本文選題：異氟醚 + 血糖　； 參考：《南京大學(xué)》2017年碩士論文

【摘要】：研究背景:圍術(shù)期高血糖可引起嚴(yán)重的術(shù)后預(yù)后不良,增加院內(nèi)并發(fā)癥如感染、急性腎損傷等的發(fā)病率以及術(shù)后30天和1年死亡率。通常認(rèn)為圍術(shù)期高血糖的發(fā)生與患者術(shù)前基礎(chǔ)疾病、術(shù)中手術(shù)操作、術(shù)前與術(shù)中用藥等相關(guān),越來越多的基礎(chǔ)及臨床研究表明麻醉藥也參與了圍術(shù)期高血糖的發(fā)生,吸入麻醉藥七氟醚和異氟醚可引起機(jī)體葡萄糖利用受損、糖耐量下降、胰島素分泌減少、胰島素抵抗。下丘腦是機(jī)體能量調(diào)節(jié)中心,參與了體內(nèi)多種物質(zhì)如糖、脂的代謝,腺苷酸活化蛋白激酶(adenosine monophosphate activated protein kinase,AMPK)在其調(diào)節(jié)過程中起主要作用,目前關(guān)于下丘腦在吸入麻醉藥與血糖的代謝中的作用尚未有研究。二甲雙胍是目前一線降糖藥,有著良好的降糖效果和安全性,其降糖作用也與AMPK相關(guān)。本研究通過對(duì)健康小鼠行異氟醚麻醉觀察小鼠血糖的變化,使用二甲雙胍來研究圍術(shù)期高血糖的預(yù)防,檢測(cè)下丘腦AMPK mRNA的表達(dá)探索相關(guān)機(jī)制。研究方法:本研究使用的動(dòng)物類型為C57BL/6的小鼠、雄性、8周齡,體重約22-28 g。采用持續(xù)吸入6 h異氟醚麻醉,通過尾靜脈取血加血糖儀來檢測(cè)血糖,使用生理鹽水溶解二甲雙胍后通過腹腔給藥。血糖檢測(cè)時(shí)間點(diǎn)分別為麻醉前0.5 h,麻醉0.5h、1h、2h、3h、4h、6h,麻醉結(jié)束后2h、4h。本研究實(shí)驗(yàn)過程主要分為以下三部分:實(shí)驗(yàn)一研究不同濃度異氟醚麻醉對(duì)小鼠血糖的影響將隨機(jī)分為未禁食組(non-fasted group)和禁食12 h組(fasted group),各組再隨機(jī)分為兩組:對(duì)照組(Cn組/Cf組),麻醉組(An組/Af組)。使用濃度為1.2%或1.8%異氟醚分別進(jìn)行麻醉,檢測(cè)麻醉下小鼠血糖水平。實(shí)驗(yàn)二研究不同劑量二甲雙胍對(duì)異氟醚麻醉下小鼠血糖的影響根據(jù)實(shí)驗(yàn)一結(jié)果,選擇與對(duì)照組血糖值差異較大的濃度進(jìn)行下一步實(shí)驗(yàn)。同樣設(shè)置禁食組和未禁食組,各組再隨機(jī)分為四組:對(duì)照組(Cn組/Cf組),麻醉組(An組/Af組),二甲雙胍組(Mn組/Mf組)和二甲雙胍+麻醉組(MAn組/MAf組)。二甲雙胍組和二甲雙胍+麻醉組于麻醉前1 h腹腔注射二甲雙胍40 mg/kg,麻醉組和二甲雙胍+麻醉組均行異氟醚6 h麻醉。二甲雙胍組(Mn組/Mf組)和二甲雙胍+麻醉組(MAn組/MAf組)。二甲雙胍組和二甲雙胍+麻醉組于麻醉前1 h腹腔注射二甲雙胍20 mg/kg或40 mg/kg,麻醉組和二甲雙胍+麻醉組均行異氟醚6 h麻醉。通過尾靜脈取血檢測(cè)圍麻醉期各時(shí)間點(diǎn)血糖值。實(shí)驗(yàn)三研究AMPKα1和AMPKα2在二甲雙胍預(yù)處理對(duì)異氟醚麻醉后小鼠血糖變化中的機(jī)制關(guān)聯(lián)根據(jù)實(shí)驗(yàn)二結(jié)果,選擇對(duì)血糖影響較大的二甲雙胍給藥劑量組設(shè)立平行取材組,每組4只,于麻醉1 h、4 h、6 h取材下丘腦,通過Q-PCR檢測(cè)AMPKα1、AMPKα2 mRNA的表達(dá)水平。研究結(jié)果:實(shí)驗(yàn)一:1)基礎(chǔ)值比較:本實(shí)驗(yàn)結(jié)果表明禁食12h后小鼠基礎(chǔ)血糖值為6.5±0.7mmol/L,未禁食組為8.3±1.0 mmol/L,二者比較有顯著差異(P0.001)。2)麻醉對(duì)血糖的影響:異氟醚1.0 MAC(1.2%)組:與Cn組相比,1.2%異氟醚麻醉可在麻醉0.5 h-4 h引起An組小鼠血糖升高;而與Cf組相比,Af組小鼠血糖在麻醉0.5 h,3 h、4 h顯著升高(P0.05)。異氟醚1.5 MAC(1.8%)組:與Cn組相比,An組小鼠血糖在麻醉0.5 h、2 h顯著升高(P0.05);與Cf組相比,Af組未出現(xiàn)升高,在麻醉4h和6h出現(xiàn)降低(P0.05)。實(shí)驗(yàn)二:結(jié)果一表明,異氟醚濃度為1.0 MAC時(shí)小鼠血糖變化更大,因此本此實(shí)驗(yàn)選擇1.2%異氟醚濃度進(jìn)一步探索二甲雙胍對(duì)異氟醚麻醉下小鼠血糖的影響。本次研究中單純異氟醚麻醉組血糖變化與結(jié)果一一致,單獨(dú)給予不同劑量二甲雙胍組小鼠與各自對(duì)照組無明顯差異(P0.05)。1)預(yù)給藥二甲雙胍為20 mg/kg的分組:與Cn組相比,An組小鼠在麻醉3 h出現(xiàn)顯著升高的血糖水平(P0.05),隨后恢復(fù)至與Cn組相同的水平(P0.05),而MAn組小鼠血糖在麻醉1-2 h顯著升高(P0.05),隨后逐漸降低至與Cn組相同的水平;與Cf組相比,Af組小鼠在麻醉0.5-2 h出現(xiàn)顯著升高的血糖水平,隨后在麻醉結(jié)束至麻醉結(jié)束2h顯著低于Cf組(P0.05),于麻醉結(jié)束3 h后回到與Cf組相同的水平(P0.05);MAf組小鼠在麻醉0.5-1 h出現(xiàn)顯著升高的血糖水平,隨后在麻醉結(jié)束至麻醉結(jié)束2 h顯著低于Cf組(P0.05),于麻醉結(jié)束3 h后回到與Cf組相同的水平(P0.05)。2)預(yù)給二甲雙胍為40 mg/kg的分組:與Cn組和Cf組相比,An組和Af組分別在麻醉0.5h、1h、2h和1h,2h,3h升高(P0.05);隨后出現(xiàn)血糖水平下降,An組在麻醉結(jié)束至結(jié)束后3 h期間血糖水平低于Cn組(P0.05),Af組血糖降低,在麻醉4 h后與Cf組無統(tǒng)計(jì)學(xué)差異(P0.05)。