An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products
An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products
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DOI：
10.3791/58456
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发表时间：
2018-10-01
影响因子：
1.2
通讯作者：
Ufnal, Marcin
中科院分区：
文献类型：
--
作者：
Jaworska, Kinga;Huc, Tomasz;Ufnal, Marcin
肠血屏障（GBB）控制着营养物质、细菌代谢产物和药物从肠腔进入血液的通道。GBB的完整性在胃肠道、心血管和代谢疾病中受到干扰，这可能导致生物活性化合物（如肠道细菌代谢物）更容易进入血流。因此，GBB的渗透性可能是肠道和肠外疾病的标志物。此外，细菌代谢产物的渗透增加可能会影响整个生物体的功能。这些方法的准确性是有限的，因为GBB的功能取决于肠道血流。另一方面，常用的体内方法可能因肝脏和肾脏性能而存在偏倚，因为这些方法是基于对外源性标志物的尿液或/和外周血浓度的评价。在这里，我们提出了一个直接测量GBB渗透性在大鼠使用的基础上，在体内的方法门静脉采血，保留肠血流量，几乎不受肝脏和肾脏功能。聚氨酯导管插入门静脉和下腔静脉肝静脉汇合。在基线时和在将选定的标记物施用到胃肠道的期望部分中之后对血液取样。在这里，我们提出了该方法的几个应用，包括（1）评价结肠对TMA（一种肠道细菌代谢物）的渗透性，（2）评价TMA的肝脏清除率，以及（3）评价肠道细菌衍生的短链脂肪酸的肠道-门静脉血-肝脏-外周血途径。此外，该方案还可用于跟踪药物的肠吸收和肝脏代谢或用于测量门静脉血压。
The gut-blood barrier (GBB) controls the passage of nutrients, bacterial metabolites and drugs from intestinal lumen to the bloodstream. The GBB integrity is disturbed in gastrointestinal, cardiovascular and metabolic diseases, which may result in easier access of biologically active compounds, such as gut bacterial metabolites, to the bloodstream. Thus, the permeability of the GBB may be a marker of both intestinal and extraintestinal diseases. Furthermore, the increased penetration of bacterial metabolites may affect the functioning of the entire organism.Commonly used methods for studying the GBB permeability are performed ex vivo. The accuracy of those methods is limited, because the functioning of the GBB depends on intestinal blood flow. On the other hand, commonly used in vivo methods may be biased by liver and kidney performance, as those methods are based on evaluation of urine or/and peripheral blood concentrations of exogenous markers. Here, we present a direct measurement of GBB permeability in rats using an in vivo method based on portal blood sampling, which preserves intestinal blood flow and is virtually not affected by the liver and kidney function.Polyurethane catheters are inserted into the portal vein and inferior vena cava just above the hepatic veins confluence. Blood is sampled at baseline and after administration of a selected marker into a desired part of the gastrointestinal tract. Here, we present several applications of the method including (1) evaluation of the colon permeability to TMA, a gut bacterial metabolite, (2) evaluation of liver clearance of TMA, and (3) evaluation of a gut-portal blood-liver-peripheral blood pathway of gut bacteria-derived short-chain fatty acids. Furthermore, the protocol may also be used for tracking intestinal absorption and liver metabolism of drugs or for measurements of portal blood pressure.