Trevaskis Natalie L, Hu Luojuan, Caliph Suzanne M, Han Sifei, Porter Christopher J H
Drug Delivery, Disposition, and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus);
Drug Delivery, Disposition, and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus).
J Vis Exp. 2015 Mar 6(97):52389. doi: 10.3791/52389.
The intestinal lymphatic system plays key roles in fluid transport, lipid absorption and immune function. Lymph flows directly from the small intestine via a series of lymphatic vessels and nodes that converge at the superior mesenteric lymph duct. Cannulation of the mesenteric lymph duct thus enables the collection of mesenteric lymph flowing from the intestine. Mesenteric lymph consists of a cellular fraction of immune cells (99% lymphocytes), aqueous fraction (fluid, peptides and proteins such as cytokines and gut hormones) and lipoprotein fraction (lipids, lipophilic molecules and apo-proteins). The mesenteric lymph duct cannulation model can therefore be used to measure the concentration and rate of transport of a range of factors from the intestine via the lymphatic system. Changes to these factors in response to different challenges (e.g., diets, antigens, drugs) and in disease (e.g., inflammatory bowel disease, HIV, diabetes) can also be determined. An area of expanding interest is the role of lymphatic transport in the absorption of orally administered lipophilic drugs and prodrugs that associate with intestinal lipid absorption pathways. Here we describe, in detail, a mesenteric lymph duct cannulated rat model which enables evaluation of the rate and extent of lipid and drug transport via the lymphatic system for several hours following intestinal delivery. The method is easily adaptable to the measurement of other parameters in lymph. We provide detailed descriptions of the difficulties that may be encountered when establishing this complex surgical method, as well as representative data from failed and successful experiments to provide instruction on how to confirm experimental success and interpret the data obtained.
肠道淋巴系统在液体运输、脂质吸收和免疫功能中发挥着关键作用。淋巴液通过一系列淋巴管和淋巴结直接从小肠流出，这些淋巴管和淋巴结在肠系膜上淋巴管处汇合。因此，对肠系膜淋巴管进行插管能够收集从小肠流出的肠系膜淋巴液。肠系膜淋巴液由免疫细胞的细胞部分（99%为淋巴细胞）、水性部分（液体、肽和蛋白质，如细胞因子和肠道激素）和脂蛋白部分（脂质、亲脂性分子和载脂蛋白）组成。因此，肠系膜淋巴管插管模型可用于测量一系列物质从肠道经淋巴系统运输的浓度和速率。还可以确定这些物质在应对不同挑战（如饮食、抗原、药物）以及疾病（如炎症性肠病、HIV、糖尿病）时的变化。一个日益受到关注的领域是淋巴运输在口服亲脂性药物和前体药物吸收中的作用，这些药物与肠道脂质吸收途径相关。在此，我们详细描述了一种肠系膜淋巴管插管大鼠模型，该模型能够在肠道给药后的数小时内评估脂质和药物经淋巴系统运输的速率和程度。该方法易于适用于测量淋巴中的其他参数。我们详细描述了建立这种复杂手术方法时可能遇到的困难，以及失败和成功实验的代表性数据，以指导如何确认实验成功并解释所获得的数据。
