不同钠离子浓度高渗盐复苏液对重度烫伤大鼠肝损伤的影响

发布时间：2018-03-07 17:31

  本文选题：高渗盐　切入点：烧伤　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:本实验通过采用600mmol/L、800mmol/L、1000mmol/L三种Na+浓度高渗钠盐液(HS)和乳酸格林钠液(LR)对重度烫伤大鼠进行液体复苏,从肝脏功能,肝组织细胞形态,炎症反应,氧化应激,炎症相关信号转导通道蛋白等方面探讨不同高渗钠盐液(HS)对大鼠重度烫伤后肝脏损伤的影响。方法:健康成年雌性SD大鼠104只,体重200-250g,随机分成五组:假伤组(8只)、其余为烫伤组包括LR组(24只)、600HS组(24只)、800HS组(24只)、1000HS组(24只)。大鼠麻醉后扎套管针尾静脉置管,剃去背部毛发,假伤组大鼠不予烫伤及补液,拟伤后直接取腹主动脉血及肝组织;其余各组均给予大鼠背部Ⅱ°至Ⅲ°开水烫伤,伤后分别使用LR复苏和三种不同HS复苏。烫伤组大鼠于补液后2h、8h和24h取腹主动脉血及肝组织。第一部分:不同钠离子浓度高渗盐复苏液对重度烫伤大鼠肝功能损伤及肝细胞损伤的影响取出五组大鼠的一部分肝脏组织及血浆,使用自动生化仪检测血浆中ALT及AST含量,肝脏组织在使用10%甲醛固定后使用HE染色法染色观察组织细胞形态。第二部分:不同钠离子浓度高渗盐复苏液对重度烫伤大鼠肝脏氧化损伤及炎症因子的影响取出五组大鼠的一部分肝脏组织及血浆,比色法对肝组织进行丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性测定。酶联免疫吸附法(ELISA)测量血浆中肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)以及高迁移率族蛋白B1(HMGB-1)。将取得数据进行统计分析。第三部分:不同钠离子浓度高渗盐复苏液对重度烫伤大鼠肝脏组织p38MAPK及JNK通道活化的影响使用蛋白质印迹法(Western Blot)测量五组大鼠肝脏组织中p38MAPK以及JNK通道磷酸化蛋白比值并进行统计学分析。结果:第一部分与假伤组相比,伤后各时间点,其余各组ALT、AST均明显升高(P0.01);与LR组相比,伤后各时间点,600HS组ALT、AST均明显降低,(P0.01),而800HS组及1000HS组ALT、AST在伤后2、8小时未见显著变化(P0.05),在伤后24小时800HS组及1000HS组ALT、AST均明显降低(P0.01);与600HS组相比,800HS组及1000HS组ALT、AST在伤后2、8小时明显升高(P0.01),在伤后24小时,800HS组及1000HS组ALT、AST未见显著差异(P0.05)。肝脏组织HE染色显示:伤后各时间点,LR组大鼠肝细胞可见明显脂肪样变,部分可见细胞水肿;伤后2、8小时HS组大鼠随高渗盐浓度升高,上述病理变化增多;伤后24小时,三组HS组大鼠肝脏病理变化无明显差异。第二部分与假伤组相比,伤后各时间点,LR组、800HS组和1000HS组TNF-α与IL-1β均明显升高(P0.01),而600HS组未见明显变化(P0.05);与LR组相比,伤后各时间点600HS组TNF-α与IL-1β均明显降低(P0.01),800HS组和1000HS组未见显著变化(P0.05)。伤后2小时,五组HMGB-1无显著差异(P0.05),伤后8、24小时,与假伤组相比,LR组、800HS组和1000HS组HMGB-1均明显升高(P0.01),而600HS组未见明显差异(P0.05);与LR组相比,各HS组HMGB-1均明显降低(P0.01);与600HS组相比,800HS组及1000HS组HMGB-1明显升高(P0.01)。与假伤组相比,伤后各时间点其余各组MDA含量明显升高SOD活性明显降低(P0.01);与LR组相比,伤后各时间点600HS组MDA含量明显降低SOD活性明显升高(P0.01),800HS组、1000HS组未见显著变化(P0.05);与600HS组相比,伤后各时间点800HS组及1000HS组MDA含量明显升高SOD活性明显降低(P0.01)。第三部分肝脏组织p38MAPK以及JNK通道磷酸化蛋白比值显示:LR组和各HS组在伤后2h、8h及24h,样本肝脏组织p38MAPK及JNK通道的活性表达均比假伤组显著升高(P0.01);而LR组、800HS组及1000HS组p38MAPK及JNK通道活性表达均显著高于600HS组(P0.01),而800HS组及1000HS组p38MAPK及JNK通道活性与LR组无差异(P0.05)。结论:(1)与伤后使用LR相比,使用600mmol/L钠离子浓度的高渗盐溶液进行液体复苏能减轻但不终止大鼠的肝功能失常及肝细胞形态结构的损伤,但是随着Na+浓度升高达到800及1000mmol/L后,效果降低。(2)与伤后使用LR相比,使用600mmol/L钠离子浓度的高渗盐溶液进行液体复苏能减轻并部分终止重度烫伤大鼠伤后肝脏的氧化损伤以及炎症反应发生,但是随着Na+浓度升高,达到800及1000mmol/L后,效果降低。(3)与伤后使用LR相比,使用600mmol/L钠离子浓度的高渗盐溶液进行液体复苏能减轻但不终止肝脏组织中p38MAPK和JNK通道的激活表达,但是随着Na+浓度升高,达到800及1000mmol/L后,效果降低。
[Abstract]:Objective: this experiment by using 600mmol/L, 800mmol/L, 1000mmol/L three Na+ concentration of hypertonic salt solution (HS) and Green (LR) lactic acid liquid sodium on Severe Scalded Rats by fluid resuscitation from the liver function, liver cell morphology, inflammation, oxidative stress, inflammation related signaling channel protein of different hypertonic salt solution (HS) of rats after severe burns after liver