Search PubMed⌕ Search

Biomedical subjects

A A Gruzdkov

Publications and source records attributed to A A Gruzdkov.

At least 19 recordsLinked to original sources
PubMed2006

[The role of facilitated diffusion in glucose transport across the apical membrane of enterocytes].

In chronic experiments on Wistar rats, glucose and galactose absorption in the isolated loop of the small intestine considerably decreased in presence of both phloridzine am phloritine (inhibitors of the glucose transporters SGLT1 and GLUT2). The load of the isolated loop with glucose or galactose solutions scarcely influenced the absorption of 2-deoxi-D-glucose (substrate for GLUT2). According to the immunocytochemical analysis by means of confocal microscopy, after the load of the isolated loop with glucose (75 mM) the labels to GLUT2 and proteinkinase C (PKC betalI) were concentrated mainly in the apical part of the enterocytes, whereas after the load with the Ringer solution--in the basal part of the enterocytes. It was shown on the mathematical model that the part of the facilitated diffusion in the total glucose absorption was considerably lesser in comparison with the active transport mediated by SGLT1. Thus the findings support the hypothesis about a recruitment of the transporter GLUT2 into the apical membrane of the enterocytes and its involvement in glucose transfer across this membrane. However, under natural conditions, the active transport is the main mechanism of glucose absorption, whereas the facilitated diffusion plays a certain role only at high carbohydrate loads.

Animals↗
PubMed2006

[Structural-functional analysis of diffusion in glucose absorption by rat small intestine enterocytes].

To elucidate mechanisms providing transport of sugars across intestinal epithelium, on taking into account the current hypotheses (active transport, participation of paracellular transport and passive component of transcellular transport), it was important to reveal structural changes of tight junctions and distribution of the carriers of facilitated diffusion of GLUT2 and protein kinase C during absorption of glucose. On using confocal and electron microscopy, ultrastructural and immunocytochemical studies of enterocytes after perfusion of isolated rat small intestine fragment with 75 mM glucose (chronic experiment) have shown: 1) fluorescent labels of transporter GLUT2 and PKCbetaII are located in the apical area of enterocytes situated at the upper half of the villus. Antibodies against GLUT2, conjugated with gold, are revealed at the microvilli or apical membrane and in the area of terminal network; 2) no ultrastructural changes of the tight junction are detected on ultrathin sections and freeze--fracture replics. At the same time, fluorescent and gold labels against actin are concentrated in the vicinity of the lateral membrane in the tight junction area. The results obtained can serve a confirmation of a hypothesis that at high glucose concentrations GLUT2 participates in its transfer across the apical membrane.

Animals↗
PubMed2004

[Study of glucose concentrated solution consumption in rats and simulation of glucose distribution along the intestine].

Free ingestion of glucose solution (200 or 400 g/l) by Wistar rats, previously starved for 18-20 Hrs, was investigated in two groups of the animals: with intact small intestine (group 1, n = 9), and a shortened small intestine following the Thiry-Wella isolation of its one third proximal part (group 2, n = 9). In the rats of the group 2, the isolated intestinal loops were perfused in chronic experiments with soulutions of different glucose concentrations to estimate a permeability of the pre-epithelial ("unstirred") layer and "true" kinetic constants of glucose active transport. The rate of glusouse ingestion was found to be 1.3-fold as high in the of rats fgroup 1 than in the rats of group 2 (p < 0.01). According to results of mathematical modeling, the rate of glucose ingestion by rats corresponds to glucose concentration in the initial solutions and to the absorbing capacity of the small intestine due to the substrate regulation of gastric emptying. The model predicts that, during free ingestion by rats of 400 g/l (2200 mM) glucose solution, the substrate concentration in the intestinal lumen under steady state conditions hardly exceeds 75 mM. This fact contradicts a recently proposed hypothesis about a facilitated transport mediated by GLUT2 as the main mechanism of glucose absorption in the small intestine under normal conditions.

