Ursodeoxycholate Restores Biliary Excretion of Methotrexate in Rats with Ethinyl Estradiol Induced-Cholestasis by Restoring Canalicular Mrp2 Expression

Abstract

The in vivo relevance of ursodeoxycholate (UDCA) treatment (100 mg/kg/day, per oral tid for 5 days before cholestasis induction followed by the same dosing for 5 days) on hepatic function was investigated in rats with 17α-ethinylestradiol (EE, 10 mg/kg, subcutaneous for 5 days)-induced experimental cholestasis. The bile flow rate and the expression level of hepatic multidrug resistance-associated protein 2 (Mrp 2) that were decreased in cholestasis were restored after UDCA treatment. Consistent with this, the biliary excretion clearance (CLexc,bile) of a representative Mrp2 substrate—methotrexate (MTX)—was decreased in cholestatic rats but was restored after UDCA treatment. Consequently, the plasma concentrations of MTX, which were increased by cholestasis, were decreased to control levels by UDCA treatment. Thus, the restoration of CLexc,bile appears to be associated with the increase in Mrp2 expression on the canalicular membrane by UDCA treatment followed by Mrp2-mediated biliary excretion of MTX. On the other hand, the hepatic uptake clearance (CLup,liver) of MTX was unchanged by cholestasis or UDCA treatment, suggestive of the absence of any association between the uptake process and the overall biliary excretion of MTX. Since UDCA has been known to induce the expression of canalicular MRP2 in humans, UDCA treatment might be effective in humans to maintain or accelerate the hepatobiliary elimination of xenobiotics or metabolic conjugates that are MRP2 substrates.

Effects of Red Ginseng Extract on the Pharmacokinetics and Elimination of Methotrexate via Mrp2 Regulation

“…CL bile accounted for 70% of the CL total of the intravenous bolus injection of methotrexate at a dose of 3 mg/kg in the control group and CL urine accounted for a relatively minor portion (18%, Table 4 ) in this group. These results showed that biliary excretion is the major pathway of methotrexate elimination, which is consistent with the findings of a previous study [ 21 ] and Mrp2 could be a controlling factor in the pharmacokinetics of methotrexate [ 18 ]. Therefore, the decrease in the expression of Mrp2 might contribute to the decreased biliary excretion of methotrexate and consequently a 1.5-fold increase in plasma methotrexate exposure ( Table 4 ).…”

“…RT-PCR of Mrp1, Mrp2, Bsep, and P-gp was performed using a LightCycler 96 real-time PCR system (Roche, Carlsbad, CA, USA) as previously described [ 21 , 37 ]. Primers were designed using ProbeFinder as follows: 5′-tgagggtggagaaaaggttg-5′ and 5′-aaacccagggtgagagatga-3′ for Mrp1 (NM_022281.2); 5′-aatacatgaccttttggtgtttctg-5′ and 5′-acgaaaccgatcagcaactt-3′ for Mrp2 (NM_012833.2); 5′-gggcagtcacacccatctac-5′ and 5′-ctttatcgaggagtgaaaaagtcc-3′ for Bsep (NM_031760.1); 5′-cacagaccgtcagcgaca-5′ and 5′-caatgcccgtgtaatagtaggc-3′ for P-gp (NM_012623.2).…”

“…For example, numerous anionic drugs such as methotrexate, SN38, cisplatin, vinblastine, and sulfinpyrazone are mainly eliminated from the body by Mrp2 [ 19 , 20 ]. GSH facilitates the biliary excretion of amphipathic drugs via Mrp2 through the conjugation reaction and also acts as a regulator of redox homeostasis [ 21 ]. RGE has been shown to be effective in reducing oxidative stress through the Nrf2 signaling pathway [ 7 , 22 ].…”

“…RGE has been shown to be effective in reducing oxidative stress through the Nrf2 signaling pathway [ 7 , 22 ]. Nrf2 is a transcriptional factor that regulates Mrp2 expression [ 21 ], suggesting herb–drug interaction between red ginseng and Mrp2 substrates.…”

Prophylactic Fucose can Alleviate Lipopolysaccharide-Induced Cholestatic Liver Injury in Neonatal Rats

“…Bile acid excretion is mainly mediated by ABCB11 and MRP-2, upregulation, which are important adaptive mechanisms to prevent bile acid accumulation within hepatocytes in case of neonatal cholestasis. [12][13][14] In hepatocytes, ABCB-11 is localized within the apical membrane and deficiency of canalicular ABCB-11 function is associated with severe forms of cholestasis. 15 One study suggests that L-fucose is pharmacologically active at the dose of 100 mg/kg for investigating learning behaviors in rats, 16 as a human milk monosaccharide it seems safe.…”