乌司他丁对烧冲复合伤大鼠急性肺损伤及凝血参数时相性变化的影响

Influences of ulinastatin on acute lung injury and time phase changes of coagulation parameters in rats with burn-blast combined injuries

目的探讨乌司他丁对重度烧冲复合伤大鼠急性肺损伤及凝血参数时相性变化的影响。方法将192只SD大鼠按随机数字表法分为单纯烧冲复合伤组、乌司他丁＋烧冲复合伤组、假伤组，每组64只。将2组烧冲复合伤大鼠采用自制新型爆炸装置致中度冲击伤后，将致伤大鼠背部浸人94℃热水中12s，造成25％体表总面积Ⅲ度烫伤（以下称烧伤）；假伤组大鼠仅背部浸入37℃温水中12s模拟致伤。3组大鼠伤后即刻腹腔注射乳酸林格液40mL／kg；乌司他丁＋烧冲复合伤组大鼠另腹腔注射乌司他丁4×10^4U／kg，每隔12小时注射1次，持续至伤后72h。伤前和伤后3、6、12、24、48、72h及7d，每组取8只大鼠取腹主动脉血检测血浆活化部分凝血活酶时间（APTT）、凝血酶原时间（PT）及纤维蛋白原、D-二聚体、抗凝血酶Ⅲ（AT-Ⅲ）、α2-抗纤溶酶（α2-AP）水平。伤后24h，各组取3只经取血检测凝血指标后的大鼠，处死后观察肺部大体损伤情况；伤后72h，同前取3只大鼠处死后行肺组织病理学观察。对数据行析因设计方差分析和LSD检验。结果（1）与假伤组比较，单纯烧冲复合伤组大鼠APTT伤后72h、7d明显延长（P〈0．05或P〈0．01），PT伤后3、72h明显延长而伤后6h明显缩短（P〈0．05或P〈0．01），纤维蛋白原水平伤后12h～7d明显升高（P〈0．01），AT—Ⅲ水平伤后6、12h明显降低（P〈0．01），仪2．AP水平伤后6h明显降低而伤后24～72h明显升高（P〈0．01）。与单纯烧冲复合伤组比较，乌司他丁＋烧冲复合伤组大鼠APTT伤后3、6h明显延长（P〈0．01），PT伤后3、12、72h明显缩短（P〈0．05或P〈0．01），纤维蛋白原水平伤后6、12h明显降低而伤后72h明显升高（P〈0．05或P〈0．01），AT-Ⅲ水平伤后3、12、48、72h明显升高（P〈0．05或P〈0．01）；α2-AP水平伤后12～72h明显降低（P〈0．05或P〈0．01）。假伤组、单纯烧冲复合伤组、乌司他丁＋烧冲复合伤组大鼠伤前和伤后3、6、12、24、48、72h及7dD-二聚体水平分别为（0．084±0．013）、（0．115±0．015）、（0．158±0．022）、（0．099±0．011）、（0．099±0．012）、（0．089±0．011）、（0．124±0．014）、（0．116±0．018）μg／mL，（0．064±0．033）、（0．114±0．016）、（0．135±0．009）、（0．060±0．008）、（0．104±0．010）、（0．124±0．020）、（0．180±0．036）、（0．201±0．032）μg／mL，（0．074±0．013）、（0．084±0．035）、（0．101±0．050）、（0．091±0．046）、（0．096±0．034）、（0．044±0．019）、（0．106±0．049）、（0．118±0．047）μg／mL。与假伤组比较，单纯烧冲复合伤组大鼠D-二聚体水平伤后6、12h明显降低而伤后48h～7d明显升高（P〈0．05或P〈0．01）；与单纯烧冲复合伤组比较，乌司他丁＋烧冲复合伤组大鼠D-二聚体水平伤后3、48、72h及7d明显降低（P〈0．05或P〈0．01）。（2）伤后24h，单纯烧冲复合伤组大鼠胸腔有大量淡红色积液，双侧肺叶均充血、水肿明显，左侧肺中、下叶有少量血凝块；乌司他丁＋烧冲复合伤组大鼠胸腔有少量淡黄色积液，双侧肺叶充血、水肿较单纯烧冲复合伤组明显减轻，左侧肺叶无血凝块；假伤组大鼠胸腔无积液，未见充血、水肿或出血。（3）伤后72h，单纯烧冲复合伤组大鼠肺泡结构紊乱，肺泡腔萎陷，肺间质水肿、增厚，有大量炎性细胞浸润，肺泡间隔明显断裂，并可见透明血栓形成；乌司他丁＋烧冲复合伤组大鼠肺泡结构较单纯烧冲复合伤组明显改善，肺泡腔萎陷明显减少，肺间质水肿有所减轻，炎性细胞浸润明显减少，肺泡间隔无断裂，无血栓形成；假伤组大鼠肺泡腔充盈良好，未见肺间质水肿或炎性细胞浸润，无血栓形成。结论乌司他丁对重度烧冲复合伤大鼠急性肺损伤有积极的治疗作用，对烧冲复合伤造成的凝血、纤溶功能紊乱具有很好的调节作用。

