乌司他丁对脓毒症大鼠心肌能量代谢相关基因表达的影响

发布时间：2018-04-28 11:46

  本文选题：脓毒症 + 乌司他丁　； 参考：《福建医科大学》2016年硕士论文

【摘要】：脓毒症是感染引起的全身炎症反应综合征(SIRS)。心功能障碍常出现在严重脓毒症和脓毒症休克中,但其发生发展的机制尚未完全明了。其中心肌能量代谢障碍是心肌细胞损伤的始动环节之一,是引起和促进心功能障碍发生、发展的重要因素。针对脓毒症和脓毒症休克的诊疗方法一直在不断地寻找改良和运用,除了经典抗感染治疗、液体复苏、保护性通气等,一些辅助治疗也越来越多的被关注、被研究与应用,包括糖皮质激素的应用、免疫球蛋白的使用、抗凝剂的应用、血糖控制、他汀类药物的应用,以及乌司他丁的使用等。乌司他丁(UTI)是从人尿中提取的精制的蛋白酶抑制剂,可抑制胰蛋白酶、弹性蛋白酶、纤溶酶等蛋白水解酶以及透明质酸酶、淀粉酶、脂肪酶等糖类和脂类水解酶。目前国内外关于乌司他丁对脓毒症心肌能量代谢的研究较少,特别是几乎无相关的基因水平上的研究。本实验将在基因水平上探究乌司他丁对脓毒症心肌能量代谢的影响及推断其可能的作用机制,为临床上合理用药提供依据。方法:45只雄性Wistar大鼠随机分为对照组、脓毒症组和乌司他丁组。采用盲肠结扎穿孔术(CLP)制作脓毒症大鼠模型;对照组仅开腹、关腹,不行CLP。乌司他丁组于制模前1 h肌肉注射乌司他丁10万u/kg;术后每隔8个小时重复给药,脓毒症组及假手术组肌注平衡液5 ml/kg。于CLP术后24 h在氯胺酮腹腔麻醉下迅速开腹、取心脏,并立即置入液氮中,-70℃保存,以待提取RNA。运用RT2profile PCR阵列技术进行检测,分析比较脓毒症组、乌司他丁组与对照组大鼠心肌能量代谢相关基因表达的变化,并比较分析脓毒症组和乌司他丁组差异表达基因之间的不同。结果:脓毒症组和乌司他丁组均出现了心肌能量代谢相关基因表达变化。与对照组相比,乌司他丁组中有11个基因出现差异表达,其中有9个表达下调(Atp12a、Atp4a、Atp5d、Atp6v0a2、Atp6v1g3、Cox4i2、Cyc1、Slc25a10、Slc25a15、),2个表达上调(Atp6v0d2、Atp6v1e2);脓毒症组有5个基因出现差异表达,其中有2个表达下调(Atp6v1g3、Ucp1),3个表达上调(Atp12a、Atp4a、Ucp3)。乌司他丁组和脓毒症组表达差异的基因有所不同,乌司他丁有调节作用的基因有13个。涉及的基因编码的蛋白主要与呼吸链酶复合物V(ATP合酶)、酶复合体IV(细胞色素c氧化酶)、酶复合体III(辅酶Q-细胞色素c还原酶)和能量代谢相关的辅助蛋白有关。结论:脓毒症时机体出现了心肌能量代谢方面的改变,乌司他丁对脓毒症诱发的心肌能量代谢障碍起到一定改善作用。
[Abstract]:Sepsis is a systemic inflammatory response syndrome caused by infection. Cardiac dysfunction often occurs in severe sepsis and septic shock, but the mechanism of its development has not been fully understood. Among them, myocardial energy metabolism disorder is one of the initiation links of myocardial cell injury, which is an important factor to cause and promote the occurrence and development of cardiac dysfunction. The methods of diagnosis and treatment for sepsis and septic shock have been constantly looking for improvement and application. In addition to classical antiinfective therapy, fluid resuscitation, protective ventilation, and so on, some adjuvant treatments have been paid more and more attention to, studied and applied. These include the use of glucocorticoids, immunoglobulin, anticoagulants, blood glucose control, statins, and ulinastatin. Ulinastatin UTI is a refined protease inhibitor extracted from human urine. It can inhibit proteolytic enzymes such as trypsin, elastase, fibrinolytic enzyme, hyaluronidase, amylase, lipase and so on. There are few studies on the energy metabolism of sepsis myocardium at home and abroad, especially at the level of almost no related genes. This study will explore the effect of ulinastatin on myocardial energy metabolism in sepsis and its possible mechanism at the gene level. Methods 45 male Wistar rats were randomly divided into control group, septic group and ulinastatin group. The sepsis rat model was made by cecal ligation and perforation, while the control group was only open and closed abdomen, but not CLP. Ulinastatin group was intramuscularly injected with ulinastatin 100000 u / kg one hour before model making, and repeated administration every 8 hours after operation. The sepsis group and sham operation group were injected intramuscularly with a balanced solution of 5 ml / kg. After 24 hours after CLP, the heart was quickly opened under ketamine abdominal anesthesia, and the heart was stored in liquid nitrogen at -70 鈩，

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