乌司他丁对幼猪体外循环心脏手术脑损伤影响的实验研究

Experimental study of the effect of ulinastatin on brain injury caused by deep hypothermic low-flow cardiopulmonary bypass surgery in piglets

目的评价广谱蛋白酶抑制剂乌司他丁对幼猪深低温低流量体外循环（cardiopulmonary bypass，CPB）心脏手术脑损伤的影响。方法普通幼猪15只，采用随机数字表法分为3组（每组5只）：乌司他丁组（U组）、对照组（c组）和假手术组（S组）。U组和C组麻醉后正中开胸，常规建立CPB，降温至25℃后，低流量循环（50ml·kg-1．min。），主动脉阻断120rain后开放，调整停机后观察120min，取海马组织。U组在CPB开始和主动脉开放即刻分别给予乌司他丁1．2万IU／kg，C组给予等量生理盐水。S组麻醉、单纯开胸，即取脑海马组织。3组在麻醉诱导后即刻（TI）、U组和c组在停机后5min（T2）、120min（T，）取静脉血，ELISA法测神经元特异性烯醇化酶（neuron-specificenolase，NSE）、S100蛋白B亚型（S100proteinβsubtypes，S100β）、髓鞘碱性蛋白（myelinbasicprotein，MBP）。光镜下观察海马病理学结果，测含水量。结果15只幼猪均完成至实验终点。与T1相比，C组在T2、T3时点NSE、S10013、MBP分别升高30．2、32．3、62．0％和33．1、29．8、57．1％，差异均有统计学意义（P〈0．05）；U组在T2、T3时点NSE、S10013、MBP分别低于相应时点C组16．8、7．3、20．4％和20．4、2．7、18．5％，差异均有统计学意义（P〈O．05）。C组、U组均可见脑损伤，U组较轻。含水量C组比U组升高2．2％（P〈0．05）。结论深低温低流量CPB心脏手术对幼猪存在脑损伤，乌司他丁具有对抗脑损伤（脑保护）作用。

Objective To evaluate the effect of ulinastatin on brain injury caused by deep hypothermic low-flow cardiopulmonary bypass （CPB） cardiac surgery in piglets. Methods 15 general-type piglets were assigned into ulinastatin group （group U, n=5）, control group （group C, n=5） and sham operation group （group S, n=5）, randomly. Routine CPB was established after median thoracotomy in group U and C under anesthesia. When the temperature of piglets dropped to 25 ℃, low-flow CPB （50 ml·kg-1·min-1） began. After 120min of aortic cross-clamping, the aortic was declamped and finally weaned from CPB and piglets were sacrificed and brain hippocampus tissue was harvested following 120 min observation after CPB ceasation, 12 000 IU/kg of ulinastatin was given instantly when CPB began and aortic declamp in group U respectively, and group C received equivale saline. Brain hippocampus tissue was harvested after anesthesia and median thoracotomy in group S. Venous blood samples were taken immediately after anesthesia induction （T1） in all groups, 5 min（T2） and 120 min（T3） after the CPB shutdown in both Group U and C, for the test of neuron-specific enolase （NSE）, S100 protein 13 subtypes （S10013） and myelinbasicprotein （MBP） by ELISA method. Pathology results of hippocampus were observed by light microscopy, and their water content were also measured. Results All 15 piglets completed experiment end. In group C, compared with Tl timepoint, NSE, S100J3, MBP were significantly increased by 30.2, 32.3, 62.0% in T2 and 33.1, 29.8, 57.1% in T3 timepoints respectively （P〈0.05）. Compared with group C,NSE, S10013, MBP were significantly decreased by 16.8, 7.3, 20.4% in T2 and 20.4, 2.7, 18.5%, in T3 timepoints respectively in group U （P〈0.05）. Brain injury was observed in both group U and C, and was lighter in group U. Water content of hippocampus was increased by 2.2% in group C than that of group U （P〈O.05）. Conclusions Deep hypothermic low-flow cardiopulmonary bypass surgery would result in brain injury on piglets, and ulinastatin could inhibit against those injury.