高压电烧伤对大鼠血清β-TG和PF-4的影响及乌斯他丁的干预作用

发布时间：2018-11-18 07:36

【摘要】：目的:高压电烧伤除即刻性电热损伤外,还存在着原因未明的渐进性损伤。研究表明,微循环障碍在电烧伤后的渐进性损伤发生和发展过程中起关键作用,并发现电伤后的微循环障碍主要由微血管损伤及微血流障碍所致。在影响微循环血流动力学和流变学指标中,血小板流变行为异常起重要作用。血小板是血液的主要成分之一,是影响微循环血流动力学及流变学的重要因素。高压电烧伤通过激发血小板活化,使血小板聚集性增加、黏附性增强、促释放反应,从而加重微循环障碍。但血小板流变行为异常的分子机制目前尚不清楚。本研究通过检测大鼠高压电烧伤后早期血清β-血小板球蛋白(β-TG)及血小板因子4(PF-4)含量,并采用乌司他丁(UTI)进行干预,旨在进一步探索高压电烧伤后血小板流变行为异常的相关机制和防治方法。方法:1分组健康成年雄性SD大鼠240只,随机分为假高压电烧伤组(假伤组)、高压电烧伤组(电伤组)、高压电烧伤复合盐水治疗组(盐水组)、高压电烧伤复合UTI治疗组(UTI组),每组各60只。每组又按观察时间分为伤前15min、伤后5min、伤后1h、2h、4h、8h六个时相组,每时相组10只。2模型制作大鼠腹部备皮及消毒。用10%水合氯醛溶液腹腔注射麻醉大鼠(3ml/kg),麻醉成功后,将大鼠仰卧固定于专用电击实验台上,将两个1cm×1cm电极片分别固定于大鼠的左上肢(电流入口)、右下肢(电流出口)。连接实验变压器和调压器电线,接通电源,调节调压器使升压器输出电压至2k V,接通升压器输出电源,使高压电流通过大鼠,电击3s,模型制作成功。假伤组只连电线不通电致假伤。电伤5min内,UTI组腹腔注射1%UTI(按2×104u/kg即2ml/kg给药),盐水组腹腔内注射2ml/kg生理盐水。3指标检测:对模型大鼠开胸暴露心脏,直视下心脏抽血6ml,置于无菌塑料试管中,静置30min,待血清析出后上离心机,以3000转/min离心10min,取上清液置于Eppendorf管中在-70℃条件下保存。采用ELISA双夹心抗体法,分别检测每组大鼠六个时相组血清β-TG、PF-4含量。4统计处理:采用SPSS 21.0统计软件,行两因素析因设计的方差分析,多重比较采用LSD法t检验。以P0.05为显著性检验水准。结果:1大鼠血清β-TG含量变化电伤组血清β-TG含量的平均水平高于假伤组(F=172.186,P0.01);电伤组血清β-TG含量受电伤后时间变量影响(F=32.164,P0.01),在电伤后1h、2h、4h和8h时高于伤前值(P值均小于0.01)。UTI组血清β-TG含量的平均水平低于盐水组(F=21.295,P0.01);UTI血清β-TG含量受电伤后用药时间变量影响(F=109.115,P0.01),在电伤后1h和4h时低于盐水组(P值均小于0.01)。2大鼠血清PF-4含量变化电伤组血清PF-4含量的平均水平高于假伤组(F=29.108,P0.01);电伤组血清PF-4含量受电伤后时间变量影响(F=6.943,P0.01),在电伤后2h和8h时高于伤前值(P值均小于0.01)。UTI组血清PF-4含量的平均水平低于盐水组(F=37.874,P0.01);UTI血清PF-4含量受电伤后用药时间变量影响(F=15.026,P0.01),在电伤后1h、2h、4h、8h时低于盐水组(P值均小于0.01)。结论:高压电烧伤可促进大鼠血小板释放反应,引起大鼠高压电烧伤早期血清β-TG和PF-4含量升高。UTI则抑制大鼠血小板释放反应,使大鼠高压电烧伤后血浆β-TG和PF-4含量降低,从而改善血小板流变行为。
[Abstract]:Objective: In addition to the immediate heating injury of high-voltage electric burn, there is an unknown progressive injury. The study showed that the microcirculatory disturbance played a key role in the occurrence and development of progressive injury after electric burn, and found that the microcirculatory disturbance after electric burn was mainly caused by microvessel injury and micro-blood flow. The changes of platelet rheological behavior play an important role in the hemodynamic and rheology of the microcirculation. The platelet is one of the main components of the blood, which is an important factor that affects the hemodynamics and the rheology of the microcirculation. The high-voltage electric burn can stimulate the activation of the platelet, increase the aggregation of the platelet, enhance the adhesion and release the reaction, thereby increasing the microcirculation disturbance. But the molecular mechanism of the abnormal platelet-rheological behavior is not yet clear. In this study, the content of platelet factor 4 (PF-4) and the content of platelet factor 4 (PF-4) were measured in the early stage of high-voltage burn in rats, and the intervention of Ulinastatin (UTI) was used to further explore the related mechanisms and methods to prevent the abnormal changes of platelet rheological behavior after high-voltage burn. Methods: 240 healthy adult male SD rats were randomly divided into pseudo-high-voltage burn group (sham-wound group), high-voltage electric burn group (electric injury group), high-voltage burn composite saline treatment group (saline group), high-voltage burn composite UTI treatment group (UTI group), each group of 60 rats. In each group, the observation time was divided into 15 minutes before