【摘要】：目的 探索綠膿菌素（PCN）對(duì)感染大鼠體內(nèi)的氧化作用及其機(jī)制，進(jìn)一步探討S-亞硝基-N-乙酰半胱氨酸（SNAC）對(duì)此氧化作用的影響。 方法 90只健康雄性Sprague—Dawley（SD）大鼠隨機(jī)分為3組，對(duì)照組：大鼠腹腔注射1ml/100g體重0.9%氯化鈉溶液，支氣管接種生理鹽水0.2ml；PCN組：大鼠腹腔注射1ml/100g體重0.9%氯化鈉溶液，支氣管接種PCN0.2ml；SNAC組：于感染前即刻給予SNAC（1ml/100g體重）干預(yù)，余同模型組。分別于6、18、30、48、72h處死大鼠，取支氣管肺泡灌洗液（BALF）和肺組織。觀察大鼠一般狀態(tài)，肺組織形態(tài)學(xué)改變，計(jì)算BALF的細(xì)胞數(shù)，用試劑盒檢測(cè)活性氧（ROS）含量，丙二醛（MDA）含量，超氧化物歧化酶（SOD）活性，westernblot檢測(cè)SOD蛋白含量，試劑盒檢測(cè)白介素-8（IL-8）含量。 結(jié)果 （1）大鼠的一般狀態(tài)：PCN感染6h后大鼠出現(xiàn)精神萎靡、食欲不振、寒顫、發(fā)熱、呼吸困難，感染30h最明顯。分別于PCN感染后6、18、30、48、72h處死大鼠，觀察肺組織，對(duì)照組大鼠肺外觀呈粉紅色，無(wú)明顯異常改變；PCN組6h時(shí)可見(jiàn)肺組織表面散在點(diǎn)狀出血；18～72h肺體積增大，肺組織暗紅色，呈片狀出血；SNAC組肺組織病變較PCN組明顯減輕。 （2）肺組織形態(tài)學(xué)變化：光鏡下可見(jiàn)對(duì)照組肺組織結(jié)構(gòu)清晰，肺泡間隔無(wú)增厚及炎性細(xì)胞浸潤(rùn)；PCN組肺組織肺泡壁結(jié)構(gòu)完整性被破壞，肺泡間隔增厚，明顯充血、水腫，肺泡內(nèi)大量炎性細(xì)胞浸潤(rùn)，30h出現(xiàn)典型肺炎表現(xiàn)；與PCN組相比，SNAC組肺組織炎性表現(xiàn)明顯減輕，肺泡壁結(jié)構(gòu)完整性較好，充血、水腫減輕，炎性細(xì)胞浸潤(rùn)減少。 （3）BALF細(xì)胞總數(shù)及中性粒細(xì)胞計(jì)數(shù)比較：3組大鼠PCN感染后6、18、30、48、72h BALF細(xì)胞總數(shù)、中性粒細(xì)胞計(jì)數(shù)比較，差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）；其中PCN組和SNAC組各時(shí)間點(diǎn)BALF細(xì)胞總數(shù)、中性粒細(xì)胞計(jì)數(shù)較對(duì)照組均增高，SNAC組各時(shí)間點(diǎn)BALF細(xì)胞總數(shù)、中性粒細(xì)胞計(jì)數(shù)較PCN組均降低，差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。 （4）應(yīng)用試劑盒測(cè)定結(jié)果顯示：大鼠PCN感染后6、18、30、48、72h ROS含量、MDA含量、SOD活性、IL-8含量比較，差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）；其中PCN組和SNAC組各時(shí)間點(diǎn)ROS含量、MDA含量及IL-8含量較對(duì)照組均增高，SNAC組各時(shí)間點(diǎn)ROS含量、MDA含量及IL-8含量較PCN組均降低，PCN組和SNAC組各時(shí)間點(diǎn)SOD活性較對(duì)照組均降低，SNAC組各時(shí)間點(diǎn)SOD活性較PCN組均增高（P＜0.05）。 （5）Western blot結(jié)果顯示：大鼠PCN感染后6、18、30、48、72h SOD蛋白含量差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）；PCN組和SNAC組各時(shí)間點(diǎn)SOD蛋白含量較對(duì)照組降低，，SNAC組各時(shí)間點(diǎn)SOD蛋白含量較PCN組均增高（P＜0.05）,30h最明顯。 結(jié)論 1. PCN可以引起大鼠肺組織氧化損傷； 2. SNAC通過(guò)減少ROS保護(hù)PCN引起大鼠肺組織的氧化損傷。
[Abstract]:Objective to investigate the oxidation of pyocyanin (PCN) in infected rats and its mechanism, and to explore the effect of Snitroso-N-acetylcysteine (SNAC) on this oxidation. Methods 90 healthy male Sprague-Dawley (SD) rats were randomly divided into three groups: the control group: rats were injected with 0.9% sodium chloride solution by intraperitoneal injection of 1ml/100g, the rats were given 0.9% sodium chloride solution by intraperitoneal injection of normal saline 0.2ml of bronchi, and the rats were injected with 0.9% sodium chloride solution by intraperitoneal injection of 1ml/100g. Bronchus inoculated PCN0.2ml;SNAC group: SNAC (1ml/100g body weight) was given immediately before infection. The rats were killed at 6: 18, 30, 48 and 72 hours, respectively. The bronchoalveolar lavage fluid (BALF) and lung tissue were taken. The changes of lung tissue morphology, the number of BALF cells, the content of reactive oxygen species (ROS), malondialdehyde (MDA), (SOD) activity of superoxide dismutase (SOD) and interleukin-8 (IL-8) were measured by Western blot. Results (1) the general state of rats was that the rats showed mental retardation, loss of appetite, chills, fever, dyspnea and infection for 30 h after 6 h