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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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Area of SIUs (A), and intestinal iNOS mRNA expression (B) in rats with INM-induced SIUs. Animals were sacrificed 24 h after INM or vehicle administration. (A) Areas of SIU were microscopically assessed in control (open columns) and INM-treated rats (closed columns). The data are presented as the mean ± standard error for 5 to 6 rats per group. (B) Total RNA was extracted from the small intestine. The expression level of iNOS mRNA was 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 iNOS mRNA in the upper intestine of the control group. Points represent individual data for control (open circles), non-SIU (open triangles), and SIU (closed triangles) groups, and bars represent the mean value for 6 to 11 rats per group. *P < 0.05, statistically significant.
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Figure 1: Area of SIUs (A), and intestinal iNOS mRNA expression (B) in rats with INM-induced SIUs. Animals were sacrificed 24 h after INM or vehicle administration. (A) Areas of SIU were microscopically assessed in control (open columns) and INM-treated rats (closed columns). The data are presented as the mean ± standard error for 5 to 6 rats per group. (B) Total RNA was extracted from the small intestine. The expression level of iNOS mRNA was 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 iNOS mRNA in the upper intestine of the control group. Points represent individual data for control (open circles), non-SIU (open triangles), and SIU (closed triangles) groups, and bars represent the mean value for 6 to 11 rats per group. *P < 0.05, statistically significant.

Mentions: Animals treated with INM for 24 h exhibited ulcers throughout the small intestine, but animals not treated with INM (control group) had few ulcers (Fig. 1A). The average area of SIUs in the lower small intestine was 77.7 ± 16.6 mm2, which was significantly higher than those in the upper and middle sections of the small intestine (22.2 ± 5.1 and 18.5 ± 3.7 mm2, respectively) (P < 0.05). In rats with INM-induced SIUs, the level of expression of inducible nitric oxide synthase (iNOS) mRNA in the lower region of the small intestine was significantly higher than that in the upper region. In addition, there was a significant difference in iNOS mRNA expression levels between SIU and non-SIU tissue with INM treatment in the lower region of the small intestine (Fig. 1B).


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)

Area of SIUs (A), and intestinal iNOS mRNA expression (B) in rats with INM-induced SIUs. Animals were sacrificed 24 h after INM or vehicle administration. (A) Areas of SIU were microscopically assessed in control (open columns) and INM-treated rats (closed columns). The data are presented as the mean ± standard error for 5 to 6 rats per group. (B) Total RNA was extracted from the small intestine. The expression level of iNOS mRNA was 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 iNOS mRNA in the upper intestine of the control group. Points represent individual data for control (open circles), non-SIU (open triangles), and SIU (closed triangles) groups, and bars represent the mean value for 6 to 11 rats per group. *P < 0.05, statistically significant.
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Related In: Results  -  Collection

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

Figure 1: Area of SIUs (A), and intestinal iNOS mRNA expression (B) in rats with INM-induced SIUs. Animals were sacrificed 24 h after INM or vehicle administration. (A) Areas of SIU were microscopically assessed in control (open columns) and INM-treated rats (closed columns). The data are presented as the mean ± standard error for 5 to 6 rats per group. (B) Total RNA was extracted from the small intestine. The expression level of iNOS mRNA was 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 iNOS mRNA in the upper intestine of the control group. Points represent individual data for control (open circles), non-SIU (open triangles), and SIU (closed triangles) groups, and bars represent the mean value for 6 to 11 rats per group. *P < 0.05, statistically significant.
Mentions: Animals treated with INM for 24 h exhibited ulcers throughout the small intestine, but animals not treated with INM (control group) had few ulcers (Fig. 1A). The average area of SIUs in the lower small intestine was 77.7 ± 16.6 mm2, which was significantly higher than those in the upper and middle sections of the small intestine (22.2 ± 5.1 and 18.5 ± 3.7 mm2, respectively) (P < 0.05). In rats with INM-induced SIUs, the level of expression of inducible nitric oxide synthase (iNOS) mRNA in the lower region of the small intestine was significantly higher than that in the upper region. In addition, there was a significant difference in iNOS mRNA expression levels between SIU and non-SIU tissue with INM treatment in the lower region of the small intestine (Fig. 1B).

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