内皮细胞生物反应器在猪脓毒血症中的应用

Role of endothefial bioreactor device in sepsis porcine model

目的观察内皮细胞生物反应器对猪脓毒血症的治疗作用。方法健康杂交家猪静滴内毒素（LPS，0．25mg·kg^-1）建立脓毒症模型，随机（随机数字法）接受内皮细胞生物反应器（EBR组）和假性循环（对照组）治疗（每组n=6）。在LPS注射后立即进行体外循环，血流量为30mL／min，EBR组的血流经过内皮细胞生物反应器内腔后由颈内静脉回流至体内；假性循环组的血流经过同样大小的生物反应器，但不含有内皮细胞。同时记录每一头猪的生存时间。在实验开始前和开始后的每个小时分别观察实验动物的血流动力学指标、血液生化、血浆炎症因子、内皮素-1（ET-1）和血管性血友病因子（vWF）等。在记录实验猪的生存时间后立即取肺组织标本进行肺脏病理检查和肺损伤分数的计算。采用SPSS11．5统计软件对时间依赖的血流动力学和细胞因子数据进行重复测量方差分析，生存时间则使用Student’s t—tests进行检验。结果在注射LPS后两组的平均动脉压（MAP）都开始显著下降，但EBR组2h后MAP下降速度显著慢于对照组，两组差异具有统计学意义（P〈0．05）。EBR组的ET-1在治疗开始时有轻度下降，而对照组的ET-1则持续地升高，两组之间的差异具有统计学意义（P〈0．01），vWF在两组的变化都是先升高，并在实验6h时回复到基础水平，但EBR组的升高幅度低于对照组（P〈0．05）。肺损伤分数在EBR组显著低于对照组[（6．1±0．9）VS．（8．2±1．0），P〈0．05]。实验结束时EBR组和对照组的生存时间分别为[（6．7±1．32）hVS．（5．2±0．61）h，P〈0．01]。结论内皮细胞生物反应器不仅能够改善猪脓毒血症的血流动力学，而且能够维持重要脏器的功能，使得实验动物的存活时间延长。

Objective To investigate the role of endothelial bioreactor device in sepsis porcine model.Method Sepsis porcine model was induced by established endotoxin （LPS, 0.25 mg·kg^-1） in healthy hybrid swines. The animals were randomly divided （random number） into endothelial bioreactor device group （EBR group） and sham circulation group （Sham group） （ n = 6, respectively）. After the infusion of endotoxin, extracorporeal circulatio n was started with the blood flow of 30 mL/min. The blood went through the endothelial bioreactor, then went back to the body via internal jugular vein in the EBR group. The bioreactor with the same size and without endothelial cells （ECs） was used in the sham group. Hemodytkamie variables, blood biochemistry, inflammatory markers, Endothelin-1 （ET-1 ） and von Willebrand Factor （vWF） were examined just before and every hour after the injection. When the survival time of the animals was recorded, the animals were sacrificed to calculate the lung injury score. The time-dependent hemodynamies and cytokine data were compared between groups by repeated measurement ANOVA .Student＇s t -test was used to analyze the survival time. Results The mean arterial blood pressure （MAP） remarkably decreased in beth groups after LPS injection, while the decreasing rate in EBR group was significantly lower than that in control group after 2 hours （P 〈 0.05）. The ET-1 level in EBR group increased after a slight decrease at the beginning, while that in the sham group went on increasing （P 〈 0.01 ）. The vWF levels increased first, then returned to the baseline in the sixth hour in beth groups, while the change in EBR group was significantly less than that in the sham group （ P 〈 0.05）. The Lung Injury Score in EBR-treated group was significantly lower than that in the sham group （6.1 ± 0.9 vs. 8.2 ± 1.0, P 〈 0.05）. These physiologic and biochemical alterations were associated with a significant advantage to the survivals in the EBR group when compared with the control sham group （6.7 ± 1.32 vs. 5.2 ± 0.61 h, P 〈 0.01）. Conclusions Timely intervention in endotoxin shock with EC therapy by using tissue-engineered bioreactor may improve cardiovascalar performance and alter the natural course of this disease process, probably via modulating inflammation and coagulation cascades.