本文選題：姜黃素 + 呼吸　； 參考：《河北醫(yī)科大學》2014年碩士論文

【摘要】：目的：單肺通氣（OLV）是指患者開胸后只利用非手術側肺進行通氣的方式。OLV時非通氣側肺萎陷，為術者提供良好視野方便操作的同時還具有防止兩肺間交叉感染等優(yōu)點。但單肺通氣時間過長可以誘發(fā)急性肺損傷[1]。姜黃素是自姜黃根莖中提取的活性成分，研究表明姜黃素可減輕單肺通氣所誘發(fā)的肺損傷[2]，但作用機制尚不清楚。核因子NF-E2相關因子2（Nrf2）是機體內調節(jié)氧化/還原反應和炎性反應的關鍵性轉錄因子，研究發(fā)現(xiàn)Nrf2在急性肺損傷中發(fā)揮著重要的作用[3-4]。本研究擬評價姜黃素預先給藥對單肺通氣兔肺組織Nrf2蛋白表達的影響，為明確姜黃素減輕單肺通氣所誘發(fā)肺損傷機制提供參考。 方法：健康成年雄性新西蘭大白兔24只，體重2.5~3.0kg，由河北醫(yī)科大學實驗動物中心提供。采用隨機數(shù)字表法，將其隨機分為3組（n=8）：雙肺通氣組（TLV組）、右肺單肺通氣組（OLV組）、姜黃素預先給藥組（Cur組）。 取兔稱重后經(jīng)右耳緣靜脈注射3%戊巴比妥鈉30mg/kg麻醉，麻醉成功后游離右股動脈行穿刺置管術用于監(jiān)測平均動脈壓（MAP）及術中采集動脈血樣。2%鹽酸利多卡因局部浸潤麻醉下切開頸部皮膚，游離右頸內靜脈行穿刺置管術用于術中補液及給藥。選擇ID3.0mm氣管導管，行氣管切開術后，TLV組氣管導管置入氣管行雙肺通氣，OLV組和Cur組氣管導管置入右主支氣管行單肺通氣。通過觀察右側胸廓運動的變化、聽診雙肺呼吸音及監(jiān)測氣道壓力的變化來判斷和調整導管的位置。氣管插管成功后，TLV組雙肺通氣3h，OLV組和Cur組右肺單肺通氣3h，3組動物均采用容量通氣模式，F(xiàn)iO2100%，調整通氣參數(shù)維持SpO2＞90%。所有動物均右側臥位，穩(wěn)定10min后右頸內靜脈注射維庫溴銨0.1mg/kg，待自主呼吸消失后連接呼吸機行機械通氣。OLV組和Cur組均先單肺通氣50min，再雙肺通氣10min（通氣參數(shù)不變，將導管退至氣管內），再重復上述通氣過程。同時兩組均在左側胸壁5~6肋間處開一大小約1cm小口至胸腔，用于觀察左肺組織的萎陷及復張情況。實驗過程中間斷靜脈給予維庫溴銨0.1mg/kg維持肌松，必要時靜脈追加3%戊巴比妥鈉1ml維持麻醉，以10ml·kg-1·h-1的速率自右頸內靜脈持續(xù)泵注乳酸鈉林格氏液，以補充實驗過程中所造成的體液丟失，維持MAP穩(wěn)定。于通氣開始即刻（T0）、通氣1、2、3h（T1-3）采集右股動脈動脈血樣進行血氣分析，測定PaO2并計算氧合指數(shù)（OI），計算公式：OI=PaO2/FiO2。通氣結束后采用股動脈快速放血法處死動物，取雙肺下葉約1cm×1cm×2cm大小肺組織并一分為二，一半用液氮凍存24h后轉80oC冰箱保存待測，另一半用4oC4%多聚甲醛溶液固定待測。將剩余肺組織稱濕重后置于烤箱中（80℃，72h）烘烤，恒重后稱干重，計算雙肺濕/干重比（W/D比）。 取4%多聚甲醛溶液中固定肺組織，常規(guī)脫水、石蠟包埋、切片及HE染色，光學顯微鏡下觀察病理學結果并行肺組織病理學損傷評分，免疫組織化學法檢測兔肺組織Nrf2蛋白表達。80oC保存肺組織采用化學比色法檢測丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性及Western blot法檢測兔肺組織Nrf2蛋白表達。 結果：1與TLV組比較，OLV組及Cur組T2,3時OI下降（P＜0.05）；與T0時比較，OLV組及Cur組T2,3時OI下降（P＜0.05）；與OLV組比較，Cur組T3時OI升高（P＜0.05）。 2光鏡下TLV組雙側肺組織肺泡結構基本完整，肺泡腔內出血少，未見明顯滲出，間質輕度增厚，炎性細胞浸潤較少；OLV組右側肺組織肺泡結構破壞，肺泡腔內有少量滲出物，出血較少，炎性細胞浸潤較少，間質增厚；OLV組左側肺組織大部分肺泡結構消失，肺泡腔內有明顯滲出，出血多，，間質明顯增厚，炎性細胞浸潤較多； Cur組右、左側肺組織較OLV組右、左側肺組織損傷程度輕。 3TLV組內雙側肺組織W/D比、肺組織病理學損傷評分、MDA含量、SOD活性和Nrf2蛋白表達水平差異無統(tǒng)計學意義（P＞0.05）；與TLV組右、左肺比較，OLV組及Cur組右、左肺W/D比、肺組織病理學損傷評分、MDA含量、Nrf2蛋白表達水平升高，SOD活性降低（P＜0.05）；與OLV組右肺比較，OLV組左肺W/D比、肺組織病理學損傷評分、MDA含量、Nrf2蛋白表達水平升高，SOD活性降低（P＜0.05）。 4與OLV組右、左肺比較，Cur組右、左肺W/D比、肺組織病理學損傷評分、MDA含量下降，SOD活性、Nrf2蛋白表達水平升高（P＜0.05）；與Cur組右肺比較，Cur組左肺W/D比、肺組織病理學損傷評分、MDA含量、Nrf2蛋白表達水平升高，SOD活性降低（P＜0.05）。 結論：1單肺通氣3h可導致兔動脈血氧合指數(shù)下降，雙側肺組織病理性損傷且非通氣側肺組織較通氣側損傷嚴重。 2Nrf2/ARE信號通路在單肺通氣誘發(fā)的肺損傷中發(fā)揮肺保護作用。 3姜黃素預先給藥具有減輕單肺通氣所誘發(fā)的肺損傷作用，其機制與Nrf2蛋白表達上調有關。
[Abstract]:Objective: single lung ventilation (OLV) refers to the non ventilatory lung collapse in.OLV only using non operative side lung ventilation after thoracotomy, providing good visual field for operation and preventing two interpulmonary cross infection. However, the long duration of single lung ventilation can induce acute lung injury [1]. curcumin from Jiang Huanggen The active components extracted from the stem have shown that curcumin can reduce the lung injury induced by single lung ventilation, but the mechanism of action is not clear. The nuclear factor NF-E2 related factor 2 (Nrf2) is a key transcription factor for regulating oxidation / reduction reaction and inflammatory response in the body. The study found that Nrf2 plays an important role in acute lung injury, [3-4] This study is to evaluate the effect of curcumin on the expression of Nrf2 protein in lung tissue of rabbits with single lung ventilation, and to provide a reference for reducing the mechanism of lung injury induced by curcumin in single lung ventilation.
Methods: 24 healthy adult male New Zealand white rabbits, with a weight of 2.5~3.0kg, were provided by the experimental animal center of Hebei Medical University. The random digital table was used to randomly divide them into 3 groups (n=8): double lung ventilation group (group TLV), right lung single lung ventilation group (group OLV), and curcumin pre administration group (group Cur).
After the rabbit was weighed, 3% pentobarbital sodium 30mg/kg was injected into the right ear vein, and the free right femoral artery was used to monitor the average arterial pressure (MAP) and the intraoperative blood sample.2% hydrochloric acid lidocaine under local infiltration anesthesia. The ID3.0mm tracheal catheter was selected and the tracheal tube was inserted into the trachea in the TLV group after tracheotomy. The OLV group and the Cur group were placed in the right main bronchus for a single lung ventilation. By observing the changes in the right chest movement, hearing the double lung breathing sound and monitoring the airway pressure, the trachea was judged and adjusted. After the intubation was successful, the TLV group had two lung ventilation 3h, OLV group and Cur group with single lung ventilation 3H. The 3 groups all adopted the volume ventilation mode, FiO2100%, adjusted the ventilation parameters to maintain SpO2 > 90%. all the right lateral position, and the right internal jugular vein was injected with vecuronium 0.1mg/kg after the stable 10min, and the mechanical