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目的观察重症失血性休克及复苏过程中大鼠小肠及血清中血管黏附蛋白1(VAP-1)的表达和活性变化,探讨抑制其功能对休克预后的影响。方法将实验大鼠随机分为假手术组、休克组、休克复苏组、复苏对照组和复苏实验组,每组10只。建立重症失血性休克及复苏大鼠模型,采用蛋白质印迹和real-time RT-PCR法检测假手术组、休克组、休克复苏组休克前、休克1h、复苏1h时小肠组织中VAP-1表达及其编码基因含量,ELISA法检测血清中VAP-1含量及其活性。复苏实验组加用20mg/kg 2-溴乙胺,复苏对照组加用1mL/kg生理盐水,比较两组复苏后的血压、复苏24h后小肠黏膜损伤情况(Chiu’s评分)和小肠黏膜上皮细胞凋亡情况(TUNEL),并比较大鼠24h生存率。结果重症失血性休克组大鼠小肠组织VAP-1蛋白水平以及其编码基因mRNA水平、血清中VAP-1含量及其活性均较假手术组升高(均P<0.05),复苏后上述各值均有所下降,但仍高于假手术组。相对于生理盐水对照组,休克复苏后1h和24h使用20mg/kg 2-溴乙胺的复苏实验组大鼠血压升高(P值分别为0.010和0.039),且复苏实验组Chiu’s评分及肠黏膜上皮细胞凋亡指数均低于生理盐水复苏对照组(P值分别为0.022和0.002),休克大鼠24h生存率也高于复苏对照组(90%vs 60%)。结论重症失血性休克能使大鼠VAP-1的表达、活性增高,而液体复苏能减轻这种增高;抑制其活性能改善重症失血性休克及复苏后的低血压以及小肠黏膜的损伤和细胞凋亡,提高大鼠24h生存率。
Objective To observe the expression and activity of VAP-1 in small intestine and serum during severe hemorrhagic shock and resuscitation in rats, and to explore the effect of suppressing its function on the prognosis of shock. Methods The experimental rats were randomly divided into sham operation group, shock group, shock resuscitation group, resuscitation control group and resuscitation experimental group, with 10 rats in each group. The rat model of severe hemorrhagic shock and resuscitation was established. The expression of VAP-1 in small intestine tissue of sham operation group, shock group and shock resuscitation group before shock, 1 hour after shock and 1 hour after resuscitation were detected by Western blotting and real-time RT-PCR. The encoding gene content, ELISA method to detect serum VAP-1 content and activity. The rats in the recovery group were treated with 2-bromoethylamine (20 mg / kg) and the control group (1 mL / kg saline). The blood pressure after resuscitation, the intestinal mucosal injury (Chiu’s score) and the intestinal mucosal epithelial cell apoptosis (TUNEL), and compared 24h survival rate in rats. Results The levels of VAP-1 protein, the mRNA level of VAP-1 and the content and activity of VAP-1 in serum in the severe hemorrhagic shock group were higher than those in sham operation group (all P <0.05) Decreased, but still higher than the sham operation group. Compared with the saline control group, the rats in the resuscitation of 20mg / kg 2-bromoethylamine at 1h and 24h after shock resuscitation had higher blood pressure (P = 0.010 and 0.039, respectively), and Chiu’s score and intestinal mucosa The apoptosis index of epithelial cells was lower than that of saline resuscitation control group (P = 0.022 and 0.002 respectively), and the 24h survival rate of shock rats was also higher than that of resuscitation control group (90% vs 60%). Conclusions Severe hemorrhagic shock can increase the expression and activity of VAP-1 in rats, while liquid resuscitation can reduce this increase. Suppression of its activity can improve the severity of severe hemorrhagic shock and hypotension after resuscitation and the damage and apoptosis of intestinal mucosa Death, improve 24h survival rate in rats.