急性呼吸窘迫综合征家兔输注不同类型液体对凝血/纤溶功能的影响

Effects of infusion with different kinds of liquid on blood coagulation and fibrinolysis in rabbits with acute respiratory distress syndrome

目的 观察生理盐水、乳酸钠林格液及羟乙基淀粉130/0.4氯化钠注射液对油酸及内毒素二次打击诱导急性呼吸窘迫综合征（ARDS）家兔凝血/纤溶的影响.方法 将40只成年健康雄性家兔按随机数字表法分为假手术组、模型组、盐水组、林格组、胶体组,每组8只.经耳缘静脉序贯注射油酸0.1 mL/kg及大肠杆菌脂多糖（LPS）500μg/kg复制家兔ARDS模型.盐水组、林格组及胶体组于注射LPS后立即以7 mL·kg^-1·h^-1速度分别输注生理盐水、乳酸钠林格注射液、羟乙基淀粉130/0.4氯化钠注射液共210 min;假手术组及模型组不予输注液体.于注射LPS后30 min及210 min取动脉血测定血氧分压（PaO2）并计算氧合指数;于注射LPS后5、30、120、210 min取静脉血检测活化部分凝血活酶时间（APTT）、凝血酶原时间（PT）、纤维蛋白原（Fib）、抗凝血酶Ⅲ（AT-Ⅲ）、血清Ⅲ型前胶原肽（PⅢP）、组织型纤溶酶原激活物（t-PA）.实验结束后放血处死家兔,取肺组织进行免疫组化染色测定I型及Ⅲ胶原蛋白含量;测定肺组织湿/干质量（W/D）比值并进行肺组织病理评分.结果 与假手术组比较,模型组氧合指数于30 min和210 min显著降低;肺W/D比值及肺组织病理学评分升高;模型组APTT于30 min、PT于120 min开始逐渐延长;Fib进行性降低;AT-Ⅲ呈轻度降低趋势,各时间点均低于假手术组（均P〈0.05）;各时间点t-PA及PⅢP均升高,Ⅰ型及Ⅲ型胶原蛋白表达均显著增强.各输液组中,盐水组氧合指数、肺W/D比值及肺组织病理评分改善最明显,林格组无明显改变,而胶体组改善最差.盐水组APTT除120 min长于模型组外,30 min及210 min明显短于模型组（s：30 min：52.26±18.65比76.22±16.64,120 min：90.60±10.66比83.01±15.88,210 min：70.44±17.80比77.04±13.32,均P〈0.05）;林格组与胶体组APTT延长幅度均大于模型组,尤其以胶体组延长幅度最大;各输液组PT逐渐延长,于120 min及210 min均长于模型组（均P〈0.05）.各输液组Fib呈逐渐下降趋势,但盐水组下降幅度最小,而胶体组下降幅度最大;各输液组AT-Ⅲ与模型组存在类似下降趋势,且胶体组下降幅度最明显.林格组、胶体组及盐水组各时点t-PA均低于模型组（均P〈0.05）.林格组、盐水组各时间点血清PⅢP均显著低于模型组（μg/L：林格组：5min：250.60±36.53比285.77±65.55,30 min：248.73±44.41比302.16±37.73,120 min：249.14±43.16比296.09±38.64,210 min：246.62±44.72比295.45±42.75;盐水组：5 min：261.89±50.74比285.77±65.55, 30 min：247.71±50.40比302.16±37.73,120 min：246.58±42.27比296.09±38.64,210 min：222.73±18.51比295.45±42.75,均P〈0.05）,胶体组与模型组比较差异无统计学意义（P〉0.05）.盐水组、林格组及胶体组Ⅰ型及Ⅲ型胶原蛋白表达均低于模型组（P〈0.05或P〈0.01）,但胶体组Ⅰ型及Ⅲ型胶原蛋白高于林格组及盐水组（均P〈0.05）.结论 生理盐水、乳酸钠林格液及羟乙基淀粉130/0.4氯化钠对ARDS家兔凝血功能存在不同影响,以生理盐水影响最小,而羟乙基淀粉130/0.4氯化钠影响最大;3种液体均能减少ARDS家兔肺纤维组织成分,同时能不同程度减轻肺组织病理损害,其中生理盐水及乳酸钠林格液的作用程度大于羟乙基淀粉130/0.4氯化钠.

