Presence or absence of microbiome modulates the response of mice organism to administered drug nabumetone

Abstract

The gut microbiota provides a wide range of beneficial functions for the host, and has an immense effect on the host's health status. The presence of microbiome in the gut may often influence the effect of an orally administered drug. Molecular mechanisms of this process are however mostly unclear. We investigated how the effect of a nonsteroidal drug nabumetone on expression of drug metabolizing enzymes (DMEs) in mice intestine and liver is changed by the presence of microbiota, here, using the germ free (GF) and specific pathogen free (SPF) BALB/c mice. First, we have found in a preliminary experiment that in the GF mice there is a tendency to increase bioavailability of the active form of nabumetone, which we have found now to be possibly influenced by differences in expression of DMEs in the GF and SPF mice. Indeed, we have observed that the expression of the most of selected cytochromes P450 (CYPs) was significantly changed in the small intestine of GF mice compared to the SPF ones. Moreover, orally administered nabumetone itself altered the expression of some CYPs and above all, in different ways in the GF and SPF mice. In the GF mice, the expression of the DMEs (CYP1A) responsible for the formation of active form of the drug are significantly increased in the small intestine and liver after nabumetone application. These results highlight the importance of gut microbiome in processes involved in drug metabolism in the both gastrointestinal tract and in the liver with possible clinical relevance.

Fig. 1

Influence of nabumetone on mRNA expression of CYPs (A) and transcription factors (B) in small intestine of SPF and GF mice in first three hours after one dose application. Data are expressed as percentage of the respective SPF/GF untreated control. mRNA data represent the mean ± SD. from 9 individual animals. Values significantly increased or decreased (*P<0.05, **P<0.01, ***P<0.001) in comparison with the control are labeled. NABU – nabumetone.

Fig. 2

The protein expression of CYP1A (A), CYP2B1/2 (B), CYP3A (C) in the small intestine of SPF and GF mice treated with nabumetone in time after one dose application in comparison with their SPF/GF control group (100 %) without nabumetone treatment. Protein relative levels were measured in the pooled microsomal fractions (3 animals per group). NABU – nabumetone.

Fig. 3

The enzyme activity of CYP1A (A) and CYP2B (B) in the small intestine of SPF and GF mice treated with nabumetone in time after one dose application in comparison with their SPF/GF control group (100 %) without nabumetone treatment. Protein relative levels were measured in the pooled microsomal fraction (3 animals per group). NABU – nabumetone.

Fig. 4

Influence of nabumetone on mRNA expression of CYPs (A) and transcription factors (B) in liver of SPF and GF mice in first three hours after one dose application (1; 2 and 3 hours). Data are expressed as percentage of the SPF/GF control group (100 %) without nabumetone treatment. The mRNA data represent the mean ± SD from 9 individual animals. Values significantly increased or decreased (*P<0.05; **P<0.01; ***P<0.001) in comparison with the control are labeled. NABU – nabumetone.

Fig. 5

The protein expression of CYP1A (A) and CYP3A (B) in the liver of SPF and GF mice treated with nabumetone in time after one dose application in comparison with their SPF/GF control group (100 %) without nabumetone treatment. Protein relative levels were measured in the pooled microsomal fractions (3 animals per group).