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Intestinal and hepatic expression of cytochrome P450s and mdr1a in rats with indomethacin-induced small intestinal ulcers.

Kawauchi S, Nakamura T, Yasui H, Nishikawa C, Miki I, Inoue J, Horibe S, Hamaguchi T, Tanahashi T, Mizuno S - Int J Med Sci (2014)

Bottom Line: Gene expression of the CYP family of enzymes and mdr1a was measured with quantitative real-time polymerase chain reaction (qPCR).Although previous studies have shown a direct effect of INM on CYP3A activity, we could not confirm any change in hepatic CY3A4 expression (major isoform of human CYP3A) in vitro.INM-induced SIUs had a subtle effect on intestinal CYP expression, but had an apparent action on hepatic CYP, which was influenced, at least in part, by the secondary inflammation.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Medical Pharmaceutics, Kobe Pharmaceutical University, 4-19-1 Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan; ; 2. Educational Center for Clinical Pharmacy, Kobe Pharmaceutical University, 4-19-1 Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan;

ABSTRACT

Background: Non-steroidal anti-inflammatory drugs induce the serious side effect of small intestinal ulcerations (SIUs), but little information is available regarding the consequences to drug metabolism and absorption.

Aim: We examined the existence of secondary hepatic inflammation in rats with indomethacin (INM)-induced SIUs and assessed its relationship to the cytochrome P450 (CYP) and P-glycoprotein (mdr1a), the major drug-metabolizing factors in the small intestine and the liver.

Methods: Gene expression of the CYP family of enzymes and mdr1a was measured with quantitative real-time polymerase chain reaction (qPCR). Vancomycin (VCM), a poorly absorbed drug, was administered intraduodenally to rats with SIUs.

Results: INM induced SIUs predominantly in the lower region of the small intestine with high expression of inflammatory markers. Liver dysfunction was also observed, which suggested a secondary inflammatory response in rats with SIUs. In the liver of rats with SIUs, the expression of CYP2C11, CYP2E1, and CYP3A1 was significantly decreased, and loss of CYP3A protein was observed. Although previous studies have shown a direct effect of INM on CYP3A activity, we could not confirm any change in hepatic CY3A4 expression (major isoform of human CYP3A) in vitro. The plasma VCM concentration was increased in rats with SIUs due to partial absorption from the mucosal injury, but not in normal mucosa.

Conclusions: INM-induced SIUs had a subtle effect on intestinal CYP expression, but had an apparent action on hepatic CYP, which was influenced, at least in part, by the secondary inflammation. Furthermore, drug absorption was increased in rats with SIUs.

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Changes in the mRNA expression of CYPs and mdr1a in the rat small intestine with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. Total RNA was extracted from the upper, middle and lower small intestine. Expression levels of CYP and mdr1a mRNA were determined by real-time PCR, as described in the Materials and Methods. The data for mRNA expression for the non-SIUs (open triangles) and SIU groups (closed triangles) are expressed as the ratio of the mean value for each target gene in the upper region of the small intestine of the control group (open circles). Points represent the individual data, and bars represent the mean value for 5 to 6 rats per group. *P < 0.05, statistically significant.
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Figure 4: Changes in the mRNA expression of CYPs and mdr1a in the rat small intestine with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. Total RNA was extracted from the upper, middle and lower small intestine. Expression levels of CYP and mdr1a mRNA were determined by real-time PCR, as described in the Materials and Methods. The data for mRNA expression for the non-SIUs (open triangles) and SIU groups (closed triangles) are expressed as the ratio of the mean value for each target gene in the upper region of the small intestine of the control group (open circles). Points represent the individual data, and bars represent the mean value for 5 to 6 rats per group. *P < 0.05, statistically significant.

Mentions: Figure 4 shows the effect of INM on the relative mRNA expression of CYPs and mdr1a in the upper, middle, and lower regions of the small intestine. In control rats, all CYP expression levels measured were significantly higher in the upper region than in the lower region (P < 0.05). An inverse trend was observed to expression of mdr1a, but it was not significant (P = 0.0503). In rats with non-SIU and SIU tissues, CYP and mdr1a expression were similar to those observed in control rats, with relatively high expression in the upper region. In particular, there was significantly higher expression of CYP1A1 and CYP3A9 in the upper region (P < 0.05). In contrast, mdr1a expression was significantly lower in the upper region (P < 0.05). In rats with normal and SIU tissues, a significant difference was observed only in CYP2D2 expression in the upper region of the small intestine (P < 0.05).


