【作者】 何芳；

【导师】 彭再梅；

【作者基本信息】 中南大学 ， 临床医学， 2012， 硕士

【摘要】 目的：通过观察脂多糖(lipopolysaccharide, LPS)诱导的大鼠急性肺损伤(acute lung injury, ALI)中肺组织的病理变化、以及核转录因子-κB (Nuclear factor-KB,NF-KB)p65与基质金属蛋白酶-2(matrix metalloproteinase-2, MMP-2)在肺组织中的表达,探讨NF-κB信号转导通路及MMP-2在ALI中的作用；同时通过观察乌司他丁干预后ALI大鼠的肺组织病理、NF-κB p65及MMP-2的变化,探讨乌司他丁对脂多糖诱导的大鼠ALI是否具有保护作用以及可能的机制。方法：健康雄性SD大鼠45只,随机分为正常对照组,急性肺损伤组和乌司他丁干预组,每组15只。(1)正常对照组：尾静脉注射生理盐水(2ml/kg)。(2) ALI组：尾静脉注射脂多糖(5mg/kg)诱发ALI大鼠模型。(3)乌司他丁干预组：大鼠腹腔注射乌司他丁(100ku/kg),30分钟后尾静脉注射脂多糖(5mg/kg)。观察各组大鼠活动、精神等一般状态的变化,在各组尾静脉给药后4小时处死大鼠,并留取相应的肺组织和血液标本。抽取颈动脉血测定动脉血氧分压(arterial oxygen partial pressure, PaO2),取肺组织检测肺湿干重比(wet-to-dry weight ratio, W/D),肉眼观察肺组织大致改变,光镜下观察肺组织HE染色的病理改变,免疫组织化学法测定肺组织中NF-KBp65及MMP-2表达。结果：1.PaO2的变化：正常对照组大鼠Pa02波动在103.1mmHg左右；与正常对照组相比,ALI组大鼠Pa02显著降低(P<0.05)；乌司他丁干预组Pa02高于ALI组,但仍低于正常对照组水平,差异具有统计学意义(P<0.05)。2.肺组织W/D变化：正常对照组大鼠肺组织W/D波动在4.33左右；ALI组肺组织W/D与正常对照组比较显著增加(P<0.05)；乌司他丁干预组肺组织W/D较ALI组降低,但仍高于正常对照组水平。3.与正常对照组相比较,ALI组和乌司他丁干预组肺组织NF-κBp65及MMP-2的表达均显著增多(P<0.05)。乌司他丁干预组较ALI组NF-κBp65及MMP-2的表达低,差异有统计学意义(P<0.05)。结论：1.在脂多糖诱导的急性肺损伤大鼠肺组织中NF-κB p65及MMP-2的表达增高；2.乌司他丁可抑制急性肺损伤大鼠肺组织中NF-κB p65及MMP-2的表达；3.乌司他丁对脂多糖诱导的急性肺损伤大鼠可能具有保护作用。更多还原

【Abstract】 Objectives:To establish lipopolysaccharide (LPS)-induced acute lung injury (ALI) rat model and to investigate the pathological changes, the expression of nuclear factor KBp65(NF-κBp65) and matrix metalloproteinase-2(MMP-2) in the lung tissues of the ALI rat models. To study the effects of ulinastatinon the pathological changes, the expression of NF-KBp65and MMP-2in ALI rat, and determine the possible protection role ulinastatin in LPS-induced ALI rat.Methods:Forty-five healthy male SD rats were randomly divided into3groups, including blank control group, ALI group and ulinastatin group,with15rats in each group.(1) The blank control group was injected with saline(2ml/kg) through tail vein;(2) The ALI group was injected with LPS (5mg/kg) through tail vein to induce ALI.(3) The ulinastatin group was pretreated with ulinastatin (100ku/kg) through intraperitoneal injection and was injected with LPS (5mg/kg) through tail vein30minutesafter ulinastatin treatment. The general behavior of the rats from all groups was watched closely.4hours after the LPS injection, all the rats were sacrificed and lung tissue samples and arterial blood samples were collected correspondingly. The arterial blood sample was used for arterial oxygen partial pressure (PaO2) analysis. Different lobes of the lung tissue were used to determine lung wet to dry weight (W/D) ratio, to prepare for lung tissue for HE staining and to determine the expressions of NF-κBp65and MMP-2in lung tissue through immunohistochemistry analysis.Results:1. Changes in PaO2:PaO2values are around103.1mmHg in blank control group rats. Compared to blank control group, PaO2values in ALI group rats decreased dramatically (P<0.05), while PaO2values in ulinastatin group higher than those in ALI group, but still lower than those in blank control group, both differences are significant (P<0.05).2. Changes in lung tissue W/D ratio:lung tissue W/D ratios are around4.33in blank control group. Compared to blank control group, lung tissue W/D ratios in ALI group rats increased significantly (P<0.05), while lung tissue W/D ratios in ulinastatin group lower than those in ALI group, but still higher than those in blank control group, and both are significantly different (P<0.05).3. Compared with the blank control group, expressions of NF-κBp65and MMP-2in lung tissue were both significantly higher in ALI group and ulinastatin group (P<0.05), whilethose in ulinastatin group were lower than ALI group(P<0.05).Conclusion:1. The expressions of NF-κB p65and MMP-2are dramatically increased in LPS-induced ALI rats.2. Ulinastatin could inhibit the increase of NF-κB p65and MMP-2expressions in LPS-induced ALI rats.3. Ulinastatin can plays a protection role in LPS-induced ALI rat.更多还原