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J Surg Res. 2015 Aug;197(2):225-30. doi: 10.1016/j.jss.2015.02.049. Epub 2015 Apr 20.

A new method to measure intestinal secretion using fluorescein isothiocyanate-inulin in small bowel of rats.

Author information

1
Department of Surgery, Yale University School of Medicine, New Haven, Connecticut.
2
Department of Surgery, Yale University School of Medicine, New Haven, Connecticut. Electronic address: john.geibel@yale.edu.

Abstract

BACKGROUND:

Small intestine ischemia can be seen in various conditions such as intestinal transplantation. To further understand the pathologic disruption in ischemia-reperfusion injury, we have developed a method to measure fluid changes in the intestinal lumen of rats.

METHODS:

Two 10-cm rat intestine segments were procured, connected to the terminal apertures of a perfusion device, and continuously infused with 3 mL of HEPES solution (control solution) containing 50 μM of fluorescein isothiocyanate (FITC)-inulin. The perfusion device consists of concentric chambers that contain the perfused bowel segments, which are maintained at 37°C via H₂O bath. The individual chamber has four apertures as follows: two fill and/or drain the surrounding HEPES solution on the blood side of the tissue. The others provide flow of HEPES solution containing FITC-inulin through the lumens. The experimental intestine was infused with the same solution with 100 μM of Forskolin. A pump continuously circulated solutions at 6 mL/min. Samples were collected at 15-min intervals until 150 min and were measured by the nanoflourospectrometer.

RESULTS:

A mean of 6-μM decrease in the FITC-inulin concentration in the Forskolin-treated experimental intestine was observed in comparison with that in the control intestine. The FITC-inulin count dilution in the experimental intestine is a result of an increase of fluid secretion produced by the effect of Forskolin, with P values <0.0001.

CONCLUSIONS:

We demonstrate that it is possible to measure luminal fluid changes over time using our new modified perfusion system along with FITC-inulin to allow real-time determinations of fluid and/or electrolyte movement along the small intestine.

KEYWORDS:

Cyclic nucleotides; Fluid secretion; Fluorescence; Forskolin; Intestinal transplant; Ischemia

PMID:
25976849
DOI:
10.1016/j.jss.2015.02.049
[Indexed for MEDLINE]

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