两种肠系膜淋巴液处理方法的确立及探讨  

Establishment and Discussion the Two Methods of Processing Mesenteric Lymph

作　　者：张睿[1] 何桂珍[1] 王玉康[1] 

机构地区：[1]中国医学科学院北京协和医学院 北京协和医院肠外肠内营养科 协和转化医学中心,北京100730

出　　处：《现代检验医学杂志》2012年第1期24-27,共4页Journal of Modern Laboratory Medicine

基　　金：基金项目：北京自然科学基金资助项目（7102127）.

摘　　要：目的为研究大鼠肠道缺血再灌注损伤时肠系膜淋巴液在“肠-淋巴”途径中的作用，建立淋巴液的两种处理方法。方法SPF级SD健康雄性大鼠，体重280-320g，随机分为：肠道缺血再灌注引流（I／R＋D）组和普通引流（N＋D）组（n=10）。I／R＋D组夹闲肠系膜上动脉60min后复灌120min，同时引流肠系膜淋巴液180min；N＋D组开腹后只引汽淋巴液180min。收集肠系膜淋巴液。两组的淋巴液处理：①用蛋白酶K水解法降解其中蛋白质，并测定水解前后淋巴液中蛋白含量；②内毒素去除柱除去淋巴液中内毒素，并检测过柱前后淋巴液中内毒素水平。结果大鼠肠道缺血再灌注损伤后，淋巴液中的蛋白质含量及内毒素水平显著高于对照组（t值分别为-8．32和-9．91，P〈0．05差异有统计学显著性意义）。用蛋白酶K水解后能降解淋巴液中大部分蛋白质，处理前后蛋白含量：N＋D组（24．35±1．37）vs（2．94±0．33）g／L，I／R＋D组（34．89±1．68）g／Lvs（3．72±0．51）g／L，其水解效率都在87％以上；内毒素去除柱能除去淋巴液中大部分内毒素，处理前后淋巴液中内毒素水平：N＋D组（0．0095±0．0053）EU／ml vs（0．0021±0．0005）EU／ml，I／R＋D组（0．0321±0．0131）EU／ml vs（0．0030±0．0003）EU／ml．其去除效率I／R＋D组达到90％。结论肠道缺血再灌注损伤后淋巴液中蛋白质舍量和内毒素水平显著增高；蛋白酶支能够降解淋巴液中87％以上的蛋白质，内毒素去除柱能够通过亲和的方法去除淋巴液中大部分内毒素，为进一步研究“肠-淋巴”途径中淋巴液成分在肠道缺血再灌注损伤中的作用提供了新的方法及思路。Objective To research the role of the mesenteric lymph fluid in ＂gut-lymph pathway＂ during rat intestinal ischemia-reperfusion （I/R） injury,establish and discuss the methods of processing mesenteric lymph fluid. Methods 20 healthy male rats of SPF grade randomly was divided into 9, groups：intestinal iscbemia-reperfusion＋ drainage group （I/R＋D） and normal drainage （N-q-D） group. The rats in I/R＋D group were subjected to 60 min ischemia by clapping the superior mesenteric artery,followed by 120 min reperfusion. At the same time, the rats in I/R＋D group were treated with intestinal lymph drainage for 180 min. Rats in N＋D group were only treated with intestinal lymph drainage for 180 min. Collected the mesenteric lymph fluid of the two groups during the 180 min. The processing lymph fluid of two groups as followed：（1）Protei nase K＇degrades the protein of two groups, before and after that, detected the protein concentration. （2）Endotoxin removing columns bind and remove pyrogens from the lymph,detecting the endotoxin before and after the processing. Results Found that the levels of protein concentration and endotoxin of lymph fluid were significant higher than those of control group after intestinal （I/R） injury （ t value were - 8. 32 and - 9.91 separately, P〈 0. 05 was considered statistically significant）. Proteinase K could degrade the protein of two groups：N＋D （24. 35±1.37）g/L vs （2.94±0. 33）g/L,I/R＋D（34.89±1.68） g/L vs （3. 724±0.51） g/L,effieiency up to 87% ; Endotoxin removing columns could bind and removed pyrogens from the lymph：N＋D（0. 009 5±0. 005 3） EU/ml vs （0. 002 1±0. 000 5） EU/ml,I/R＋D（0. 032 1±0. 013 1） EU/ml vs （0. 003 0 ±0. 000 3） EU/ml,efficiency ups to 90% in I/R＋D group. Conclusion The levels of protein concentration and endotoxin of mesenteric lymph fluid were much higher than the normal control after intestinal （I/R） injury. Proteinase K and endotoxin removing columns could remove the most

关 键 词：肠道缺血再灌注 肠系膜淋巴液 肠淋巴液处理 淋巴液蛋白浓度 淋巴液内毒素 

分 类 号：R-332[医药卫生]