无肝素化体外循环转流及早期炎性因子的表达

Heparin-free Cardiopulmonary Bypass and Early Systemic Inflammatory Response

目的探讨采用鱼精蛋白-琼脂糖凝胶进行无肝素化体外循环(CPB)的可行性。方法选择健康成年家犬12只,犬龄2～3岁,雌雄不限,体重20～28(23.3±3.7)kg。按随机数字表法将12只犬分为两组,每组6只,肝素化组:行常规CPB;非肝素化组:采用鱼精蛋白-琼脂糖凝胶吸附血浆凝血因子,即行无肝素化CPB。于CPB开始时和CPB1 h、2 h、3 h采用酶联免疫吸附法(ELISA)测定动脉血中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素8(IL-8)的浓度,并进行比较。结果非肝素化组和肝素化组犬在CPB过程中肉眼观察膜肺内均未见血栓形成,全血激活凝血时间(ACT)始终>480 s,生命体征均平稳。两组血浆TNF-α、IL-6和IL-8浓度在CPB开始时差异无统计学意义,非肝素化组血浆TNF-α和IL-8于CPB 1 h、CPB 2 h和CPB 3 h时均高于肝素化组[CPB3 h TNF-α:(156.48±16.65)ng/L vs.(115.87±15.63)ng/L,t=4.356,P=0.001;CPB 3 h IL-8:(365.38±46.18)ng/L vs.(299.29±34.50)ng/L,t=2.808,P=0.019],而两组IL-6浓度差异无统计学意义(P>0.05)。结论依靠鱼精蛋白-琼脂糖凝胶对凝血因子进行吸附,以进行无肝素化CPB效果明确,但具有一定的促进全身炎症反应效应。

Objective To explore the feasibility of using protamine-agarose gel to achieve heparin-free cardiopul- monary bypass （CPB）. Methods A total of 12 healthy adult dogs were chosen, the dogs were between 2-3 years old, either male or female, with their mean body weight of 23.3 ± 3.7 kg （ranging from 20 to 28 kg ）. All the dogs were randomly divided into two groups with 6 dogs in each group. In the heparinized group, conventional CPB technique was used; in the non-heparinized group, protamine-agarose gel column was used to absorb plasma clotting factors in CPB without use of heparin. At the beginning of CPB and 1 h, 2 h, 3 h after CPB, arterial blood samples were collected from dogs in both groups. The expression levels of tumor necrosis factor-a （TNF-a）, interleukin-6 （IL-6） and interleukin-8 （IL-8） were measured by enzyme-linked immunosorbent assay （ELISA）and compared. Results There was no thrombus formation in the membrane oxygenators during CPB by naked eye observation in both groups. Activated coagulation time （ACT） was always greater than 480 s during CPB. The vital signs of the dogs were all stable during CPB. At the beginning of CPB, there was no statistical difference in plasma concentrations of TNF-a, IL-6, IL-8 between the two groups. At 1 h, 2 h and 3 h after CPB, the expression levels of TNF-a and IL-8 of the non-heparinized group were significantly higher than those of the heparinized group （CPB 3 h TNF-a： 156.48 ± 16. 65 ng/L vs. 115.87 ± 15.63 ng/L, t= 4. 356, P=-0.001 ; CPB 3 h IL-8： 365.38±46. 18 ng/L vs. 299. 29±34. 50 ng/L, t=2. 808, P=0. 019）. There was no statistical difference in the expression level of IL-6 between the two groups （P 〉 0.05）. Conclusion Using protamine-agarose gel to absorb plasma clotting factors is an effective technique to establish heparin-free CPB. But this method can induce significant systemic inflammatory response.