與對(duì)照組相比,單純二甲雙胍處理Mn組和Mf組血糖水平無明顯差異(P0.05),合并異氟醚麻醉的MAn組和MAf組均于麻醉2 h開始出現(xiàn)低血糖并持續(xù)(P0.05),這種差異分別在麻醉結(jié)束4h和1h后消失。實(shí)驗(yàn)三:根據(jù)實(shí)驗(yàn)二結(jié)果表明,40 mg/kg二甲雙胍對(duì)麻醉小鼠血糖影響更為顯著,因此,本次實(shí)驗(yàn)選擇劑量為40 mg/kg二甲雙胍設(shè)立平行取材組。下丘腦AMPK mRNA表達(dá):1)麻醉1 h取材組:與Cf組相比,MAf組AMPKα1和AMPKα 2 mRNA表達(dá)下調(diào)(P0.05)。2)麻醉4 h取材組:與Cn組和Cf組比較,An組、MAn組和Af組AMPKα1表達(dá)下調(diào)(P0.05),AMPKα2mRNA表達(dá)在MAf組降低(P0.05)。3)麻醉6 h取材組:與Cn組和Cf組比較,AMPKα1和AMPKα2在An組和MAn組,Af組和MAf組mRNA表達(dá)都降低(P0.05)。實(shí)驗(yàn)結(jié)論:長時(shí)程異氟醚麻醉可使小鼠血糖呈"先高后低"改變,二甲雙胍預(yù)處理后麻醉小鼠可出現(xiàn)持續(xù)低血糖,這一過程與下丘腦AMPK轉(zhuǎn)錄活性無明顯關(guān)聯(lián)。
[Abstract]:Background: perioperative hyperglycemia may cause severe postoperative poor prognosis, increase the incidence of intraoperative complications such as infection, acute renal injury, and the mortality rate of 30 and 1 years after operation. Basic and clinical studies have shown that anesthetics also participate in the occurrence of hyperglycemia in the perioperative period. Inhalation of anesthetic sevoflurane and isoflurane can cause impaired glucose utilization, decrease of glucose tolerance, decrease of insulin secretion, insulin resistance. The hypothalamus is the body's energy regulation center, and participates in various substances in the body such as sugar, lipid metabolism, adenylate activation. Adenosine monophosphate activated protein kinase (AMPK) plays a major role in its regulation, and the role of the hypothalamus in the metabolism of inhaled anesthetics and blood glucose has not been studied. Metformin is a frontline hypoglycemic agent at present. It has a good effect on reducing sugar and safety. The hypoglycemic effect of metformin is also associated with AMPK. In this study, the changes of blood glucose in mice were observed by isoflurane anesthesia in healthy mice. Metformin was used to study the prevention of hyperglycemia in the perioperative period and to detect the mechanism of the expression of AMPK mRNA in the hypothalamus. The research method: This study was used in mice of C57BL/6, male, 8 weeks of age, and weight about 22-28 g. by continuous inhalation of 6 h The blood glucose was detected by the caudal vein and the blood glucose meter through the caudal vein. The time point of blood glucose detection was 0.5 h before anesthesia, 0.5h, 1H, 2h, 3h, 4h, 6h, 2h in the end of anesthesia, 4h. in the following three parts: the experiment studied the different concentrations of isofluorine in the experiment. The effects of ether anesthesia on blood glucose in mice were randomly divided into unfasting group (non-fasted group) and fasting 12 h group (fasted group). Each group was then randomly divided into two groups: control group (group Cn /Cf) and anesthetic group (group /Af group An). The blood