乌司他丁不同处理方式对体外循环患者血管内皮细胞的保护作用

Protective Effects of Differert Conditionings of Ulinastatin on Vascular Endothelial Cells during Cardiopulmonary Bypass

目的探讨乌司他丁对体外循环（CPB）患者血管内皮细胞保护作用及机制。方法选择择期在体外循环下行风湿性心脏病瓣膜置换术患者80例,随机分为A组（空白对照组）、B组（预处理组）、C组（后处理组、D组（联合处理组）,每组20例;B组予乌司他丁2万U/kg,于麻醉后,升主动脉阻断前10 min自中心静脉输入;C组予乌司他丁2万U/kg,于升主动脉阻断后加入CPB预充液中;D组先以乌司他丁1万U/kg于升主动脉阻断前10 min自中心静脉输入,后以乌司他丁1万U/kg于升主动脉阻断后加入预充液中。分别于CPB前（T_1）、升主动脉开放5 min（T_2）、CPB结束后30 min（T_3）、CPB结束后4 h（T_4）时采血测定血浆肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6水平及超氧化物歧化酶（SOD）和丙二醛（MDA）浓度。采用标记特异性抗体CD146的免疫磁珠技术分离循环内皮细胞（CEC）并计数,采用酶联免疫吸附测定（ELISA）法检测血浆血栓调节蛋白（TM）和血管性假血友病因子（vWF）水平。结果与A组比较,B,C,D组TNF-α浓度于T_2～T_3,IL-6浓度于T_2～T_4均明显降低,差异有统计学意义（P〈0.05）;与A组比较,各组SOD血浆浓度在T_2～T_4明显升高,各组MDA血浆浓度在T_2～T_3明显降低,差异有统计学意义（P〈0.05）;B,C,D组TM浓度在T_2～T_3,B组T_4较A组明显降低;B组vWF浓度在T_2及T_4,C组在T_2～T_3,D组在T_2～T_4较A组明显降低,差异有统计学意义（P〈0.05）;与A组比较,B,C,D组CECs数目于T_3点明显减少,差异有统计学意义（P〈0.05）。结论乌司他丁预处理和后处理均对体外循环患者血管内皮细胞具有一定的保护作用,其可能通过抑制炎症因子和氧自由基发挥作用。

Objective To investigate the protective effect and mechanisms of ulinastatin on vascular endothelial cells during cardiopul-monary bypass. Methods Eighty patients undergoing mitral valve and aortic valve replacement under CPB were randomly divided into 4 groups, 20 cases in each group： Group A（ control）; Group B（ pre- conditioning）, Ulinastatin 20 000 U/kg was injected to central vein after anesthesia and the aorta block before 10 minutes; Group C（ post- conditioning）, Ulinastatin 20 000 U/kg was added to CPB pre liquid after ascending aorta block; Group D（ co- conditioning）, Ulinastatin 10 000 U/kg was injected to central vein before ascending aorta block firstly, then the left was added to CPB pre liquid after ascending aorta block. Blood samples were taken before CPB（ T1baseline）, 5 min after aortic unclamping（ T2）, 30 min（ T3） and 4h（ T4） after termination of CPB for determination of plasma concentrations of TNF- α, IL- 6, SOD and MDA. CECs were isolated and counted by using dynabeads coated with the specific antibody of CD146. ELISA was used to detect the levels of plasma TM and vWF. Results Compared with group A, the concentration of TNF- α at T2- 3 and IL- 6 at T2- 4 were significantly lower in group B, C and D and the plasma concentration of SOD was significantly increased at T2- 4 and MDA was significantly lower at T2- 3 in group B, C and D. Compared to group A, the plasma TM concentration were significantly lower at T2- 3 in group B, C, D and at T4 in group B than in group A. The plasma vWF concentration were significantly lower at T2, 4 in group B, T2 ~ 3 in group C, T2- 4 in group D as compared with group A. The number of CECs at T3 in group B, C and D was significantly decreased than that in group A. Conclusion Ulinastatin pre- conditioning and post- conditioning has certain protective effects on vascular endothelial cells during cardiopulmonary bypass in patients undergoing open heart surgery,which may relies on inhibiting the inflammatory factors and oxygen free radicals.