Objective To explore the influences of ulinastatin on acute lung injury and time phase changes of coagulation parameters in rats with severe burn-blast combined injuries. Methods One hundred and ninety-two Sprague-Dawley rats were divided into pure burn-blast combined injury group, ulinastatin ＋ burn-blast combined injury group, and sham injury group according to the random number table, with 64 rats in each group. Two groups of rats with combined burn-blast injuries were inflicted with moderate blast injuries with the newly self-made explosive device. Then the rats were inflicted with 25% total body surface area full-thickness scald （hereinafter referred to as burn） on the back by immersing in 94 ~C hot water for 12 s. Rats in sham injury group were sham injured on the back by immersing in 37 ℃ warm water for 12 s. Immediately after injury, rats in the three groups were intraperitoneally injected with Ringer＇s lactate solution （40 mL/kg） , meanwhile rats in ulinastatin ＋ burn-blast combined injury group were intraperitoneally injec- ted with ulinastatin （4 × 10^4 U/kg） , once every 12 hours, until post injury hour （PIH） 72. Before injury, at PIH 3, 6, 12, 24, 48, 72, and on post injury day （PID） 7, 8 rats in each group were selected to harvest abdominal aortic blood samples to detect plasma levels of activated partial thromboplastin time （APTT） , prothrombin time （PT）, fibrinogen, D-dimer, antithrombin Ⅲ（AT-Ⅲ）, and α2-antiplasmin （α2-AP）. At PIH 24, three rats in each group which were used in detection of coagulation parameters were sacrificed to observe lung injury. At PIH 72, three rats in each group were sacrificed for histopathological observation of lung. Data were processed with analysis of variance of factorial design and least-significant difference test. Results （1） Compared with those of rats in sham injury group, APTT of rats in pure burn-blast combined injury group significantly prolonged at PIH 72 and on PID 7 （ P 〈 0.05 or P 〈 0.01 ）. PT significantly prolonged at PIH 3 and 72 and significantly shortened at PIH 6 （ P 〈 0.05 or P 〈 0.01 ） . Fibrinogen level significantly increased from PIH 12 to PID 7 （ P 〈0.01 ）. AT-Ⅲ level significantly decreased at PIH 6 and 12 （ P 〈 0.01 ） , and α2-AP level significantly decreased at PIH 6 and significantly increased from PIH 24 to 72 （ P 〈 0.01 ）. Compared with those of rats in pure burn-blast combined injury group, APTT of rats in ulinastatin ＋ burn-blast combined injury group significantly prolonged at PIH 3 and 6 （ P 〈 0.01 ） while PT significantly shortened at PIH 3, 12, and 72 （ P 〈 0.05 or P 〈 0.01 ）. Fibrinogen level significantly decreased at PIH 6 and 12 and significantly increased at PIH 72 （ P 〈 0.05 or P 〈 0. 01 ）. AT-Ⅲ level significantly increased at PIH 3, 12, 48, and 72 （ P 〈 0.05 or P 〈 0.01 ） , and ct2-AP level significantly decreased from PIH 12 to 72 （ P 〈0.05 orP 〈0.01 ）. D-dimer level of rats in sham injury group, pure burnblast combined injury group, and ulinastatin ＋ burn-blast combined injury group were respectively （0. 084 ± 0.013）, （0.115±0.015）, （0.158±0.022）, （0.099±0.011）, （0.099±0.012）, （0.089±0.011）, （0.124±0.014）, and （0. 116 ±0.018）μg/mL, （0.064 ±0.033）, （0. 114 ±0.016）, （0. 135 ± 0.009）, （0.060±0.008）, （0. 104±0.010）, （0. 124±0.020）, （0. 180±0.036）, and （0.201 ±0.032） μg/mL, （0.074 ±0.013）, （0.084 ±0.035）, （0.101±0.050）, （0.091±0.046）, （0.096±0.034）, （0.044 ±0.019）, （0.106±0.049）, and （0.118 ±0.047） μg/mL. Compared with that of rats in sham injury group, D-dimer level significantly decreased at PIH 6 and 12 and significantly increased from PIH 48 to PID 7 （ P 〈0.05 orP 〈0.01）. Compared with that of rats in pure burn-blast combined injury group, D-dimer level of rats in ulinastatin ＋ burn-blast combined injury group significantly decreased at PIH 3, 48, and 72, and on PID7 （ P 〈0.05 orP 〈0.01）. （2） At PIH24, there was alarge amount of light red effusion in the thoracic cavity, and both lung lobes were hyperemic and edematous with a small amount of blood clots in the left and middle lobe of rats in pure burn-blast combined injury group. There was a small amount of yellowish effusion in the thoracic cavity of rats in ulinastatin ＋ burn-blast combined injury group, and the degree of hyperemic and edematous of bilateral lobes was lighter compared with rats in pure burn- blast combined injury group with no clot in the left lobe. No congestion, edema, or bleeding was observed in lungs of rats in sham injury group. （3） At PIH 72, disorganized alveolar structure, collapsed alveolar cavi- ty, edematous and thickening pulmonary interstitium, infiltration of a large amount of inflammatory cells, ob- vious rupture of alveolar septum, and hyaline thrombus were observed in lungs of rats in pure burn-blast combined injury group. Significantly improved alveolar structure, less collapsed alveolar cavity, improved edematous pulmonary interstitium, less infiltration of inflammatory cells, rupture of alveolar septum, and no thrombus were observed in lungs of rats in ulinastatin ＋ burn-blast combined injury group. The lung tissue had a well-filled alveolar cavity with no interstitial edema or infiltration of inflammatory cells and no thrombosis in lungs of rats in sham injury group. Conclusions Ulinastatin has positive therapeutic effects on acute lung injury in rats with severe burn-blast combined injuries through its good regulating effects on coagulation and fibrinolytic disorders caused by burn-blast combined injuries.