脓毒症性急性肺损伤大鼠HMGB1表达及乌司他丁干预的实验研究

发布时间：2018-04-29 06:15

本文选题：急性肺损伤 + 乌司他丁　；参考：《第二军医大学》2014年硕士论文

【摘要】：急性肺损伤（Acute Lung Injury，ALI）是继发于严重创伤、感染、休克和大手术后，以呼吸窘迫和顽固性低氧血症为临床特征的综合征，急性肺损伤是多脏器功能障碍综合征(multiple organ dysfunction syndrome, MODS)在肺部的突出表现，急性呼吸窘迫综合征（Acute respiratorydistress syndrome，ARDS）是其最严重的表现形式。急性肺肺损伤的发病率为每年38/10万，近年来有逐渐增高的趋势，尽管采取了很多措施，急性肺损伤的死亡率仍高达32%-50%，高迁移率族蛋白B1（Highmobility group box1protein，HMGB1）是一种晚期炎症介质，同时也是一种促炎细胞因子，能够介导炎症反应并保持级联扩大作用，其生物学特性跟多种疾病的发病机制有很高的关联性，越来越多的研究证实，炎性因子在急性肺损伤的发生发展中起着重要作用；乌司他丁（Ulinastatin，UTI）为一种蛋白酶抑制剂，早期用于急性胰腺炎的治疗，有研究表明乌司他丁可以抑制急性肺损伤/急性呼吸窘迫综合征的发生发展，降低急性肺损伤/急性呼吸窘迫综合征的病死率，但目前对于乌司他丁的作用机制尚未完全明确，本实验通过乌司他丁干预脓毒症性急性肺损伤大鼠HMGB1的表达及水平变化，探讨乌司他丁减轻急性肺损伤程度、降低疾病死亡率可能的机制。第一部分脓毒症性急性肺损伤大鼠7天生存率观察目的通过盲肠结扎穿刺（CLP）手术法致大鼠脓毒症性急性肺损伤（ALI），观察各组大鼠7天生存率，探讨乌司他丁有无改善脓毒症性急性肺损伤大鼠生存率的作用。方法雄性健康SD大鼠30只随机分入CLP组，乌司他丁组，假手术组（对照组），每组10只，CLP组大鼠行CLP手术干预，乌司他丁组CLP手术+乌司他丁干预，假手术组仅做腹部开关手术；观察各组大鼠7天生存率。结果CLP组大鼠7天生存率50%，乌司他丁组7天生存率70%，假手术组（对照组）大鼠7天生存率100%，CLP组与乌司他丁组大鼠7天生存率有明显差异（P＜0.05）。结论乌司他丁可以明显改善脓毒症性急性肺损伤大鼠7天生存率，早期干预脓毒症急性肺损伤可能对改善疾病总体预后具有重要作用。第二部分脓毒症性急性肺损伤大鼠HMGB1表达及乌司他丁干预的实验研究目的通过CLP手术致大鼠脓毒症性急性肺损伤，乌司他丁干预急性肺损伤，研究乌司他丁对高迁移率族蛋白B1（HMGB1）的表达变化，探讨乌司他丁治疗脓毒症性急性肺损伤的分子生物学机制。方法雄性健康SD大鼠120只随机分入CLP组、乌司他丁组、假手术组；CLP组、乌司他丁组各40只，，组内分6h组10只、12h组10只、24h组10只、48h组10只；假手术组40只,组内分组同前。乌司他丁组大鼠于CLP手术30分钟后按10万U/kg由大鼠尾静脉注射乌司他丁，CLP组及假手术组注射等量生理盐水，各组大鼠在6h、12h、24h、48h设定时间点留取相关标本进行相关指标检测。结果假手术组（对照组）各时点肺组织HMGB1mRNA表达量均显著低于CLP组、乌司他丁组对应时点肺组织HMGB1mRNA表达量（P0.05）；乌司他丁组HMGB1mRNA表达量在12h、24h、48h点均显著低于CLP组对应时点（P0.05）；乌司他丁组各时点肺组织中HMGB1含量均显著低于CLP组中对应时点HMGB1含量（P0.05）。CLP组大鼠血浆中各时点HMGB1含量明显高于假手术组（对照组）、乌司他丁组对应时点表达含量（P0.05）；乌司他丁组各时点BALF中HMGB1含量均显著低于CLP组中对应时点HMGB1含量（P0.05）。结论HMGB1参与了脓毒症性急性肺损伤的发生发展过程，乌司他丁通过下调HMGB1的表达达到减轻急性肺损伤程度，提高大鼠生存率的作用。
[Abstract]:Acute lung injury (Acute Lung Injury, ALI) is a syndrome secondary to severe trauma, infection, shock, and major surgery, with respiratory distress and intractable hypoxemia as a clinical feature. Acute lung injury is a prominent manifestation of multiple organ dysfunction syndrome (multiple organ dysfunction syndrome, MODS) in the lungs, and acute respiratory distress syndrome Acute respiratorydistress syndrome (ARDS) is the most serious form of expression. The incidence of acute lung and lung injury is 38/10 million each year. In recent years, there is a tendency to increase gradually. Although many measures have been taken, the mortality of acute lung injury is still as high as 32%-50%, and high mobility group protein B1 (Highmobility group box1protein, HMGB1) is a kind of disease. Advanced inflammatory mediators, also a pro-inflammatory cytokine, can mediate inflammatory reactions and maintain cascade expansion, and their biological characteristics are highly associated with the pathogenesis of various diseases. More and more studies have shown that inflammatory factors play an important role in the development of acute lung injury; Ulinastatin UTI) is a protease inhibitor, early in the treatment of acute pancreatitis. Studies have shown that Ulinastatin can inhibit the development of acute lung injury / acute respiratory distress syndrome and reduce the mortality of acute lung injury / acute respiratory distress syndrome. However, the mechanism of ulinastatin is not yet fully defined. The possible mechanism of Ulinastatin to reduce the degree of acute lung injury and reduce the mortality of the acute lung injury of septic rats was examined through the intervention of ulinastatin in the expression and level of HMGB1 in the rats with acute lung injury.
Part 7 survival rate of septic acute lung injury in rats
