Influence of infusate viscosity on intestinal absorption in the rat. An explanation of previous discrepant results.

Previous studies of the influence of increased luminal viscosity on intestinal absorption have yielded conflicting results ranging from no effect to a marked diminution. We measured the absorption of three probes (carbon monoxide, [14C]warfarin, 5.5 mM glucose) from a saline infusate or from saline containing 0.6% guar, which yielded a 20-fold increase in viscosity. Two animal models were used: (a) conscious nonlaparotomized rats with chronically implanted cannulas and (b) anesthetized laparotomized rats. In the anesthetized laparotomized rats, absorption was independent of perfusate viscosity. In the conscious nonlaparotomized rats, the absorption of each of the three probes was significantly greater than in the anesthetized laparotomized rats and increased viscosity caused a 60%-70% decrease in the clearance of the three probes. In anesthetized laparotomized rats, we have shown that fluid moves with laminar flow, and increased infusate viscosity cannot further reduce luminal stirring (or absorption). In conscious, nonlaparotomized rats, laminar flow is disrupted by normal gut motility causing better luminal stirring. Such stirring is inhibited by a viscous infusate resulting in decreased absorption. We conclude that the conflicting results seen in previous studies can be attributed to the model used. In conscious animals where luminal stirring was good, a viscous infusate caused decreased absorption.

Influence of shaking on peritoneal transfer in rats.

To determine the role played by stagnant peritoneal fluid layers in the diffusion of solutes between peritoneal cavity and blood, we measured peritoneal transfer of urea, creatinine, [14C]-L-glucose and protein in anesthetized rats shaken at varying rates on an orbital platform shaker. The diffusion transfer rates of the low molecular weight solutes increased dramatically with shaking, with near maximal values obtained at a shaking rate of 250 RPM. The permeability area product (PA) for each of the low molecular weight solutes increased about fourfold with rapid shaking while the PA of protein increased by only about 50%. It seems likely while the PA of protein increased by only about 50%. It seems likely that shaking increased PA primarily via reduction of the thickness of stagnant peritoneal fluid layers, although increases in surface area or changes in tissue permeability cannot be excluded with certainty. We conclude that stagnant fluid layers probably are the rate limiting step in diffusive peritoneal transfer of low molecular weight solutes in stationary rats.

Physiological measurement of luminal stirring in perfused rat jejunum.

Poor stirring of intestinal contents yields a preepithelial diffusion barrier that is thought to be the rate-limiting step in absorption of many compounds. In many previous studies, the resistance of this barrier is equal to an unstirred water layer of 300-800 micron. Using three probes, CO, glucose, and [14C]warfarin, we measured the preepithelial resistance in a 30-cm segment in rats that were 1) conscious, 2) anesthetized with pentobarbital sodium, or 3) anesthetized and laparotomized. Measurements with each of the probes showed that the maximal preepithelial resistance in conscious rats was equivalent to an unstirred layer of only approximately 100 micron. Anesthesia roughly doubled this resistance, and anesthesia and laparotomy caused a sixfold increase (unstirred layer of approximately 600 micron). We conclude that the luminal stirring is much more efficient than previously has been appreciated. The very thick jejunal unstirred layers reported previously (300-800 micron) reflect the results of studies performed under nonphysiological conditions or studies employing inappropriate techniques to measure luminal stirring.

Use of laminar flow and unstirred layer models to predict intestinal absorption in the rat.

Carbon monoxide (CO) and [14C]warfarin were used to measure the preepithelial diffusion resistance resulting from poor luminal stirring (RL) in the constantly perfused rat jejunum at varying degrees of distension (0.05, 0.1, and 0.2 ml/cm). RL was much greater than epithelial cell resistance, indicating that poor stirring was the limiting factor in absorption and that an appropriate model of stirring should accurately predict absorption. A laminar flow model accurately predicted the absorption rate of both probes at all levels of gut distension, as well as the absorption of glucose when RL was the rate-limiting factor in absorption. In contrast, an unstirred layer model would not have predicted that gut distension would have little influence on absorption, and would have underestimated [14C]warfarin absorption relative to CO. We concluded that in the perfused rat jejunum, laminar flow accurately models luminal stirring and an unstirred layer should be considered to be a unit of resistance in laminar flow, rather than a model of luminal stirring.

Storage of breath samples for H2 analyses.

Breath hydrogen tests are limited by leak of gases from breath samples from standard glass or plastic syringes. These losses may be substantial when these samples are stored for long periods of time. We describe a simple manipulation involving the use of a mineral oil or water seal at the end of the syringe barrel that markedly reduces the leakage of H2 and other gases to negligible levels.

Quantitative assessment of luminal stirring in the perfused small intestine of the rat.

We measured the resistance (RL) to CO absorption that resulted from poor luminal stirring in the constantly perfused rat jejunum. RL or calculated unstirred layer thickness was greater for 30-cm than 10-cm long segments, indicating lack of a uniform thickness of unstirred layer. The possibility that laminar flow existed in the gut was first tested by calculating expected CO absorptions from fluid moving with laminar flow. These values agreed closely with observed absorption rates. Laminar flow also was supported by the observation that CO absorption was independent of perfusate viscosity. Lastly, after sudden addition of phenolsulfonphthalein (PSP) to the perfusate, PSP outflow concentration was similar in tygon tubing (which has laminar flow) and a gut segment of comparable dimension. We conclude that flow in the perfused gut is laminar and that this laminar flow has many implications for studies carried out with the constant-perfusion technique.