ventilation was connected to the ventilator after the spontaneous breathing disappeared. Group Cur and group Cur were first ventilated with single lung 50min, and then two lungs were ventilated by 10min (the ventilation parameters remained unchanged, the catheter was retreated to the endotracheal), and then the ventilation process was repeated. At the same time, the two groups all opened a size of about 1cm to the thoracic cavity at the left chest wall 5~6 intercostal, and used to observe the collapse and retention of the left lung tissue. The intermedium vein was given to vecuronium in the middle of the experiment. Ammonium 0.1mg/kg maintains muscle relaxation and maintains an intravenous infusion of 3% pentobarbital sodium 1ml if necessary to maintain a continuous infusion of sodium lactate solution at the rate of 10ml / kg-1 H-1 from the right internal jugular vein to supplement the fluid loss and maintain the stability of MAP during the experimental process. At the beginning of the ventilation (T0) and ventilation 1,2,3h (T1-3), the right femoral artery blood samples are collected. Blood gas analysis, determination of PaO2 and calculation of oxygenation index (OI), calculation formula: after the end of OI=PaO2/FiO2. ventilation, the animals were killed by rapid blood release of femoral artery, and the two lungs were taken to about 1cm * 1cm * 2cm size lung tissue and divided into two, and half of them were stored in 80oC refrigerator with liquid nitrogen cryopreservation, and the other half was fixed with 4oC4% polycondensation Formaldehyde Solution. The remaining lung tissue was weighed and placed in an oven (80 72h). The dry weight was calculated after constant weight and the wet / dry weight ratio (W/D ratio) of the lung was calculated.
The lung tissue was fixed in 4% polycondensation Formaldehyde Solution, routine dehydration, paraffin embedding, slice and HE staining. The pathological results were observed under optical microscope and the pathological damage score of lung tissue was observed. The expression of Nrf2 protein in lung tissue of rabbit lung tissue was detected by immunohistochemistry. The content of malondialdehyde (MDA) was detected by chemical colorimetry, and the superoxide was detected by chemical colorimetry. Dismutase (SOD) activity and Western blot method were used to detect the expression of Nrf2 protein in rabbit lung tissue.
Results: 1 compared with group TLV, OI in group OLV and Cur decreased (P < 0.05). Compared with T0, T2,3 OI decreased in OLV group and Cur group (P < 0.05).
In the 2 light microscope, the alveolar structure of bilateral lung tissues in group TLV was basically complete. There was little bleeding in the alveolus, no obvious exudation, mild thickening of interstitial tissue and less inflammatory cell infiltration. The alveolar structure in the right lung tissue of OLV group was destroyed, a small amount of exudation in the alveolus, less bleeding, less inflammatory cells infiltration, thickening of interstitial tissue, and most of the left lung tissues in group OLV. The alveolar structure disappeared, there were obvious exudation in the alveolus, more bleeding, more interstitial thickening and more inflammatory cell infiltration. Group Cur was right, left lung tissue was better than group OLV, and left lung tissue was less damaged.
There was no significant difference in the W/D ratio of bilateral lung tissues in the 3TLV group, the pathological damage score of the lung tissue, the MDA content, the SOD activity and the expression of Nrf2 protein (P > 0.05). Compared with the TLV group, the right of the TLV group, the left lung, the left lung W/D ratio, the pathological injury score of the lung tissue, the MDA content, the increase of the Nrf2 protein expression level and the decrease activity (0.05). Compared with the right lung in group OLV, the left lung W/D ratio, lung tissue pathological score, MDA content, Nrf2 protein expression level and SOD activity decreased in group OLV (P < 0.05).
4 in group OLV, right, left lung, Cur group right, left lung W/D ratio, lung tissue pathological damage score, MDA content decreased, SOD activity, Nrf2 protein expression level increased (P < 0.05); compared with Cur group right lung, Cur group left lung W/D ratio, lung tissue pathological injury score, MDA content, Nrf2 protein expression level increased and decreased activity (0.05).
Conclusion: 1 single lung ventilation 3H can lead to a decrease in arterial oxygenation index, bilateral lung tissue pathological damage and serious lung ventilation in non ventilated side.
2Nrf2/ARE signaling pathway plays a protective role in lung injury induced by one lung ventilation.
3 curcumin pretreatment can alleviate lung injury induced by one lung ventilation, and its mechanism is related to the up-regulated expression of Nrf2 protein.

