Isolation and Characterization of a New Major Intestinal CYP3A Form, CYP3A62, in the Rat

Matsubara, Tsutomu; Kim, Hye-Ji; Miyata, Masaaki; Shimada, M.; Nagata, Kiyoshi; Yamazoe, Yasushi

doi:10.1124/jpet.103.061671

Cited by 88 publications

(68 citation statements)

References 35 publications

(41 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This finding is consistent with data published by Matsubara et al (2004), Takara et al (2003), and Takemoto et al (2003) showing decreasing expression of CYP3A mRNA and decreasing activity along the whole intestine. Even for the human equivalent CYP3A4 declining mRNA expression and activity profile along the human intestine could be demonstrated (Thummel et al, 1997).…”

Section: Mitschke Et Alsupporting

confidence: 83%

Characterization of Cytochrome P450 Protein Expression along the Entire Length of the Intestine of Male and Female Rats

Mitschke

Reichel²,

Fricker³

et al. 2008

Drug Metab Dispos

View full text Add to dashboard Cite

ABSTRACT:Intestinal cytochrome P450 (P450) proteins play an important role in the biotransformation of drugs and may significantly limit their oral absorption and bioavailability. Therefore, we have investigated the amount of P450 proteins via Western blot analysis along the entire intestine of male and female rats. Despite of the use of an inbred rat strain, controlled housing conditions for the animals, and a timed sample preparation, high interindividual differences in the expression of all P450 proteins was observed. CYP3A (135-243 fmol/mg of protein) and CYP2B1 (107-645 fmol/mg of protein) were the most abundant P450 isoforms in the duodenum and jejunum of rat intestine but were present in neither the ileum nor the colon. Compared with CYP2B1 and CYP3A, CYP2D1 (25-71 fmol/mg of protein) and CYP2C6 (3-10 fmol/mg of protein) were only expressed in minor amounts. CYP2C11 could not be identified in the entire rat intestine. In conclusion, this is the first systematic evaluation and quantification of the expression of P450 proteins along the entire length of the intestine in both male and female rats. These data will provide a basis for a better understanding of the extent of intestinal metabolism along the gastrointestinal tract.Absorption through the gut is a key step in the oral delivery of drugs. The main interface between gut lumen and the bloodstream is an epithelial cell layer consisting of polarized enterocytes controlling the passage of exogenous substances into the portal circulation. Enterocytes have a variety of structural features, including tight junctions reducing paracellular permeability, numerous drug transporters, and a set of metabolic enzymes that may all affect the entry of drugs into the body. The extent to which a drug is absorbed also depends on the intrinsic properties of the compound such as solubility, permeability, efflux or uptake transport properties, and susceptibility to metabolic degradation (Martinez and Amidon, 2002;Benet et al., 2004).Although the liver is known as the major site of first-pass extraction, recent studies have indicated that the small intestine also contributes significantly to the first-pass metabolism of many drugs, e.g., cyclosporine (Wu et al., 1995), nifedipine (Iwao et al., 2002), midazolam (Paine et al., 1996, and diltiazem (Iwao et al., 2004). It is known that several uptake and efflux transporter and P450 isoforms are expressed in the human and rat intestine (van de Kerkhof et al., 2007). If a drug is a substrate of efflux transporters, it may enter and exit the enterocytes several times, and, with each cycle, small quantities may be metabolized by cytochrome P450 (P450) enzymes localized in the endoplasmic reticulum. The P450 enzymes belong to a superfamily of heme proteins that show a broad substrate specificity, with substrates ranging in size from ethylene (M r 28) to cyclosporine (M r 1201) (Isin and Guengerich, 2007). Although the liver is regarded as the main organ of drug metabolism, P450 proteins are also expressed in other tissues, e.g., kidn...

show abstract

Section: Mitschke Et Alsupporting

confidence: 83%

Characterization of Cytochrome P450 Protein Expression along the Entire Length of the Intestine of Male and Female Rats

Mitschke

Reichel²,

Fricker³

et al. 2008

Drug Metab Dispos

View full text Add to dashboard Cite

show abstract

“…administration of CyA, the disposition kinetics of CyA was little changed by LTX treatment (LTX 8 mg W kg W day, p.o., for 3 weeks), presumably because the expression of mdr1 and CYP3A mRNAs in the liver did not change almost. Moreover, Matsubara et al 21) reported that the mRNAs and proteins of CYP3A1 and CYP3A2 were induced in the rat liver after the DEX treatment (100 mg W kg W day) for 3 days, while those of CYP3A62 and CYP3A9 were induced in the intestine. In this study, although we did not evaluate the eŠect of LTX treatment on the expression of CYP3A62 and CYP3A9, we believe that the in‰uence of LTX treatment on these proteins was slight, if any, because the CLtot value of CyA after the i.v.…”

