蛋白营养不良与能量营养不良状态下脓毒症大鼠凝血的变化  

Changes in coagulation of sepsis rats with protein-malnutrition or energy-malnutrition

作　　者：李栋 邹梦娟 王亮[1] 李鑫[1] 马晓春[1] Li Dong;Zou Mengjuan;Wang Liang;Li Xin;Ma Xiaochun(Department of Critical Care Medicine,the First Affiliated Hospital of China Medical University,Shenyang 110001,Liaoning,China)

机构地区：[1]中国医科大学附属第一医院重症医学科,沈阳110001

出　　处：《中华危重病急救医学》2019年第9期1113-1117,共5页Chinese Critical Care Medicine

基　　金：辽宁省重点临床科室诊疗能力建设项目(LNCCC-B01-2014)。

摘　　要：目的探讨蛋白营养不良和能量营养不良状态下脓毒症大鼠凝血的变化。方法将108只雄性SD大鼠按随机数字表法分为3组,每组36只。正常喂养组给予含18%蛋白饲料27 g/d自由饮食;蛋白营养不良组给予含5%蛋白饲料27 g/d低蛋白自由饮食;能量营养不良组给予含18%蛋白饲料9 g/d低能量饮食。连续喂养4周后,从各组分别选取8只大鼠进行营养不良评价,称量体重以及胸腺、脾脏重量,应用流式细胞仪检测脾脏T淋巴细胞亚群比例和M1型巨噬细胞比例,采用酶联免疫吸附试验(ELISA)检测血浆白细胞介素(IL-6、IL-10)水平。各组剩余28只大鼠用盲肠结扎穿孔术(CLP)制备脓毒症模型,其中20只用于Kaplan-Meier生存分析;其余8只于术后8 h处死,采用ELISA法检测血浆IL-6、IL-10水平,应用流式细胞仪检测脾脏M1型巨噬细胞比例,采用反转录-聚合酶链反应(RT-PCR)检测肝脏组织因子(TF)及凝血酶原激活物抑制因子-1(PAI-1)的mRNA表达。采用Pearson相关法对CLP术后大鼠TF、PAI-1的mRNA表达与IL-6进行相关性分析。结果①4周喂养结束后,正常喂养组和蛋白营养不良组大鼠体重增加,而能量营养不良组丢失初始体重的25%;蛋白营养不良组及能量营养不良组大鼠体重、胸腺和脾脏重量均明显低于正常喂养组。与正常喂养组和蛋白营养不良组比较,能量营养不良组大鼠脾脏CD3+T淋巴细胞、CD4+T淋巴细胞、M1型巨噬细胞比例以及血浆IL-6水平均明显升高〔CD3+T淋巴细胞比例:(52.1±3.7)%比(46.9±3.9)%、(44.5±2.2)%,CD4+T淋巴细胞比例:(35.0±3.6)%比(26.3±2.2)%、(25.1±2.3)%,M1型巨噬细胞比例:(8.7±2.0)%比(3.2±1.3)%、(4.2±1.1)%,IL-6(ng/L):129.4±16.2比48.1±10.0、53.0±8.3,均P<0.05〕。②CLP术后7 d Kaplan-Meier生存分析显示,能量营养不良组大鼠全部死亡,7 d累积生存率明显低于正常喂养组和蛋白营养不良组〔0%(0/20)比35%(7/20)、55%(11/20),均P<0.05〕。正常喂养组大Objective To investigate the changes in coagulation of sepsis rats with protein-malnutrition or energy-malnutrition.Methods 108 male Sprague-Dawley(SD)rats were divided into three groups by random number table,with 36 rats in each group.The rats in normal feeding group were given a free diet(27 g/d,containing 18%protein fodder),and the rats in protein-malnutrition group were given a low protein diet(27 g/d,containing 5%protein fodder).The rats in energy-malnutrition group were given a low energy diet(9 g/d,containing 18%protein fodder).After 4 weeks of continuous feeding,8 rats from each group were sacrificed for malnutrition evaluation.The weights of body,thymus and spleen were measured.The percentages of spleen T lymphocyte subsets and M1 macrophage were determined by flow cytometry.Plasma interleukins(IL-6 and IL-10)levels were determined by enzyme-linked immunosorbent assay(ELISA).The remaining 28 rats in each group were collected for cecal ligation and puncture(CLP)to reproduce the sepsis model,20 rats of which were used for Kaplan-Meier survival analysis,and the other 8 rats were sacrificed at 8 hours after CLP.The levels of plasma IL-6 and IL-10 were determined by ELISA,and the percentage of spleen M1 macrophages was determined by flow cytometry.The mRNA expressions of tissue factor(TF)and plasminogen activation inhibitor-1(PAI-1)in liver tissue were determined by reverse transcription-polymerase chain reaction(RT-PCR).Pearson correlation method was used to analyze the correlation between the mRNA expressions of TF and PAI-1 and IL-6 in rats after CLP.Results①After 4 weeks of feeding,the rats in the normal feeding group and protein-malnutrition group gained weight,while those in the energy-malnutrition group lost 25%of their initial body weight.The weights of body,thymus and spleen in the protein-malnutrition group and the energy-malnutrition group were significantly lower than those in the normal feeding group.Compared with the normal feeding group and the protein-malnutrition group,the percentages of s

关 键 词：营养不良 脓毒症 炎症反应 凝血活化 

分 类 号：R73[医药卫生—肿瘤]