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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 mRNA expression of CYP and mdr1a and CYP3A protein in the rat liver with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. (A) Total RNA was extracted from the liver. 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 are expressed as the ratio of the mean value for CYP1A2 mRNA in the liver of the control group. Points represent individual data for the control group (open circles; N = 5) and SIU group (open squares; N = 6), and bars represent the mean value for each group. (B) The protein level of CYP3A was determined by western blot analysis, as described in the Materials and Methods. Densitometric quantification of CYP3A was performed and normalized to those of β-actin as a loading control. Data are presented as the mean values ± standard error for 4 rats per group. White columns represent those in the control group, and black columns represent those in the SIU group. *P < 0.05, statistically significant.
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Figure 5: Changes in mRNA expression of CYP and mdr1a and CYP3A protein in the rat liver with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. (A) Total RNA was extracted from the liver. 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 are expressed as the ratio of the mean value for CYP1A2 mRNA in the liver of the control group. Points represent individual data for the control group (open circles; N = 5) and SIU group (open squares; N = 6), and bars represent the mean value for each group. (B) The protein level of CYP3A was determined by western blot analysis, as described in the Materials and Methods. Densitometric quantification of CYP3A was performed and normalized to those of β-actin as a loading control. Data are presented as the mean values ± standard error for 4 rats per group. White columns represent those in the control group, and black columns represent those in the SIU group. *P < 0.05, statistically significant.

Mentions: Figure 5A shows the effect of INM on the relative expression of CYPs and mdr1a in the liver. The levels of CYP2C11, CYP2E1, and CYP3A1 mRNA expression were significantly decreased by more than 80% 24 h after the administration of INM (P < 0.05). The mRNA levels of CYP1A2, CYP2B1, CYP3A9, and mdr1a also decreased by 18.9%, 63.2%, 52.3%, and 34.4%, respectively, compared to control rats, but the differences were not significant.


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 mRNA expression of CYP and mdr1a and CYP3A protein in the rat liver with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. (A) Total RNA was extracted from the liver. 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 are expressed as the ratio of the mean value for CYP1A2 mRNA in the liver of the control group. Points represent individual data for the control group (open circles; N = 5) and SIU group (open squares; N = 6), and bars represent the mean value for each group. (B) The protein level of CYP3A was determined by western blot analysis, as described in the Materials and Methods. Densitometric quantification of CYP3A was performed and normalized to those of β-actin as a loading control. Data are presented as the mean values ± standard error for 4 rats per group. White columns represent those in the control group, and black columns represent those in the SIU group. *P < 0.05, statistically significant.
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Figure 5: Changes in mRNA expression of CYP and mdr1a and CYP3A protein in the rat liver with INM treatment. Animals were sacrificed 24 h after INM or vehicle administration. (A) Total RNA was extracted from the liver. 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 are expressed as the ratio of the mean value for CYP1A2 mRNA in the liver of the control group. Points represent individual data for the control group (open circles; N = 5) and SIU group (open squares; N = 6), and bars represent the mean value for each group. (B) The protein level of CYP3A was determined by western blot analysis, as described in the Materials and Methods. Densitometric quantification of CYP3A was performed and normalized to those of β-actin as a loading control. Data are presented as the mean values ± standard error for 4 rats per group. White columns represent those in the control group, and black columns represent those in the SIU group. *P < 0.05, statistically significant.
Mentions: Figure 5A shows the effect of INM on the relative expression of CYPs and mdr1a in the liver. The levels of CYP2C11, CYP2E1, and CYP3A1 mRNA expression were significantly decreased by more than 80% 24 h after the administration of INM (P < 0.05). The mRNA levels of CYP1A2, CYP2B1, CYP3A9, and mdr1a also decreased by 18.9%, 63.2%, 52.3%, and 34.4%, respectively, compared to control rats, but the differences were not significant.

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