Animals↗
PubMed2003

[Kinetic analysis of glycine and glycylglycine absorption in rat small intestine in chronic experiment].

Kinetics of glycylglycine hydrolysis and absorption as well as that of free glycine absorption in isolated loop of the small intestine was studied in chronic experiments in two groups of rats. In the 1st group (n = 5), the isolated loop daily received for 1 or two hours a glucose load (25 mM), whereas in the 2nd group (n = 4)--a glutamic acid load (25 mM). The "true" values (i.e. corrected for the influence of the pre-epithelial layer) of the Michaelis constant for dipeptide transport were lower than those for the free glycine transport: 16 +/- 1.8 versus 36.3 +/- 3.7 mM (in the 1st group) and 15.9 +/- 2.2 versus 34.0 +/- 3.7 mM (in the 2nd group), whereas values of the maximal rate of active transport as calculated per 1 cm of the intestine length were, on the contrary, higher: 0.64 +/- 0.06 versus 0.42 +/- 0.10 mumol/(min.cm) 1st group and 0.86 +/- 0.13 versus 0.56 +/- 0.04 mumol/(min.cm) in the in the 2nd group. It has been shown that, under these conditions, regarded as the most physiological, over 90% of glycylglycine is absorbed via the peptide transport system. Only a small part of this dipeptide amount (less than 10%) splits during membrane hydrolysis with subsequent absorption of the derived glycine. It has also been found that glutamic acid solution as a regular substrate load is more effective (as compared with the glucose solution) in retarding the atrophic changes occurring in the isolated intestine loop and in preserving its structural and functional parameters on a higher level.

Animals↗
PubMed2003

[Structural and functional analysis of glucose adsorption at high maltose concentrations in the rat small intestine in vivo].

To elucidate the mechanism of glucose absorption at high substrate concentrations, we studied structural and ultrastructural peculiarities of enterocytes arranged at different levels along the intestinal villus. The preparations were obtained from an isolated segment of the rat small intestine after its perfusion with maltose solutions with both low (25 mM) and high (100 mM) concentrations, respectively. Under conditions of chronic experiment at high substrate concentration, an enlargement of intercellular clefts, indicating glucose absorption, occurred in deeper areas of the villus. Besides, also in chronic experiment, we studied kinetics of maltose hydrolysis and derived glucose absorption in the isolated segment of the rat small intestine after its perfusion with maltose at superhigh (up to 200 mM) initial concentrations. Based on these data, a conclusion is made that active transport is the main mechanism of absorption of glucose derived from maltose hydrolysis, operating both at low disaccharide concentrations, and in the range of its superhigh (up to 200 mM) concentrations.

Animals↗
PubMed2002

[Kinetic parameters of maltose hydrolysis and glucose intake in the rat small intestine in a chronic experiment].

"True" (corrected for the influence of the pre-epithelial layer) kinetic constants of maltose hydrolysis (Km and Vmax) and Glucose active transport (Kt and Jmax) in the isolated loop of the rat small intestine in chronic experiments were determined using a new mathematical approach. The Km (4.260.25 mM) does not differ from that, obtained in in vitro experiments on the homogenates of mucous membrane taken from the same intestinal loops, and the Vmax (0.72 +/- 0.07 mol/(min.cm)) is 1.7 times lower than that in in vitro experiments. The Kt and Jmax values are 3.18 +/- 0.68 mM and 0.73 +/- 0.07 mol/(min.cm), resp. The estimated values of Km, Kt and Vmax are in accordance with the corresponding published data, whereas the Jmax is several times higher than the value generally believed on the basis of acute experiments in vivo. A high level of glucose absorption in the small intestine of unanesthetized animals is achieved mainly due to a high permeability of the pre-epithelial layer and a high capacity of the active transport as a major mechanism of glucose absorption in the small intestine under normal conditions.