Objective To observe the influences of infusion with normal saline （NS）, Ringer solution and hydroxyethyl starch 130/0.4 sodium chloride on blood coagulation/fibrinolysis in rabbits with acute respiratory distress syndrome （ARDS） induced by two-hit of oleic acid （OA） and lipopolysaccharide （LPS）.Methods According to random number table, 40 healthy adult male rabbits were divided into sham operation, model, NS, Ringer and colloid groups （8 rabbits in each group）. The ARDS model was replicated by sequential injection of OA （0.1 mL/kg） and LPS （500μg/kg） into the ear marginal vein of rabbit. Immediately after injection of LPS, the NS, Ringer and colloid groups were treated by intravenous infusion of NS, lactate Ringer solution and hydroxyethyl starch 130/0.4 sodium chloride, respectively at a speed of 7 mL·kg^-1·h^-1 for 210 minutes. There was no liquid infusion in model and sham operation groups. At 30 minutes and 210 minutes after LPS injection, the arterial blood was collected and the partial pressure of arterial blood oxygen （PaO2） was measured and the oxygenation index （PaO2/FiO2） was calculated. At 5, 30, 120 and 210 minutes after LPS injection, venous blood was collected, and activated partial thromboplastin time （APTT）, prothrombin time （PT）, fibrinogen （Fib）, antithrombase Ⅲ （AT-Ⅲ）, serum procollagen peptide Ⅲ （PⅢP）, tissue plasminogen activator （t-PA） were measured, respectively. After the rabbits were killed by bloodletting at the end of experiment, the lung tissues were obtained, collagen Ⅰ and collagen Ⅲ in lung tissues were detected by immunohistochemistry staining, lung wet/dry weight ratio （W/D） and pathologic score of lung tissues were calculated.Results Compared with sham operation group, at 30 minutes and 210 minutes in model group the levels of PaO2/FiO2 were significantly decreased, and the lung W/D ratios as well as pathologic scores of pulmonary tissues were increased. In model group, the APTT began from 30 minutes while the PT began from 120 minutes to gradually prolong, and the value of Fib was progressively decreased; with a tendency of mild decline, the levels of AT-Ⅲ at all time-points were lower in model group than those in sham operation group （allP 〈 0.05）. The levels of t-PA and PⅢP at all time-points were significantly higher, and the expression levels of collagen Ⅰ and collagen Ⅲ in model group were obviously more strengthened compared to those in sham operation group. Among the three infusion groups, the improvement degrees of PaO2/FiO2, lung W/D ratio and pathologic score of pulmonary tissues were the highest in NS group, lowest in colloid group, and no significant changes in Ringer group. APTT in NS group except 120 minutes was longer, the APTTs at 30 minutes and 210 minutes were shorter in NS group than those in model group （s： 30 minutes： 52.26±18.65 vs. 76.22±16.64, 120 minutes： 90.60±10.66 vs. 83.01±15.88, 210 minutes： 70.44±17.80 vs. 77.04±13.32, allP 〈 0.05）; the prolongation of amplitudes of APTT in Ringer and colloid groups were greater than that in model group, particularly in colloid group, the greatest; the PT in three infusion groups were gradually prolonged, and at 120 minutes and 210 minutes were all longer than that in model group （allP 〈 0.05）. The levels of Fib in those treatment groups were all gradually decreased, the amplitude descent of Fib in NS group was the smallest and that in colloid group, the biggest; the levels of AT-Ⅲ in three infusion groups and model group had similar decline tendency, the descending amplitude being the most significant in colloid group. The levels of t-PA at all time-points in the three treatment groups were lower than those in model group （allP 〈 0.05）. The levels of PⅢP in serum at all time-points were lower in Ringer and NS groups than those in model group （μg/L： Ringer group： 5 minutes： 250.60±36.53 vs. 285.77±65.55, 30 minutes： 248.73±44.41 vs. 302.16±37.73, 120 minutes： 249.14±43.16 vs. 296.09±38.64, 210 minutes： 246.62±44.72 vs. 295.45±42.75; NS group： 5 minutes： 261.89±50.74 vs. 285.77±65.55, 30 minutes： 247.71±50.40 vs. 302.16±37.73, 120 minutes： 246.58±42.27 vs. 296.09±38.64, 210 minutes： 222.73±18.51 vs. 295.45±42.75, allP 〈 0.05）, but there were no statistically significant differences between the colloid group and model group. The expression levels of collagen Ⅰ and collagen Ⅲ in all liquid infusion groups were lower than those in model group （P 〈 0.05 orP 〈 0.01）, whereas in colloid group were higher than those in NS and Ringer groups （allP 〈 0.05）.Conclusions The infusion of NS, lactate Ringer solution and hydroxyethyl starch 130/0.4 sodium chloride have different influences on the blood coagulation function in ARDS rabbits, among which the effect of NS is the least, while of the hydroxyethyl starch 130/0.4 sodium chloride appears the greatest. The infusion of these three liquids can all decrease the pulmonary fibrous tissue in rabbits with ARDS, and in the mean time can alleviate the lung tissue pathological lesion for a certain degree, the effect of NS and Ringer solution being greater than that of hydroxyethyl starch 130/0.4 sodium chloride.