infection. The lung tissues of the control group were observed at 72 h after PCN infection. The pulmonary appearance of the control group was pink, and the pulmonary tissue surface was scattered on the surface of the lung in the control group for 6 h. The lung volume was enlarged and the lung tissue was dark red at the end of 1872 hours after PCN infection. The pathological changes of lung tissue in SNAC group were significantly less than that in PCN group. (2) the lung histomorphology of the control group was clear under light microscope, and no thickening of alveolar septum and inflammatory cell infiltration were observed in the control group. In PCN group, the structural integrity of alveolar wall was destroyed, the alveolar septum thickened, hyperemia, edema, and a large number of inflammatory cells infiltrated in the alveoli appeared atypical pneumonia at 30 h, and the inflammatory manifestation of lung tissue in the SNAC group was significantly reduced compared with that in the PCN group. Alveolar wall structure integrity, hyperemia, edema, inflammatory cell infiltration decreased. (3) comparison of the total number of BALF cells and neutrophil count in rats of the 3 groups after PCN infection, the total number of BALF cells and neutrophil counts were compared at 72 h after PCN infection. The total number of BALF cells, neutrophil count and neutrophil count in PCN group and SNAC group were significantly higher than those in control group at each time point (P < 0. 05), and the neutrophil count was lower in SNAC group than in PCN group. The difference was statistically significant (P < 0. 05). (4) by using the kit. The results showed that the content of ROS and the activity of IL-8 were significantly higher than those of the control group (P < 0. 05). ROS content and IL-8 content in PCN group and SNAC group were higher than those in control group. ROS content and IL-8 content in SNAC group were lower than those in PCN group. SOD activity in PCN group and SNAC group was lower than that in control group at each time point. Compared with PCN group, the activity of SOD in each time point was higher than that in PCN group (P < 0. 05). The results showed that the content of SOD protein was significantly different from that in PCN group at 72 h after PCN infection (P < 0. 05). The content of SOD protein in PCN group and SNAC group was lower than that in control group at each time point. The content of SOD protein in SNAC group was significantly higher than that in PCN group at each time point (P < 0. 05). Conclusion 1. PCN can cause oxidative damage of lung tissue in rats. SNAC protects the lung tissue from oxidative damage induced by PCN by reducing ROS.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R515

【參考文獻(xiàn)】

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1 Subhankari Prasad Chakraborty;Santanu Kar Mahapatra;Sumanta Kumar Sahu;Sourav Chattopadhyay;Panchanan Pramanik;Somenath Roy;;Nitric oxide mediated Staphylococcus aureus pathogenesis and protective role of nanoconjugated vancomycin[J];Asian Pacific Journal of Tropical Biomedicine;2011年02期

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