Algorithms↗
PubMed2001

[Mechanisms of glucose absorption at a high carbohydrate level in the rat small intestine in vivo].

At low maltose concentations, the rates of maltose hydrolysis and glucose absorption increased with increasing substrate concentration in the infusate. A significant fluid secretion occurred in the isolated intestinal loop perfused with hypertonic maltose solution, but water fluxes were close to zero in case of almost isotonic perfusion solutions. The findings suggest that the active transport remains a main mechanism of glucose absorption at super-high maltose concentration as well. The rates of maltose hydrolysis and released glucose absorption in the isolated intestinal loops perfused with super-high maltose concentration, enhance mainly due to an increase of effective digestive-absorptive surface of the villi and, in part, to substrate diffusion across the intestinal epithelium rather than paracellular solvent drag.

Animals↗
PubMed1999

Hydrolysis-dependent absorption of disaccharides in the rat small intestine (chronic experiments and mathematical modeling).

In order to throw light on the mechanisms responsible for the enzyme-dependent absorption of disaccharides membrane hydrolysis of maltose and trehalose and the absorption of glucose (free and that derived from disaccharides) were studied in isolated loops (20 cm) of the rat small intestine in chronic experiments. The rates of glucose absorption were 0.26-0.81 micromol x min(-1) x cm(-1) when the loop was perfused with a 12.5 to 75.0 mmol/l free glucose solution, which is only insignificantly higher than the rates observed during perfusion with equivalent maltose solutions. The coupling coefficient (the ratio of glucose absorption rate to the rate of disaccharide hydrolysis) decreased from 0.90 to 0.60 with the increasing maltose concentrations in the infusate from 6.25 to 37.5 mmol/l, but remained unchanged (approximately 0.95) within the same range of trehalose concentrations. The permeability of the pre-epithelial barrier was equivalent to that of unstirred water layer of less than 40 microm thickness. Fluid absorption was within the range of 0.73-2.55 microl x min(-1) x cm(-1), and it showed a correlation with the rates of glucose absorption. The results agree with a model developed on the assumption that free glucose and that released from disaccharides share the same membrane transporters. It could be concluded that a close coupling of disaccharide hydrolysis with derived glucose absorption in chronic experiments is achieved mainly due to a high activity of glucose transporters, which are presumably not associated with membrane disaccharidases. The transcellular active transport is a predominant mechanism of disaccharide-derived glucose absorption under conditions close to physiological.

Absorption↗
PubMed1999

[Effect of protein deficiency in the female rat diet during pregnancy and lactation on the activity of the membrane and soluble forms of digestive enzymes in the offspring small intestine].

Protein deficiency in female rats diet during pregnancy and lactation resulted in deceleration of induction of sucrase both forms in the jejunum and ileum; in acceleration of induction of the maltase membrane from in the jejunum; and in suppression of the lactase membrane form in the ileum; in earlier forming of the adult-type distribution of activity of the membrane form of intestinal alkaline phosphatase and in a decrease in activity of the enzyme soluble form. The findings are corroborated by a suppression of activities of the membrane and soluble forms of the small intestine digestive enzymes in 30-day old rat pups fed with a control (adequate) ration starting 21 days after the birth.

Alkaline Phosphatase↗
PubMed1997

[Analysis of rat enterocyte ultrastructure during glucose absorption].

Electronmicroscopic study of rat enterocytes under glucose load (10-40 mM) has shown some changes of their structure: aggregations of intramembrane particles of the apical membrane in the microvilli region, the dilitation of lateral intercellular spaces below tight junction, the condensation of actin near tight and intermediate junctions. The presence of these changes and almost absolute absence of destructions in tight junctions organization indicate that the main pathway of the isotonic fluid containing glucose across leaky epithelium of rat small intestine is a transcellular one.

Animals↗
PubMed1995

Structural and functional analysis of glucose absorption mechanisms in the rat small intestine in vivo.

The absorption of glucose (free, and released from membrane hydrolysis of maltose) and water in the isolated loop of the rat small intestine was studied in chronic experiments. Even at maximum glucose (75 mmol/l) or maltose (37.5 mmol/l) concentrations the rate of glucose transfer by solvent drag and by diffusion did not exceed 13% and 25%, respectively, of the total rate of glucose absorption. Electron microscopic and immunocytochemical analysis revealed a significant widening of intercellular gaps in the basal epithelium region and an increase of actin density in the vicinity of the tight junctions and between the apical root filaments in enterocytes after glucose or glycine load in chronic and acute experiments. However, very rarely (in 1% of all cases), and only in chronic experiments, structural changes in the tight junctions such as "blisters" and dilatations were also recognised. It is concluded that under normal physiological conditions the absorption of glucose (free, and released from maltose hydrolysis) mainly uses active transport across the apical membrane of the enterocytes.

Animals↗
PubMed1993

[An analysis of the structural characteristics of the tight junction of the enterocytes of the rat small intestine during nutrient absorption (immunoelectron microscopic research)].

To analyze structural changes of intercellular relationships of the enterocytes during glycine, glucose, and triolein absorption the structural and immunocytochemical methods of electron microscopy were used. The study was carried out on the proximal part of the rat small intestine in acute and chronic experiments. In the acute ones glucose or glycine solutions (both of 10 and 40 mM) or triolein emulsion (0.5%) were inserted into the isolated small intestinal segments for 20 min. In chronic experiments the isolated loop of the small intestine was perfused with glucose solution (40 mM). Then the corresponding pieces of the intestinal tissue were fixed for structural and immunocytochemical studies. Rarely (in 1% of all cases), and only in chronic experiments, structural changes in the tight junctions--"blisters" and dilatations--were found. At the same time the analysis of the spatial distribution of actin filaments showed that during glucose and glycine absorption the antiactin tracers were found not only within microvilli and on the "root" filaments but also in vicinity of the tight junction and between terminal filaments. The results obtained agree with the hypothesis about the possibility of paracellular transport of some nutrients induced by sodium-dependent transport of glucose and glycine.

Amino Acids↗
PubMed1993

[The relative role of different mechanisms of glucose absorption in the small intestine under physiological conditions].

Chronic experiments in animals have shown that, even at the maximal glucose (75 mM) and maltose (37.5 mM) concentrations in the initial perfusate, the rate of glucose transfer by a solvent drug does not exceed 10 and 13 per cent, resp. of the total rate of glucose absorption in an isolated intestinal loop. The absorption of glucose under physiological conditions seems to be due mainly to its active transport across the apical membrane of the enterocytes.

Animals↗
PubMed1992

[Trans-sorption as an important mechanism of molecular transport in biological systems].

The mechanism of trans-sorption was shown to be based on mobile adsorption of various substances at specific and unspecific sites localised in spatially organised systems integrated within premembrane structures. Some specifics and possible physiological significance of trans-sorption are discussed using mathematical models of nutrients transfer across autonomous premembrane layer in the small intestine.

Animals↗
PubMed1990

Localization of peptide hydrolysis in the enterocyte and the transport mechanisms of apical membrane.

1. The absence of the effect of anoxia on the hydrolysis rate of a number of dipeptides and one tripeptide by the intact and homogenized mucosa of the small intestine in different mammals (rat, mouse and guinea pig) has been demonstrated. 2. It has been shown that in rats anoxia inhibits intestinal transport both of free glycine and glycine formed during the hydrolysis of Gly-Leu, Leu-Gly, Gly-Pro but not Pro-Gly. 3. Data obtained using the anoxic criterion suggest that the systems of membrane hydrolysis of peptides with the subsequent absorption of released amino acids presents a dominant mechanism of protein assimilation under normal physiological conditions. 4. However, they do not exclude the possibility of the transport of peptides across the apical membrane of enterocytes.

Absorption↗