Objective To observe the effect of ulinastatin on the survival rate of rats with acute septic lung injury by using the method of cecal ligation (CLP) to induce septic acute lung injury (ALI) in rats, and to observe the 7 natural survival rate of rats in each group.
Methods 30 male healthy SD rats were randomly divided into group CLP, ulinastatin group, sham operation group (control group), 10 in each group. Group CLP rats were treated with CLP operation, ulinastatin group CLP operation plus Ulinastatin, and sham operation group only underwent abdominal switching operation, and the 7 day survival rate of rats in each group was observed.
Results the 7 natural survival rate was 50% in the CLP group, 70% in the Ulinastatin group 7, 100% in the sham operation group (control group) and 100% in the rats in the control group. The survival rate of 7 in the CLP group and the Ulinastatin group was significantly different (P < 0.05).
Conclusion Ulinastatin can significantly improve the 7 born rate of septic acute lung injury in rats. Early intervention in acute lung injury of sepsis may play an important role in improving the overall prognosis of the disease.
The second part is the expression of HMGB1 in septic acute lung injury rats and the intervention of ulinastatin.
Objective to study the effect of ulinastatin on the expression of high mobility group protein B1 (HMGB1) and the molecular mechanism of ulinastatin in the treatment of acute lung injury of sepsis by CLP operation in rats with acute lung injury caused by sepsis.
Methods 120 male healthy SD rats were randomly divided into group CLP, ulinastatin group, sham operation group, CLP group and ulinastatin group 40, group 6h, 10 in group 6h, 10 in group 12h, 10 in group 48h, 40 in group 48h, and 40 in sham operation group, and in group of ulinastatin group at 100 thousand U/kg after 30 minutes of CLP hand, and intravenous injection of ulinastin by 100 thousand U/kg in rat tail veins at 100 thousand U/kg after 100 thousand U/kg The rats in group CLP and sham operation were injected with the same amount of normal saline. The rats in each group were given relevant samples at 6h, 12h, 24h, 48h setting time, and related indexes were detected.
Results the expression of HMGB1mRNA in the lung tissue at all time points in the sham operation group was significantly lower than that in the CLP group. The expression of HMGB1mRNA in the lung tissue at the time point in the Ulinastatin group (P0.05), the HMGB1mRNA expression of the Ulinastatin group in 12h, 24h, 48h were significantly lower than that in the CLP group at the corresponding time point (P0.05), and the HMGB1 content in the lung tissues at each time point in the Ulinastatin group was obvious. The content of HMGB1 in the plasma at the corresponding time point HMGB1 content (P0.05) in group.CLP was significantly higher than that in the sham group (control group) at the corresponding time point of the corresponding time point (P0.05) in group.CLP (P0.05). The content of the BALF in the Ulinastatin group was (P0.05), and the HMGB1 content in the BALF at each time point in the Ulinastatin group was significantly lower than the HMGB1 content at the corresponding time point at the CLP group (P0.05).
Conclusion HMGB1 is involved in the development and development of acute lung injury in sepsis. Ulinastatin can reduce the degree of acute lung injury and improve the survival rate of rats by down regulating the expression of HMGB1.

【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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