injury. Methods: 104 healthy adult female SD rats, weighing 200-250g, were randomly divided into five groups: sham injury group (8 rats), the rest of the scald group including LR group (24 rats), 600HS group (24), 800HS group (24 rats), 1000HS group (24 rats). Rats were anesthetized with intravenous trocar catheter, shaved back hair, the sham injury group rats with scald and rehydration, to directly after the injury abdominal aorta blood and liver tissue; the other groups were given back to the boiling water second-degree third degree in rats burns, LR respectively after injury recovery And three different HS recovery. Scald rats in group 8h and 24h after 2H infusion, abdominal aorta blood and liver tissue. The first part: different concentration of sodium ions of hypertonic saline resuscitation fluids on severe burn injury and liver function of liver cell injury in rats out of the five groups of rats a portion of the liver tissue and plasma the use of ALT, and the content of AST automatic biochemical detection of plasma and liver tissue using HE staining to observe cell morphology staining in 10% formalin fixed. The second part: the different concentration of sodium ions of hypertonic saline resuscitation fluid effects on liver oxidative damage and inflammatory factors in rats with severe burn out of the five groups of rats a portion of the liver tissue and plasma, colorimetry of liver tissue malondialdehyde (MDA) content, superoxide dismutase (SOD) activity assay. Enzyme linked immunosorbent assay (ELISA) in the measurement of plasma tumor necrosis factor alpha (TNF- alpha), interleukin 1 beta (IL-1 beta) to And the high mobility group protein B1 (HMGB-1). The obtained data were statistically analyzed. The third part: the sodium ion concentration of hypertonic saline resuscitation fluid using the method of severe scald rat liver tissue p38MAPK and JNK channel activation by Western blot (Western Blot) in liver tissue were measured in five groups of rats in p38MAPK and JNK channel phosphate the ratio of protein and analyzed statistically. Results: the first part compared with the sham injury group at each time point, the rest of groups ALT, AST were significantly increased (P0.01); compared with the LR group at each time point in group 600HS, ALT, AST were significantly lower (P0.01), and 800HS group 1000HS ALT AST group, there was no significant changes in 2,8 hours after injury (P0.05), 24 hours after injury in 800HS group and 1000HS group ALT, AST were significantly decreased (P0.01); compared with 600HS group, 800HS group and 1000HS group ALT, AST in 2,8 hours after injury was significantly increased (P0.01), after injury 24 hours, 800HS group 鍙，

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