大鼠肠淋巴液引流方法的建立及肠道缺血再灌注损伤时活性物质的改变

Method for Drainage of Lymph Fluid and Determining the Change of Active Materials in Lymph Fluid after Rat Ischemia-reperfusion Injury

目的建立大鼠肠淋巴液引流方法并探讨可能参与肠道缺血再灌注损伤的活性物质。方法建立并熟练掌握大鼠肠淋巴液引流的方法。选用SPF级SD大鼠,随机分为肠道缺血再灌注加淋巴液引流组(I/R+引流组)和单纯肠淋巴液引流组(引流组),其中I/R+引流组大鼠行肠系膜上动脉夹闭,缺血60min,再灌注120min。收集大鼠淋巴液和血清,比较两组内毒素、高迁移率族蛋白1(HMGB1)及细胞因子,包括肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和可溶性细胞黏附分子(sICAM-1)的水平。结果大鼠肠淋巴液引流的成功率较高(84.2%)。I/R+引流组的淋巴液和血清中内毒素、HMGB1以及炎症因子TNF-α、IL-1β、IL-6和sICAM-1均显著高于单纯引流组(P<0.05)。结论建立起比较简便、易行的肠淋巴液引流方法。内毒素、HMGB1和部分炎症因子可能参与了肠道缺血再灌注损伤。

Objective To set up a method for the drainage of active materials in lymph fluid and serum after rat ischemia-reperfusion lymph fluid and explore the change of injury. Methods The method of the drainage of lymph fluid was well established. Sixteen healthy male rats of SPF grade were selected and randomly divided into 2 groups ： intestinal ischemia-reperfusion ＋ drainage group （ I/R ＋ drainage group） and drainage group. All the rats were subjected to superior mesenteric artery occlusion for 60 minutes, followed by 120 minutes of reperfusion. We compared the change of high mobility group box-1 （ HMGB1 ） protein, endotoxin, tumor necrosis factor α （TNF-α）, interleukin （IL） -113, IL-6, and soluble vascular cell adhesion molecular- 1 （ slCAM-1 ） by draining lymph fluid and collecting serum in 2 groups. Results The drainage of lymph fluid was successfully performed. The HMGBI, endotoxin, and cytokines in serum and lymph fluid were significantly higher in ischemia-reperfusion group than in drainage group （ P 〈 0.05 ）. Conclusions The method for drainage of lymph fluid is simple and feasible. Endotoxin, HMGB1, and some cytokines in serum and lymph fluid may mediate the ischemia-reperfusion injury.