大鼠肠系膜淋巴液持续引流新方法的初步建立

A Preliminary Method for Continuous Drainage of Mesenteric Lymph Fluid in Rats

目的通过建立大鼠肠系膜淋巴管-颈静脉辅助回流方法,实现肠系膜淋巴液的持续引流。方法将16只雄性SD大鼠随机均分为2组。对照组大鼠进行十二指肠和肠系膜淋巴管双插管手术,术后收集肠系膜淋巴液。实验组大鼠进行颈静脉和肠系膜淋巴管双插管手术,建立肠系膜淋巴管-颈静脉循环模型。术后第7天利用清醒活动装置收集肠系膜淋巴液,记录各组大鼠的肠系膜淋巴液流量,检测其细胞成分及部分生化指标。结果成功建立了大鼠肠系膜淋巴管-颈静脉辅助回流方法,模型维持时间达7d以上。实验组大鼠的肠系膜淋巴液流量多于对照组流量[(2.01±0.12)mL/h vs(0.92±0.09)mL/h,P<0.01]。实验组淋巴液中淋巴细胞数和淋巴细胞百分率高于对照组(P<0.01,P<0.05),而中性粒细胞比例和单核细胞比例均低于对照组(P<0.01)。实验组淋巴液中K^(+)、Na^(+)、CO_(2)和尿素水平均高于对照组(P<0.01),而三酰甘油和P^(3+)水平均低于对照组(P<0.01)。结论肠系膜淋巴管-颈静脉辅助回流技术能够实现大鼠在清醒、不限制饮食和正常活动状态下,实时并长期收集淋巴液,避免了手术应激、全身麻醉或动物受限对实验结果的影响。

Objective To establish a novel sustained collection method for mesenteric lymph fluid by means of assisted reflux from the mesenteric-jugular lymphatic duct in rats.Methods Sixteen male Sprague-Dawley(SD)rats were randomly divided into two groups.The control group underwent duodenal and intestinal lymphatic duct cannulation,after which intestinal lymph fluid was collected.The experimental group underwent jugular vein and intestinal lymphatic duct cannulation to establish intestinal-jugular lymphatic duct assisted reflux.The intestinal lymph fluid was collected on the 7th day after the operation using an awake mobility device.The flow rate of intestinal lymph fluid was recorded,and its cellular components and some biochemical indicators were detected.Results Mesenteric-jugular lymphatic duct vein cannulation assisted reflux was successfully established in rats,and the rat models could be maintained for more than seven days.The intestinal lymph flow rate was(2.01±0.12)mL/h in the experimental group,which was higher than that of the control group[(0.92±0.09)mL/h,P<0.01].The number of lymphocytes(LYM#)and percentage of lymphocytes(LYM/%)in the experimental group were higher than those in the control group(P<0.01).The percentage of neutrophils(NEUT/%)and percentage of monocytes(MONO/%)were lower than those in the control group(P<0.01,P<0.05,respectively).The concentrations of K^(+),Na^(+),CO_(2),and urea in the lymph fluid of the experimental group were higher than those of the control group(P<0.01).However,the concentrations of triacylglycerol(TG)and P^(3+)were lower than those of the control group(P<0.01).Conclusion This novel method can achieve real-time and long-term collection of mesenteric lymph fluid in rats under the condition of being awake,unrestricted in diet and in normal state,avoiding the influence of surgical stress,general anesthesia or animal restraint on the experimental results.