【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R285.5

【參考文獻】

相關期刊論文 前10條

1 李慧婷;林文前;譚紅鷹;操隆輝;李靖;曾維安;;姜黃素減輕兔單肺通氣導致的肺損傷[J];中山大學學報(醫(yī)學科學版);2011年06期

2 劉文博;譚曉斌;賈曉斌;余嗣明;;基于Nrf2通路的中藥化學預防腫瘤的研究思路[J];中草藥;2011年07期

3 林文前;譚紅鷹;操隆輝;李慧婷;李靖;曾偉安;;單肺通氣誘發(fā)兔急性肺損傷模型的建立[J];中華麻醉學雜志;2011年03期

4 朱瑞芳;周敏;何建林;丁福云;余書勤;緒廣林;;姜黃素對大鼠油酸型急性肺損傷的保護作用[J];中國中藥雜志;2008年17期

5 游志堅;姚尚龍;李進;;三種兔單肺通氣模型的比較[J];中國實驗動物學報;2007年04期

6 游志堅;姚尚龍;袁茵;;單肺通氣后兩側肺損傷程度的比較[J];華中醫(yī)學雜志;2007年02期

7 連慶泉;李興旺;尚游;姚尚龍;馬利;金勝威;;Protective Effect of Curcumin on Endotoxin-induced Acute Lung Injury in Rats[J];華中科技大學學報(醫(yī)學英德文版);2006年06期

8 李瑜;李琳璋;王世端;曾因明;;黃芪對兔內毒素性急性肺損傷的保護作用[J];中國中西醫(yī)結合急救雜志;2006年06期

9 周剛;王繼峰;牛建昭;李銀生;陳偉濤;李澤華;劉同祥;;姜黃素抗肺纖維化大鼠細胞外基質過度形成的實驗研究[J];中國中藥雜志;2006年07期

10 王月蘭,姚尚龍;機械牽張導致肺上皮細胞損傷的信號轉導機制研究進展[J];國外醫(yī)學.麻醉學與復蘇分冊;2004年06期

相關博士學位論文 前1條

1 甯交琳;Nrf2在大鼠肢體爆炸傷后短暫低溫肺保護中的作用及機制[D];第三軍醫(yī)大學;2008年

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