glucose level of mice under anesthesia was detected with concentration of 1.2% or 1.8% isoflurane respectively. The experiment two studied different doses. The effect of metformin on the blood sugar of mice under isoflurane anesthesia was selected according to the results of the experiment, and the concentration of blood sugar in the control group was selected for the next step. The same group was set up in four groups: control group (group /Cf of Cn group), anesthesia group (Group An /Af), metformin group (group /Mf of Mn group) and two a pair Guanidine + anaesthesia group (group MAn /MAf). The metformin group and metformin + anaesthesia group were intraperitoneally injected with metformin 40 mg/kg 1 h before anesthesia. The anesthesia group and metformin + anaesthesia group were treated with isoflurane 6 h anesthesia. Metformin group (group Mn /Mf group) and metformin + anaesthesia group (group /MAf group MAn). Metformin group and metformin + anaesthesia group were 1 before anesthesia. H intraperitoneal injection of metformin 20 mg/kg or 40 mg/kg, anesthesia group and metformin + anaesthesia group were treated with isoflurane 6 h anesthesia. Blood glucose values were measured at various time points in the perioperative period through the tail vein extraction. Experiment three study the mechanism of AMPK alpha 1 and AMPK alpha 2 on the changes of blood sugar in mice after isoflurane anaesthetized by metformin pretreatment, according to experimental two results The group of metformin, which had more influence on blood glucose, set up a parallel group, 4 rats in each group, 1 h, 4 h, 6 h in the hypothalamus, and the expression level of AMPK alpha 1, AMPK alpha 2 mRNA by Q-PCR. The result of the study: experiment one: 1) base value comparison: the results of this experiment showed that the base blood sugar of the mice after 12h was 6.5 + 0.7mmol/L, no prohibition. The food group was 8.3 + 1 mmol/L, and the two were significantly different (P0.001).2) the effect of anaesthesia on blood sugar: Isoflurane 1 MAC (1.2%) group: compared with the Cn group, 1.2% isoflurane anesthesia could increase the blood sugar of the An mice in 0.5 H-4 h; compared with the Cf group, the blood sugar in the Af group was significantly increased in 0.5 h, 3 h, and 4. (1.8%). Group: compared with group Cn, blood glucose of mice in group An was 0.5 h and 2 h increased significantly (P0.05). Compared with group Cf, there was no increase in Af group and decreased in 4H and 6h in anaesthesia (P0.05). Experiment two: the results showed that the blood glucose in mice was higher when the concentration of isoflurane was 1 MAC, because this experiment selected 1.2% isoflurane concentration to further explore metformin. In this study, the blood sugar