检验性失血对MODS大鼠血清促红细胞生成素、血红蛋白和器官功能的影响

Impacts of different amounts of laboratory-testing blood loss on the hemoglobin, erythropoietin and organ function of rats with septic multiple organ dysfunction syndrome

目的探讨不同检验性失血量对多器官功能障碍综合征(MODS)大鼠血红蛋白(Hb)、促红细胞生成素(EPO)、器官功能及肾脏炎症介质、氧化应激、细胞凋亡的影响。方法 120只SD大鼠盲肠结扎穿孔(CLP)术后按照随机数表法均分为四组:MODS组(M组)、MODS+检验性失血组1(ML1组)、MODS+检验性失血组2(ML2组)、MODS+检验性失血组3(ML3组),每组各30只。ML1组手术关腹后立即尾静脉切开放血1 ml后压迫止血,ML2组手术关腹后立即及术后24 h尾静脉切开各放血1 ml后压迫止血,ML3组手术关腹后立即、术后24 h及48 h尾静脉切开各放血1 ml后压迫止血。72 h统计各组病死率,每组取10只心脏取血2 ml,检测Hb、肌酸磷酸激酶同工酶(CK-MB)、谷丙转氨酶(ALT)、尿素氮(BUN)、肌酐(Cr)、EPO、肿瘤坏死因子(TNF-α),取肾脏组织检测核转录因子(NF-κB)、丙二醛(MDA)和末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口标记(TUNEL)阳性细胞数。结果制模72 h,M组死亡10只,病死率为33.3%,ML1组死亡13只,病死率为40%,ML2组死亡15只,病死率为50%,ML3组死亡18只,病死率为60%,两两比较差异具有统计学意义(P<0.05)。与M组比较,ML1、ML2、ML3组大鼠72 h Hb明显下降,CK-MB、ALT、BUN、Cr、TNF-α、肾脏NF-κB、MDA和TUNEL阳性细胞数明显增高,差异具有统计学意义(P<0.05)。ML1组肾脏EPO浓度比M组略有升高,但差异无统计学意义(P>0.05),ML2、ML3组肾脏EPO浓度比M组明显增高(P<0.05)。结论检验性失血可能通过贫血、上调EP0水平、炎症反应、氧化应激和促进细胞凋亡导致MODS大鼠器官功能障碍加重并影响预后。

Objective To discuss the impacts of different amounts of laboratory-testing blood loss on the he-moglobin （Hb）, erythropoietin （EPO）, organ function, renal inflammatory medium, oxidative stress and cell apoptosis of the rats with septic multiple organ dysfunction syndrome （MODS）. Methods One hundred and twenty SD rats were averaged into four groups through the random number table after CLP： MODS group （M）, MODS＋laborato-ry-testing blood loss group 1 （ML1）, MODS＋laboratory-testing blood loss group 2 （ML2）, MODS＋laboratory-testing blood loss group 3 （ML3）, with 30 rats in each group. When the surgery was finished and the abdomen was closed, 1 ml of blood was drawn immediately after the tail vein phlebotomy in group ML1, then compression was performed to stop bleeding. In group ML2, 1 ml of tail vein blood was taken respectively immediately after the surgery and 24 h af-ter surgery, while in group ML3, blood samples was collected 24 h after surgery and 48 h after surgery. Fatality rate was calculated 72 h after the surgery. Ten rats in each group was selected and 2 ml of blood was taken from the heart for detecting Hb, creatine kinase MB （CK-MB）, alanine aminotransferase （ALT）, urea nitrogen （BUN）, creatinine （Cr）, EPO, tumor necrosis factorα（TNF-α）. Malondialdehyde （MDA）, nuclear factor-κB （NF-κB） in the kidney tis-sue, as well as the amount of TUNEL positive cells were also detected. Results After 72 h of moulding, 10 rats died in the group M and the fatality rate was 33.3%, with 13 （40%） died in group ML1, 15 （50%） died in group ML2 and 18 （60%） died in group ML3. The differences between each two groups were statistically significant （P〈0.05）. Compared with group M, the Hb of group ML1, ML2 and ML3 all declined significantly, while the levels of CK-MB, ALT, BUN, Cr, TNF-α, MDA, NF-κB, and the amount of TUNEL positive cells all increased significantly （P〈0.05）. EPO concen-tration in group ML1 was slightly higher than that in group M, but with no statistical difference （P〉0.05）. The EPO con-centration in the group ML2 and ML3 was significantly higher than that in group M （P〈0.05）. Conclusion Laborato-ry-testing blood loss could aggravate the organ function disorder and affect the prognosis of rats with septic MODS by anemia, EPO level up-regulation, inflammatory reaction, oxidative stress and cell apoptosis promotion.