was removed and immuno histochemical was used in all groups. Results: the expression of glial fibrillary acidic protein (GFAP) in the cerebral cortex was detected. Results: compared with the control group, the expression of glial fibrillary acidic protein was significantly increased in the hypothermia group (P0.01). Conclusion: mild to moderate hypothermia can regulate the expression of glial fibrillary acidic protein by reducing the expression of Caspase-3 and increasing the expression of Bcl-2. Glial fibrillary acidic protein expression is upregulated, thereby promoting signal transduction in brain cells, further inhibiting cell apoptosis and reducing brain damage.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R614

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