changes in the isoflurane anaesthetized group were consistent with the results. There was no significant difference between the different doses of metformin group and the control group (P0.05).1). The group of pre administered metformin was 20 mg/kg: compared with the Cn group, the mice in the An group increased significantly in the 3 h anesthesia. The blood glucose level (P0.05) was then restored to the same level as that in the Cn group (P0.05), while the blood glucose in the MAn group increased significantly (P0.05) at 1-2 h (P0.05), and then gradually decreased to the same level as that in the Cn group. Compared with the Cf group, the serum sugar level of the Af group mice was significantly higher in the 0.5-2 h, and the 2H was significantly lower than that of the group of 0.5-2. 0.05) back to the same level as that in the Cf group after the end of the anesthesia (P0.05); in group MAf mice, the level of blood glucose increased significantly in the anesthetic 0.5-1 h, and then at the end of anesthesia to the end of anesthesia 2 h was significantly lower than that of the Cf group (P0.05), and returned to the same level as the Cf group after the end of the anesthesia 3 h (P0.05). Compared with group Cf, group An and Af group were anesthetized 0.5h, 1H, 2h and 1H, 2h, 3h increased (P0.05), and then decreased blood sugar levels. The glucose level in the An group was lower than that in the 3 h group after the end of anesthesia. There was no significant difference in blood glucose level in group Mf (P0.05). The MAn group and MAf group with isoflurane anesthesia began to appear hypoglycemia and continued (P0.05) at 2 h anesthesia. This difference disappeared after the end of anesthesia 4H and 1H. Experiment three: according to experimental two, 40 mg/kg two metformin had a more significant effect on the blood glucose in the anesthetized mice. Therefore, this experiment 40 mg/kg metformin was selected to set up a parallel selection group. The expression of AMPK mRNA in the hypothalamus: 1) anaesthetized 1 h group: compared with the Cf group, the AMPK alpha 1 and AMPK a 2 mRNA expression of 2 mRNA (P0.05).2) were anesthetized 4 h materials. 6 h anesthesia group: compared with group Cn and Cf group, AMPK alpha 1 and AMPK alpha 2 in group An and MAn group, Af and MAf group mRNA expression decreased (P0.05). Experimental conclusion: long term isoflurane anesthesia can make the blood glucose in mice "first high and low" change, metformin pretreated mice can appear continuous hypoglycemia, this process and hypothalamic AMPK transcriptional activity There is no obvious connection.

【學(xué)位授予單位】：南京大學(xué)
【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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本文編號(hào)：1810678