乌司他丁对重度脓毒症诱导的ALI/ARDS模型大鼠肺组织中蛋白C及活化蛋白C含量的影响

Effects of Ulinastatin on Concentrations of Protein C and Activated Protein C in Lung Tissues from Rats Model of Severe Sepsis-induced ALI/ARDS

目的：探讨乌司他丁对重度脓毒症诱导的急性肺损伤（acute lung injury,ALI）/急性呼吸窘迫征（acute respiratory distress syndrome,ARDS）模型大鼠肺组织中蛋白C（protein C,PC）及活化蛋白C（activated protein C,APC）含量的影响。方法：对27只体质量为200-250 g的清洁级雄性SD大鼠采用盲肠结扎穿孔法联合0.9%氯化钠溶液灌洗法制作重度脓毒症诱导的ALI/ARDS大鼠模型,以PaO2/FiO2≤300作为ALI/ARDS模型大鼠纳入标准,随机分为对照组、乌司他丁5万U/kg组、乌司他丁10万U/kg组,每组9只。各组均在造模成功后立即予乌司他丁或0.9%氯化钠溶液静脉注射,乌司他丁5万U/kg组予乌司他丁5万U/kg静脉注射,乌司他丁10万U/kg组予乌司他丁10万U/kg静脉注射,对照组予0.9%氯化钠溶液静脉注射,静脉注射持续时间为10 min。造模后1、2、3 h分批处死大鼠,取肺组织制备组织匀浆,采用酶联免疫吸附试验（ELISA）法测定其PC及APC含量。结果：（1）乌司他丁10万U/kg组大鼠造模后1、2、3 h肺组织PC的含量分别为（5196±981）ng/mL、（6156±1256）ng/mL、（6481±1141）ng/mL,均高于乌司他丁5万U/kg组[（3691±812）ng/mL、（4367±1547）ng/mL、（5313±1356）ng/mL]和对照组[（3676±579）ng/mL、（3248±851）ng/mL、（3188±1808）ng/mL],且差异有统计学意义（P〈0.01）;（2）乌司他丁10万U/kg组大鼠造模后1、2、3 h肺组织APC含量分别为（2798±380）ng/mL、（2964±731）ng/mL、（3156±805）ng/mL,均高于乌司他丁5万U/kg组[（1596±516）ng/mL、（1994±196）ng/mL、（1712±456）ng/mL],但差异无统计学意义（P〉0.05）;两组APC含量均高于对照组[（1459±395）ng/mL、（1300±348）ng/mL、（1141±318）ng/mL],差异均有统计学意义（P〈0.05）。各组大鼠PC和APC含量组内比较差异均无统计学意义（P〉0.05）。结论：乌司他丁有助于提高重度脓毒症诱导的ALI/ARDS模型大鼠肺组织PC和APC的含量,并呈剂量依赖趋势,它可能对脓毒症诱导的ALI/ARDS大鼠肺组织有保护作用。

Objective：To investigate the effects of ulinastatin on concentrations of protein C（PC）and activated protein C（APC）in lung tissues from rat model of severe sepsis-induced acute lung injury（ALI）/acute respiratory distress syndrome（ARDS）.Methods：Rat models with severe sepsis-induced ALI/ARDS were made by cecal ligation and puncture combined with normal saline lavage,taking PaO2/FiO2 ≤300 as inclusion criteria.A total of ALI/ARDS rats were randomly divided into control group ulinastatin 50 000 units/kg group and ulinastatin 100 000 units/kg group,with 9 rats in each group.Ulinastatin of 50 000 units/kg,ulinastatin of 10 000 units/kg and 0.9%sodium chloride solution were administered intravenously within 10 minutes in ulinastrtin5000 units/kg group,ulinastatin10 0000 units/kg group and control group,respectively,immediately after models were constructed.Rats were sacrificed at 1h,2h,3 h were constructed after models,and lung tissue homogenates were prepared after tissue weighing.Tissue concentrations of PC and APC were detected by enzyme-linked immunosorbent assay（ELISA）method.Results：（1）The PC concentrations of lung tissues in ulinastatin 100 000 units/kg group were（5196± 981）ng/mL,（6156±1256）ng/mL,（6481±1141）ng/mL,respectively,at 1 h,2 h,3 h after models were constructed,which were higher than those in ulinastatin50 000 units/kg group[（3691±812）ng/mL,（4367±1547）ng/mL,（5313±1356）ng/mL] and those in control group[（3676±579）ng/mL,（3248±851）ng/mL,（3188±1808）ng/mL];and there were significant differences（P0.01）.（2）The concentrations of APC in ulinastatin100 000 units/kg group were（2798±380）ng/mL,（2964±731） ng/mL,（3156±805）ng/mL,respectively,at 1 h,2 h,3 h after models were constructed,which were higher than those in ulinastatin 50 000 units/kg group[（1596±516）ng/mL,（1994±196）ng/mL,（1712±456）ng/mL],but there was no differences.The APC concentrations of the above two groups were higher than those in control group[（1459±395）ng/mL,（1300±348） ng/mL,（1141±318）ng/mL]（P0.05）.There were no significant differences in concentrations of PC and APC within each group at different time points.Conclusions：Ulinastatin may increase the concentrations of PC and APC in rat lung tissues and it showed a dose-dependent trend.Moreover,ulinastatin plays a protective role in rat lung with sepsis-induced ALI/ARDS.