Intestinal and hepatic expression of cytochrome P450s and mdr1a in rats with indomethacin-induced small intestinal ulcers.

Kawauchi S, Nakamura T, Yasui H, Nishikawa C, Miki I, Inoue J, Horibe S, Hamaguchi T, Tanahashi T, Mizuno S - Int J Med Sci (2014)

Changes in the mRNA expression of CYPs and mdr1a in the rat small intestine with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. Total RNA was extracted from the upper, middle and lower small intestine. Expression levels of CYP and mdr1a mRNA were determined by real-time PCR, as described in the Materials and Methods. The data for mRNA expression for the non-SIUs (open triangles) and SIU groups (closed triangles) are expressed as the ratio of the mean value for each target gene in the upper region of the small intestine of the control group (open circles). Points represent the individual data, and bars represent the mean value for 5 to 6 rats per group. *P < 0.05, statistically significant.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4196121&req=5

Figure 4: Changes in the mRNA expression of CYPs and mdr1a in the rat small intestine with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. Total RNA was extracted from the upper, middle and lower small intestine. Expression levels of CYP and mdr1a mRNA were determined by real-time PCR, as described in the Materials and Methods. The data for mRNA expression for the non-SIUs (open triangles) and SIU groups (closed triangles) are expressed as the ratio of the mean value for each target gene in the upper region of the small intestine of the control group (open circles). Points represent the individual data, and bars represent the mean value for 5 to 6 rats per group. *P < 0.05, statistically significant.
Mentions: Figure 4 shows the effect of INM on the relative mRNA expression of CYPs and mdr1a in the upper, middle, and lower regions of the small intestine. In control rats, all CYP expression levels measured were significantly higher in the upper region than in the lower region (P < 0.05). An inverse trend was observed to expression of mdr1a, but it was not significant (P = 0.0503). In rats with non-SIU and SIU tissues, CYP and mdr1a expression were similar to those observed in control rats, with relatively high expression in the upper region. In particular, there was significantly higher expression of CYP1A1 and CYP3A9 in the upper region (P < 0.05). In contrast, mdr1a expression was significantly lower in the upper region (P < 0.05). In rats with normal and SIU tissues, a significant difference was observed only in CYP2D2 expression in the upper region of the small intestine (P < 0.05).

Bottom Line: Gene expression of the CYP family of enzymes and mdr1a was measured with quantitative real-time polymerase chain reaction (qPCR).Although previous studies have shown a direct effect of INM on CYP3A activity, we could not confirm any change in hepatic CY3A4 expression (major isoform of human CYP3A) in vitro.INM-induced SIUs had a subtle effect on intestinal CYP expression, but had an apparent action on hepatic CYP, which was influenced, at least in part, by the secondary inflammation.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Medical Pharmaceutics, Kobe Pharmaceutical University, 4-19-1 Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan; ; 2. Educational Center for Clinical Pharmacy, Kobe Pharmaceutical University, 4-19-1 Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan;

ABSTRACT

Background: Non-steroidal anti-inflammatory drugs induce the serious side effect of small intestinal ulcerations (SIUs), but little information is available regarding the consequences to drug metabolism and absorption.

Aim: We examined the existence of secondary hepatic inflammation in rats with indomethacin (INM)-induced SIUs and assessed its relationship to the cytochrome P450 (CYP) and P-glycoprotein (mdr1a), the major drug-metabolizing factors in the small intestine and the liver.

Methods: Gene expression of the CYP family of enzymes and mdr1a was measured with quantitative real-time polymerase chain reaction (qPCR). Vancomycin (VCM), a poorly absorbed drug, was administered intraduodenally to rats with SIUs.

Results: INM induced SIUs predominantly in the lower region of the small intestine with high expression of inflammatory markers. Liver dysfunction was also observed, which suggested a secondary inflammatory response in rats with SIUs. In the liver of rats with SIUs, the expression of CYP2C11, CYP2E1, and CYP3A1 was significantly decreased, and loss of CYP3A protein was observed. Although previous studies have shown a direct effect of INM on CYP3A activity, we could not confirm any change in hepatic CY3A4 expression (major isoform of human CYP3A) in vitro. The plasma VCM concentration was increased in rats with SIUs due to partial absorption from the mucosal injury, but not in normal mucosa.

Conclusions: INM-induced SIUs had a subtle effect on intestinal CYP expression, but had an apparent action on hepatic CYP, which was influenced, at least in part, by the secondary inflammation. Furthermore, drug absorption was increased in rats with SIUs.

Show MeSH
Related in: MedlinePlus