Effects of P-glycoprotein and Mrp2 inhibitors on the hepatobiliary disposition of Rhodamine 123 and its glucuronidated metabolite in isolated perfused rat livers

Parasrampuria, Ridhi; Mehvar, Reza

doi:10.1002/jps.21831

Cited by 20 publications

(21 citation statements)

References 31 publications

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“…The reduced biliary excretion rate found in our study may also be a consequence of posttranscriptional regulation or factors such as Pgp in vivo half-life (51,52). A recent study showed that the excretion of Rh123 was substantially reduced (90%) in the presence of the Pgp inhibitor cyclosporine A (53). Moreover, our group showed significantly increased fluorescence of Rh123 in hepatocytes after cyclosporine A treatment (30).…”

Section: Discussionsupporting

confidence: 56%

Effects of Long-Term Hepatic Ischemia-Reperfusion Injury on the Function of P-glycoprotein in vivo in Rats

Thorling¹,

Roberts²,

Liu³

et al. 2014

J Pharm Pharm Sci

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-PURPOSE: Ischemia-reperfusion injury is a common complication in liver surgery with oxidative stress related graft failure as a potential complication. The oxidative stress could affect hepatic drug transporters such as P-glycoprotein, which is crucial in the hepatic clearance of certain immunosuppressant drugs. Thus,, it is important to study its function after ischemia-reperfusion injury in vivo. Rhodamine 123 is a fluorescent substrate of P-glycoprotein and its hepatic disposition can be visualized using multiphoton microscopy in vivo using anaesthetized animals. The aim of this study was to investigate the effect of long-term ischemia-reperfusion injury on P-glycoprotein function in hepatocytes using in vivo multiphoton microscopy. METHODS: Localized ischemia was induced for 1 hour in rats. The liver was reperfused for 4, 24, 48 hours or 1 week, where-after rhodamine 123 was injected intravenously. Multiphoton microscopy imaged the liver and bile was collected continuously up to 6 hours following drug administration. The liver was harvested for histology andprotein expression of Pglycoprotein. RESULTS: Ischemia-reperfusion injury resulted in extensive liver damage, inflammatory cell infiltration and apoptosis in the midzonal and centrilobular regions of the liver acinus. P-glycoprotein protein expression decreased. Cellular concentration of rhodamine 123 increased as visualized by multiphoton microscopy, which was confirmed with decreased excretion of rhodamine 123 in collected bile. CONCLUSIONS: This study showed reduced function of P-glycoprotein in ischemia-reperfusion injury as reflected by decreased biliary excretion of Rhodamine 123, as well as reduced protein expression of the transporter. Multiphoton microscopy could be used to visualize and quantitate the intracellular levels of rhodamine 123. These findings stipulate the importance of using multiphoton microscopy to understand transmembrane drug flux and reflect on careful drug dosing after hepatic surgery.

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Section: Discussionsupporting

confidence: 56%

Effects of Long-Term Hepatic Ischemia-Reperfusion Injury on the Function of P-glycoprotein in vivo in Rats

Thorling¹,

Roberts²,

Liu³

et al. 2014

J Pharm Pharm Sci

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“…The ability of placental ABC transporters to hinder maternal-to-fetal and to accelerate fetal-to-maternal transport in a concentration-dependent manner has been demonstrated through perfusion of isolated rat placenta with rhodamine 123 and BODIPY-FL-prasozin [178]. Interestingly, the perfusion of organ models with fluorescent dyes can be performed in the absence or presence of transporter modulators, thus permitting to characterize the effects of transporter inhibitors at the organ level [179]. The consequences of induction of transporter expression by chemicals in terms of transport activity can additionally be studied [180].…”

Section: Analysis Of Drug Transporter Activity At the Organ Levelmentioning

confidence: 97%

Nature and uses of fluorescent dyes for drug transporter studies

Fardel

Vée²,

Jouan³

et al. 2015

Expert Opinion on Drug Metabolism & Toxicology

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A wide range of fluorescent dyes is now available for use in various aspects of drug transporter studies. The use of these dyes for transporter analyses may, however, be hampered by classic pitfalls of fluorescence technology, such as quenching. Transporter-independent processes such as passive diffusion of dyes through plasma membrane or dye sequestration into subcellular compartments must also be considered, as well as the redundant handling by various distinct transporters of some fluorescent probes. Finally, standardization of dye-based transport assays remains an important on-going issue.