Section: Discussionmentioning

confidence: 99%

Long-term Levothyroxine Treatment Decreases the Oral Bioavailability of Cyclosporin A by Inducing P-glycoprotein in Small Intestine

Jin

Shimada

Shintani

et al. 2005

Drug Metabolism and Pharmacokinetics

View full text Add to dashboard Cite

Summary:We have noticed that the trough level of blood concentration of cyclosporin A (CyA) tends to be lower in patients receiving long-term oral levothyroxine (LTX) than in patients not receiving LTX. We conˆrmed this clinical observation in experiments using Wistar rats orally given LTX (8 mg W kg) or saline (control) for 3 weeks, followed by CyA (10 mg W kg). The LTX treatment had little eŠect on the blood concentrations of CyA after i.v. administration, whereas they were decreased signiˆcantly after p.o. administration. After p.o. administration, the value of the area under the blood concentration-time curve from 0 to 24 hr and the bioavailability of CyA in the LTX group were decreased to only about onefth and a quarter of those in the control group, respectively. After treatment with LTX, the expression levels of mdr1a, mdr1b and CYP3A2 mRNAs in the duodenum were markedly increased to about twice the control, but in jejunum, ileum and liver the expression levels were little changed. Theseˆndings suggest that the absorption of CyA, which occurs mainly from the upper intestine, is reduced as a result of eOEux transport via P-glycoprotein induced by LTX. In conclusion, careful monitoring of CyA levels is required in the event of LTX administration to patients receiving immunotherapy with CyA.

show abstract

“…Mouse hepatic and small intestinal microsomes were prepared according to the method reported previously (Matsubara et al, 2004), separated by SDS-polyacrylamide gel electrophoresis, and transferred to a nitrocellulose membrane. The membrane was immunostained with the anti-CYP3A antibody (Kawano et al, 1987) and alkaline phosphatase-conjugated goat anti-rabbit IgG, and signals were visualized with 5-bromo-4-chloro-3-indolylphosphate and nitro blue tetrazolium.…”

Section: Methodsmentioning

confidence: 99%

Role of Vitamin D Receptor in the Lithocholic Acid-Mediated CYP3A Induction in Vitro and in Vivo

Matsubara¹,

Yoshinari²,

Aoyama³

et al. 2008

Drug Metab Dispos

View full text Add to dashboard Cite

ABSTRACT:Lipophilic bile acids are suggested to be involved in the endogenous expression of CYP3A4 in human and experimental animals as ligands of nuclear receptors. To verify the nuclear receptor specificity, the bile acid-mediated induction of CYP3A4 has been studied in vitro and in vivo in the present study. Lithocholic acid (LCA) strongly enhanced the activities of the CYP3A4 reporter gene, which contained multiple nuclear receptor binding elements, in both HepG2 and LS174T cells. The introduction of small interfering RNA for human vitamin D receptor (VDR), but not for human pregnane X receptor, reduced the LCA-induced activation of the reporter gene in these cells, suggesting the major role of VDR in the LCA induction of CYP3A4. Consistently, oral administration of LCA (100 mg/kg/day for 3 days) increased Cyp3a protein levels in the intestine but not in the liver, where a negligible level of VDR mRNA is detected. The selective role of VDR was tested in mice with the adenoviral overexpression of the receptor. Oral administration of LCA had no clear influence on the CYP3A4 reporter activity in the liver of control mice. In mice with the adenovirally expressed VDR, LCA treatment (100 or 400 mg/kg/day for 3 days) resulted in the enhanced reporter activities and increased levels of Cyp3a proteins in the liver. These results indicate the selective involvement of VDR, but not pregnane X receptor, in the LCA-mediated induction of both human and mouse CYP3As in vivo.

show abstract

Isolation and Characterization of a New Major Intestinal CYP3A Form, CYP3A62, in the Rat

Cited by 88 publications

References 35 publications

Characterization of Cytochrome P450 Protein Expression along the Entire Length of the Intestine of Male and Female Rats

Characterization of Cytochrome P450 Protein Expression along the Entire Length of the Intestine of Male and Female Rats

Long-term Levothyroxine Treatment Decreases the Oral Bioavailability of Cyclosporin A by Inducing P-glycoprotein in Small Intestine

Role of Vitamin D Receptor in the Lithocholic Acid-Mediated CYP3A Induction in Vitro and in Vivo

Contact Info

Product

Resources

About