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Effects of Hepatic Ischemia-Reperfusion Injury on the P-Glycoprotein Activity at the Liver Canalicular Membrane and Blood–Brain Barrier Determined by In Vivo Administration of Rhodamine 123 in Rats

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ABSTRACT

Purpose

To investigate the effects of normothermic hepatic ischemia-reperfusion (IR) injury on the activity of P-glycoprotein (P-gp) in the liver and at the blood–brain barrier (BBB) of rats using rhodamine 123 (RH-123) as an in vivo marker.

Methods

Rats were subjected to 90 min of partial ischemia or sham surgery, followed by 12 or 24 h of reperfusion. Following intravenous injection, the concentrations of RH-123 in blood, bile, brain, and liver were used for pharmacokinetic calculations. The protein levels of P-gp and some other transporters in the liver and brain were also determined by Western blot analysis.

Results

P-gp protein levels at the liver canalicular membrane were increased by twofold after 24 h of reperfusion. However, the biliary excretion of RH-123 was reduced in these rats by 26%, presumably due to IR-induced reductions in the liver uptake of the marker and hepatic ATP concentrations. At the BBB, a 24% overexpression of P-gp in the 24-h IR animals was associated with a 30% decrease in the apparent brain uptake clearance of RH-123. The pharmacokinetics or brain distribution of RH-123 was not affected by the 12-h IR injury.

Conclusions

Hepatic IR injury may alter the peripheral pharmacokinetics and brain distribution of drugs that are transported by P-gp and possibly other transporters.

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Abbreviations

ALT:

Alanine aminotransferase

AUC:

Area under the plasma concentration-time curve

BBB:

Blood–brain barrier

BIBF:

Bile salt-independent bile flow

C 40 br :

Concentration in the brain at 40 min

Cliver :

Concentration in the liver

Cmedian :

Concentration in the median lobe of the liver

Cright :

Concentration in the right lobe of the liver

DTT:

DL-dithiothreitol

f u :

Unbound fraction in plasma

IPRL:

Isolated perfused rat liver

IR:

Ischemia-reperfusion

K in :

Apparent brain uptake clearance

LPS:

Lipopolysaccharides

Mrp:

Multidrug resistance-associated protein

Oatp:

Organic anion transporting polypeptide

P-gp:

P-glycoprotein

PMSF:

Phenylmethanesulfonyl fluoride

RH-110:

Rhodamine 110

RH-123:

Rhodamine 123

RH-Glu:

Rhodamine glucuronide

TNF- α:

Tumor necrosis factor-α

UDPGA:

Uridine 5′-diphosphoglucuronic acid

Wischemic :

Weight of ischemic lobes of the liver

Wliver :

Total weight of the liver

Wnon-ischemic :

Weight of non-ischemic lobes of the liver

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ACKNOWLEDGMENTS AND DISCLOSURES

Mohammad K. Miah and Imam H. Shaik contributed equally to this work. The authors would like to acknowledge financial support from the Blood–brain Barrier Research Center at Texas Tech School of Pharmacy. Additionally, we would like to thank Dr. Thomas J. