In Situ Perfusion Model in Rat Colon for Drug Absorption Studies: Comparison with Small Intestine and Caco-2 Cell Model

2015 ◽  
Vol 104 (9) ◽  
pp. 3136-3145 ◽  
Author(s):  
Isabel Lozoya-Agullo ◽  
Isabel González-Álvarez ◽  
Marta González-Álvarez ◽  
Matilde Merino-Sanjuán ◽  
Marival Bermejo
Keyword(s):  
Small Intestine ◽  
Drug Absorption ◽  
Cell Model ◽  
Rat Colon ◽  
In Situ Perfusion ◽  
Perfusion Model ◽  
Absorption Studies

scholarly journals Generation of tetracycline-controllable CYP3A4-expressing Caco-2 cells by the piggyBac transposon system

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Moe Ichikawa ◽  
Hiroki Akamine ◽  
Michika Murata ◽  
Sumito Ito ◽  
Kazuo Takayama ◽  
...  
Keyword(s):  
Small Intestine ◽  
Drug Absorption ◽  
Cell Model ◽  
Negative Effects ◽  
Piggybac Transposon ◽  
Drug Metabolizing Enzyme ◽  
Metabolizing Enzyme ◽  
Cyp3a4 Expression

AbstractCaco-2 cells are widely used as an in vitro intestinal epithelial cell model because they can form a monolayer and predict drug absorption with high accuracy. However, Caco-2 cells hardly express cytochrome P450 (CYP), a drug-metabolizing enzyme. It is known that CYP3A4 is the dominant drug-metabolizing enzyme in human small intestine. In this study, we generated CYP3A4-expressing Caco-2 (CYP3A4-Caco-2) cells and attempted to establish a model that can simultaneously evaluate drug absorption and metabolism. CYP3A4-Caco-2 cells were generated by piggyBac transposon vectors. A tetracycline-controllable CYP3A4 expression cassette (tet-on system) was stably transduced into Caco-2 cells, thus regulating the levels of CYP3A4 expression depending on the doxycycline concentration. The CYP3A4 expression levels in CYP3A4-Caco-2 cells cultured in the presence of doxycycline were similar to or higher than those of adult small intestine. The CYP3A4-Caco-2 cells had enough ability to metabolize midazolam, a substrate of CYP3A4. CYP3A4 overexpression had no negative effects on cell proliferation, barrier function, and P-glycoprotein activity in Caco-2 cells. Thus, we succeeded in establishing Caco-2 cells with CYP3A4 metabolizing activity comparable to in vivo human intestinal tissue. This cell line would be useful in pharmaceutical studies as a model that can simultaneously evaluate drug absorption and metabolism.

The Suitability of an in Situ Perfusion Model for Permeability Determinations:  Utility for BCS Class I Biowaiver Requests

2006 ◽  
Vol 3 (6) ◽  
pp. 686-694 ◽  
Author(s):  
Jae-Seung Kim ◽  
Stefanie Mitchell ◽  
Paul Kijek ◽  
Yasuhiro Tsume ◽  
John Hilfinger ◽  
...  
Keyword(s):  
Class I ◽  
In Situ Perfusion ◽  
Perfusion Model

Comparison of segmental-dependent permeability in human and in situ perfusion model in rat

2017 ◽  
Vol 107 ◽  
pp. 191-196 ◽  
Author(s):  
Alejandro Ruiz-Picazo ◽  
Isabel Lozoya-Agullo ◽  
Miguel Ortiz-Azcarate ◽  
Matilde Merino-Sanjuán ◽  
Marta González-Álvarez ◽  
...  
Keyword(s):  
In Situ Perfusion ◽  
Perfusion Model

Determination of intestinal permeability using in situ perfusion model in rats: Challenges and advantages to BCS classification applied to digoxin

2018 ◽  
Vol 551 (1-2) ◽  
pp. 148-157 ◽  
Author(s):  
Tamires Guedes Caldeira ◽  
Alejandro Ruiz-Picazo ◽  
Isabel Lozoya-Agullo ◽  
Dênia Antunes Saúde-Guimarães ◽  
Marta González-Álvarez ◽  
...  
Keyword(s):  
Intestinal Permeability ◽  
In Situ Perfusion ◽  
Perfusion Model

scholarly journals Selective evaluation of high density lipoprotein from mouse small intestine by an in situ perfusion technique

2014 ◽  
Vol 55 (5) ◽  
pp. 905-918 ◽  
Author(s):  
Satoshi Yamaguchi ◽  
Bo Zhang ◽  
Takeshi Tomonaga ◽  
Utako Seino ◽  
Akiko Kanagawa ◽  
...  
Keyword(s):  
Small Intestine ◽  
High Density Lipoprotein ◽  
Density Lipoprotein ◽  
High Density ◽  
Perfusion Technique ◽  
In Situ Perfusion ◽  
Mouse Small Intestine

Neurokinin A increases duodenal mucosal permeability, bicarbonate secretion, and fluid output in the rat

1997 ◽  
Vol 273 (5) ◽  
pp. G1077-G1086 ◽  
Author(s):  
Anneli Hällgren ◽  
Gunnar Flemström ◽  
Per M. Hellström ◽  
Mikael Lördal ◽  
Sandra Hellgren ◽  
...  
Keyword(s):  
Nicotinic Receptors ◽  
Receptor Activation ◽  
Neurokinin A ◽  
Bicarbonate Secretion ◽  
Inhibitory Effects ◽  
Mucosal Permeability ◽  
Duodenal Motility ◽  
In Situ Perfusion ◽  
Perfusion Model

The aim of this study was to examine the integrative response to neurokinin A (NKA) on duodenal mucosal permeability, bicarbonate secretion, fluid flux, and motility in an in situ perfusion model in anesthetized rats. Intravenous infusion of NKA (100, 200, and 400 pmol ⋅ kg−1 ⋅ min−1) induced duodenal motility. Furthermore, duodenal mucosal bicarbonate secretion, fluid output, and mucosal permeability increased in response to NKA. Pretreatment with the nicotinic antagonist hexamethonium did not change the response in any of the parameters investigated, whereas the NK2-receptor antagonist MEN 10,627 effectively inhibited all responses to NKA. Indomethacin induced duodenal motility and stimulated bicarbonate secretion. In indomethacin-treated rats, NKA further increased motility but decreased indomethacin-stimulated bicarbonate secretion by 70%. The NKA-induced increase in mucosal permeability was unaltered by indomethacin. It is concluded that NKA not only induces motility but also increases mucosal permeability and fluid output. Furthermore, the neuropeptide may have both stimulative and inhibitory effects on bicarbonate secretion. All responses to NKA are dependent on NK-2 receptor activation but are not mediated through nicotinic receptors.

scholarly journals The Advantages of the Ussing Chamber in Drug Absorption Studies

2005 ◽  
Vol 10 (5) ◽  
pp. 517-523 ◽  
Author(s):  
Yasumasa Gotoh ◽  
Noboru Kamada ◽  
Denichi Momose
Keyword(s):  
Small Intestine ◽  
Membrane Permeability ◽  
Drug Absorption ◽  
Ussing Chamber ◽  
In Vivo Studies ◽  
Glycoprotein P ◽  
Absorption Studies ◽  
High Concentrations ◽  
Absorption Percentage

By adding high concentrations of test drugs to an Ussing chamber with rat jejunum, we established a systemthat yields very high correlations between the rat absorption percentage and the membrane permeability, and that can accurately predict the absorption percentage for rats. An advantage of this technique is that, unlike the results obtained using Caco-2, the slope of the absorption/membrane-permeability curve is gentle, which facilitates a more exact prediction of the absorption percentage. In addition, the results obtained with this technique demonstrated that it could be used to evaluate the absorption percentage of drugs with an affinity for P-glycoprotein (P-gp), which cannot be assessed using Caco-2. Thismethod also allows for cassette screening, whichwould facilitate evaluation of the contribution of P-gp to absorption in the small intestine. Cassette screening showed that absorption of fexofenadinewas unaffected by combinationwith the P-gp substrate ketoconazole. Consistent with this finding, in vivo studies showed that ketoconazole did not affect the Fa Fg for fexofenadine, a pharmacokinetic parameter that reflects absorption and bioavailability in the small intestine. This confirms the usefulness of the Ussing chamber for cassette screening and also suggests that intestinal P-gp has a minimal contribution to drug absorption.

scholarly journals Part of quercetin absorbed in the small intestine is conjugated and further secreted in the intestinal lumen

1999 ◽  
Vol 277 (1) ◽  
pp. G120-G126 ◽  
Author(s):  
Vanessa Crespy ◽  
Christine Morand ◽  
Claudine Manach ◽  
Catherine Besson ◽  
Christian Demigne ◽  
...  
Keyword(s):  
Small Intestine ◽  
Intestinal Wall ◽  
Intestinal Lumen ◽  
Intestinal Cells ◽  
Biliary Duct ◽  
In Situ Perfusion ◽  
Perfusion Period ◽  
Hydrolysis Of

Rutin and quercetin absorption and metabolism were investigated in rats after in situ perfusion of jejunum plus ileum (15 nmol/min). In contrast to rutin, a high proportion of quercetin (two-thirds) disappeared during perfusion, reflecting extensive transfer into the intestinal wall. Net quercetin absorption was not complete (2.1 nmol/min), inasmuch as 52% were reexcreted in the lumen as conjugated derivatives (7.7 nmol/min). Enterohepatic recycling contribution of flavonoids was excluded by catheterization of the biliary duct before perfusion. After a 30-min perfusion period, 0.71 μM of quercetin equivalents were detected in plasma, reflecting a significant absorption from the small intestine. The differential hydrolysis of effluent samples by glucuronidase and/or sulfatase indicates that the conjugated forms released in the lumen were 1) glucuronidated derivatives of quercetin and of its methoxylated forms (64%) and 2) sulfated form of quercetin (36%). In vitro quercetin glucuronides synthetized using jejunal and ileal microsomal fractions were similar to those recovered in the effluent of perfusion. These data suggest that glucuronidation and sulfatation take place in intestinal cells, whereas no glucurono-sulfoconjugates could be detected in the effluent. The present work shows that a rapid quercetin absorption in the small intestine is very effective together with its active conjugation in intestinal cells.

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