多巴酚丁胺对脓毒症休克兔急性肺损伤的影响

发布时间：2018-01-30 14:03

本文关键词： 脓毒症休克急性肺损伤多巴酚丁胺水通道蛋白5 环磷酸腺苷　出处：《南京医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的通过建立脓毒症休克(septic shock)模型,观察水通道蛋白5(Aquaporin 5,AQP5)在脓毒症休克所致急性肺损伤(acute lung injury,ALI)时的变化规律,探讨不同剂量多巴酚丁胺对脓毒症休克所致ALI的保护作用及其可能的作用机制,从而为临床上ALI的治疗提供理论依据。方法采用盲肠结扎穿孔(cecal ligation and puncture,CLP)联合静脉注射内毒素(lipopolysaccharides,LPS)的方法制备脓毒症休克的动物模型,将70只新西兰雄性大白兔随机均分为假手术组(A组)、ALI模型组(B组)、多巴酚丁胺低剂量组(C组)、多巴酚丁胺中剂量组(D组)和多巴酚丁胺高剂量组(E组),各组分别于造模成功后3 h和6 h两个时相点各处死7只,留取肺组织标本,采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测各时相点肺组织中环磷酸腺苷(cyclic adenosine monophosphate,c AMP)的含量;蛋白质印迹法(Western-blot)及免疫组化法(Immunohistochemical analysis)检测AQP5的蛋白质水平;计算肺湿/干重(wet to dry weight,W/D)比值;光镜和电镜下观察各组兔肺组织的病理学改变,并对肺组织病理损伤程度进行评分。多组间差异比较采用单因素方差分析,组间两两比较采用LSD法。结果与A组相比,B组c AMP含量及AQP5蛋白水平在3 h及6 h均明显降低(P0.001),W/D比值则明显升高(P0.05)。与B组相比,C组c AMP及AQP5水平在6 h明显增加(P0.05),D、E组c AMP及AQP5水平在3 h、6 h明显增加(P0.05),E组W/D比值明显降低(P0.05)。与C、D组相比,E组c AMP及AQP5水平在3 h、6 h均有所增加(P0.01)。光镜及电镜下,B组肺组织的病理形态及超微结构损伤均较重,肺病理学评分明显升高(P0.01);C、D、E组肺的病理形态及超微结构损伤有所减轻,肺组织病理学评分明显降低(P0.01)。结论多巴酚丁胺对LPS诱导的ALI有一定程度的保护作用,且呈剂量依赖性关系,该作用机制可能与多巴酚丁胺提高肺组织c AMP水平,上调AQP5蛋白表达量有关。
[Abstract]:Objective to observe Aquaporin 5 by establishing septic shock model. The changes of AQP5 in acute lung injury-induced acute lung injury induced by septic shock. To investigate the protective effect of dobutamine at different doses on ALI induced by septic shock and its possible mechanism. Methods Cecal ligation and puncture was used for the treatment of ALI. CLP combined with intravenous injection of lipopolysaccharide lipopolysaccharide (LPSs) was used to establish septic shock model. Seventy New Zealand male rabbits were randomly divided into two groups: sham operation group (group A) and group B (model group) and dobutamine group (group C). Dobutamine medium dose group (group D) and dobutamine high dose group (group E) were killed in each group at 3 h and 6 h after successful modeling, respectively, and lung tissue specimens were collected. Enzyme-linked immunosorbent assay was determined by enzyme-linked immunosorbent assay (Elisa). The contents of cyclic adenosine monophosphate (adenosine) in lung tissue at different time points were determined by Elisa. The protein level of AQP5 was detected by Western blot and Immunohistochemical analysis. Lung wet / dry weight wet to dry weight W / D ratio was calculated. The pathological changes of lung tissue in each group were observed under light microscope and electron microscope, and the pathological injury degree of lung tissue was evaluated. The difference was compared by univariate analysis of variance (ANOVA). Results compared with group A, the content of c AMP and the level of AQP5 protein in group B were significantly lower than those in group A at 3 h and 6 h (P 0.001). Compared with group B, the levels of c AMP and AQP5 in group C were significantly higher than those in group B at 6 h. The levels of c AMP and AQP5 in group E were significantly increased at 3 h and 6 h, and the ratio of W / D in group E was significantly lower than that in group C (P 0.05), which was significantly lower than that in group C (P 0. 05). The levels of c AMP and AQP5 in group E were increased at 3 h and 6 h, respectively. The histopathological and ultrastructural damage of lung tissue in group B was serious under light microscope and electron microscope. The lung pathological score increased significantly (P 0.01). The pathological morphology and ultrastructure damage of the lung in group C were alleviated. Conclusion dobutamine can protect ALI induced by LPS in a dose-dependent manner. The mechanism may be related to the increase of c AMP level and the up-regulation of AQP5 protein expression in lung tissue by dobutamine.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R459.7

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