S-亚硝基-N-乙酰半胱氨酸对绿脓菌素感染鼠体内氧化作用的影响

发布时间：2018-09-19 10:10

【摘要】：目的探索绿脓菌素（PCN）对感染大鼠体内的氧化作用及其机制，进一步探讨S-亚硝基-N-乙酰半胱氨酸（SNAC）对此氧化作用的影响。方法 90只健康雄性Sprague—Dawley（SD）大鼠随机分为3组，对照组：大鼠腹腔注射1ml/100g体重0.9%氯化钠溶液，支气管接种生理盐水0.2ml；PCN组：大鼠腹腔注射1ml/100g体重0.9%氯化钠溶液，支气管接种PCN0.2ml；SNAC组：于感染前即刻给予SNAC（1ml/100g体重）干预，余同模型组。分别于6、18、30、48、72h处死大鼠，取支气管肺泡灌洗液（BALF）和肺组织。观察大鼠一般状态，肺组织形态学改变，计算BALF的细胞数，用试剂盒检测活性氧（ROS）含量，丙二醛（MDA）含量，超氧化物歧化酶（SOD）活性，westernblot检测SOD蛋白含量，试剂盒检测白介素-8（IL-8）含量。结果（1）大鼠的一般状态：PCN感染6h后大鼠出现精神萎靡、食欲不振、寒颤、发热、呼吸困难，感染30h最明显。分别于PCN感染后6、18、30、48、72h处死大鼠，观察肺组织，对照组大鼠肺外观呈粉红色，无明显异常改变；PCN组6h时可见肺组织表面散在点状出血；18～72h肺体积增大，肺组织暗红色，呈片状出血；SNAC组肺组织病变较PCN组明显减轻。（2）肺组织形态学变化：光镜下可见对照组肺组织结构清晰，肺泡间隔无增厚及炎性细胞浸润；PCN组肺组织肺泡壁结构完整性被破坏，肺泡间隔增厚，明显充血、水肿，肺泡内大量炎性细胞浸润，30h出现典型肺炎表现；与PCN组相比，SNAC组肺组织炎性表现明显减轻，肺泡壁结构完整性较好，充血、水肿减轻，炎性细胞浸润减少。（3）BALF细胞总数及中性粒细胞计数比较：3组大鼠PCN感染后6、18、30、48、72h BALF细胞总数、中性粒细胞计数比较，差异均有统计学意义（P＜0.05）；其中PCN组和SNAC组各时间点BALF细胞总数、中性粒细胞计数较对照组均增高，SNAC组各时间点BALF细胞总数、中性粒细胞计数较PCN组均降低，差异有统计学意义（P＜0.05）。（4）应用试剂盒测定结果显示：大鼠PCN感染后6、18、30、48、72h ROS含量、MDA含量、SOD活性、IL-8含量比较，差异均有统计学意义（P＜0.05）；其中PCN组和SNAC组各时间点ROS含量、MDA含量及IL-8含量较对照组均增高，SNAC组各时间点ROS含量、MDA含量及IL-8含量较PCN组均降低，PCN组和SNAC组各时间点SOD活性较对照组均降低，SNAC组各时间点SOD活性较PCN组均增高（P＜0.05）。（5）Western blot结果显示：大鼠PCN感染后6、18、30、48、72h SOD蛋白含量差异均有统计学意义（P＜0.05）；PCN组和SNAC组各时间点SOD蛋白含量较对照组降低，，SNAC组各时间点SOD蛋白含量较PCN组均增高（P＜0.05）,30h最明显。结论 1. PCN可以引起大鼠肺组织氧化损伤； 2. SNAC通过减少ROS保护PCN引起大鼠肺组织的氧化损伤。
[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.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R515

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