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“…_________________________________________ The cationic dye rhodamine 123 (RH-123) is a P-gp substrate, which has been used extensively as a marker of P-gp activity in vitro in cells, ex vivo in isolated organs, and in vivo in whole animals. We have recently (12) characterized the hepatobiliary disposition of RH-123 in an isolated perfused liver (IPRL) model and showed that the biliary excretion of the dye is reduced after pharmacological inhibition of P-gp activity (13). Additionally, it was shown that the biliary clearance of the glucuronidated metabolite of the dye (RH-Glu) is sensitive to the modulation of Mrp2 activity (13).…”

Section: Introductionmentioning

confidence: 99%

“…We have recently (12) characterized the hepatobiliary disposition of RH-123 in an isolated perfused liver (IPRL) model and showed that the biliary excretion of the dye is reduced after pharmacological inhibition of P-gp activity (13). Additionally, it was shown that the biliary clearance of the glucuronidated metabolite of the dye (RH-Glu) is sensitive to the modulation of Mrp2 activity (13). Based on these observations, it was suggested that RH-123 may serve as a dual marker for the functions of both P-gp and Mrp2 in the IPRL model via simultaneous monitoring of the biliary clearances of the parent drug and the generated glucuronide metabolite, respectively (13).…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, it was shown that the biliary clearance of the glucuronidated metabolite of the dye (RH-Glu) is sensitive to the modulation of Mrp2 activity (13). Based on these observations, it was suggested that RH-123 may serve as a dual marker for the functions of both P-gp and Mrp2 in the IPRL model via simultaneous monitoring of the biliary clearances of the parent drug and the generated glucuronide metabolite, respectively (13). Therefore, we selected RH-123 as a marker for our functional studies in the present study.…”

Section: Introductionmentioning

confidence: 99%

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Effects of In Vivo Hepatic Ischemia-Reperfusion Injury on the Hepatobiliary Disposition of Rhodamine 123 and its Metabolites in Isolated Perfused Rat Livers

2012

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-Purpose.A few studies have shown that normothermic hepatic ischemia-reperfusion (IR) injury may affect the mRNA and/or protein levels of canalicular transporters P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (Mrp2). However, the effects of the injury on the functions of these canalicular transporters with respect to the biliary excretion of drugs remain largely unknown. Therefore, the purpose of this study was to investigate the effects of warm hepatic IR on the hepatobiliary disposition of rhodamine 123 (RH-123), a P-gp substrate, and its glucuronidated metabolite (RH-Glu), an Mrp2 substrate, in rats. Methods. Twenty four or 72 h following a 60-min partial ischemia or sham operation in rats, livers were isolated and perfused ex vivo with a constant concentration (~100 ng/mL) of RH-123. The concentration of RH-123 and its glucuronidated (RH-Glu) and deacylated (RH-110) metabolites were determined in the outlet perfusate, bile, and the liver tissue using HPLC, and relevant pharmacokinetic parameters were estimated. Results. Twentyfour-h IR caused a significant reduction in the hepatic extraction ratio of RH-123 (IR: 0.857 ± 0.078; Sham: 0.980 ± 0.017) and the biliary recovery of the parent drug and RH-Glu by 43% and 44%, respectively. The reductions in the biliary recovery were associated with significant reductions in the apparent biliary clearance of RH-123 and RH-Glu. Mass balance data showed that the formation of the glucuronidated or deacylated metabolite was not significantly affected by the 24-h IR injury. In contrast to the 24-h IR, the injury did not have any effect on the hepatobiliary disposition of RH-123 or its metabolites following 72 h of reperfusion. Conclusions. It is concluded that the pharmacokinetics of drugs that are subject to biliary excretion by the canalicular P-gp and Mrp2 transporters may be altered shortly after hepatic IR injury.

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Effects of P-glycoprotein and Mrp2 inhibitors on the hepatobiliary disposition of Rhodamine 123 and its glucuronidated metabolite in isolated perfused rat livers

Cited by 20 publications

References 31 publications

Effects of Long-Term Hepatic Ischemia-Reperfusion Injury on the Function of P-glycoprotein in vivo in Rats

Effects of Long-Term Hepatic Ischemia-Reperfusion Injury on the Function of P-glycoprotein in vivo in Rats

Nature and uses of fluorescent dyes for drug transporter studies

Effects of In Vivo Hepatic Ischemia-Reperfusion Injury on the Hepatobiliary Disposition of Rhodamine 123 and its Metabolites in Isolated Perfused Rat Livers

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