the injury, 5 minutes after the injury, 1 h, 2 h, 4 h and 8 h after the injury. Anesthetized rats (3ml/ kg) were injected intraperitoneally with 10% water-chloral solution, and after the anesthesia was successful, the rats were fixed on a special electric shock test bench, and the two 1cm-1cm electrode pieces were fixed to the left upper limb (current inlet) and the right lower limb (current outlet) of the rat respectively. Connect the test transformer and voltage regulator wire, turn on the power supply, adjust the voltage regulator to make the booster output voltage to 2k V, turn on the booster output power supply, and make the high-voltage current pass through the rat and the electric shock 3s, and the model is made to be successful. The false-wound group is not only connected with a wire to cause a false injury. In 5 minutes of electroinjury, 1% of UTI (2/ 104u/ kg, i.e. 2ml/ kg) was injected into the abdominal cavity of the UTI group, and 2ml/ kg of normal saline was injected intraperitoneally in the saline group. The solution was centrifuged at 3000 rpm for 10 min, and the supernatant was placed in the Eppendorf tube at-70.degree. C. for 10 min. The serum levels of triglyceride, PF-4 and the content of PF-4 in six groups of rats in each group were respectively detected by ELISA and double-sandwich antibody method. The results of statistical analysis were as follows: SPSS 2.0 statistical software and two-factor analysis of variance of the design were performed, and the multiple comparisons were tested by means of LSD. The level of the test was significantly higher than that of P 0.05. Results: The level of serum triglyceride-TG in the serum of the rats was higher than that of the sham-wound group (F = 172.186, P 0.01), and the serum level of TG-TG in the electroinjury group was affected by the time-dependent variable after electroinjury (F = 32.164, P0.01). At 4h and 8h, the pre-injury value (P-value was less than 0.01). The level of serum triglyceride-TG in the UTI group was lower than that of the saline group (F = 21.295, P0.01); the content of the serum triglyceride-TG in the UTI group was affected by the time-dependent variable after electroinjury (F = 109.115, P0.01). The mean level of PF-4 in the serum of the rats was higher than that of the sham-wound group (F = 29. 108, P 0.01), and the serum PF-4 content in the electroinjury group was affected by the time-dependent variable after electroinjury (F = 6.943, P0.01). The pre-injury value (P-value was less than 0.01) at 2h and 8h after electroinjury. The mean level of PF-4 in the UTI group was lower than that of the saline group (F = 37. 874, P0.01); the content of PF-4 in the UTI group was affected by the time-dependent variable after electroinjury (F = 15.026, P0.01), and was lower than that of the saline group at 1h, 2h, 4h, and 8h after electric injury (P <0.01). Conclusion: High-voltage burn can promote the release of platelet in rats, and raise the level of serum level-TG and PF-4 in the early stage of high-voltage burn in rats. UTI inhibited platelet-release in rats, and decreased the content of plasma-TG and PF-4 after high-voltage burn in rats, thus improving the rheological behavior of platelets.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R647

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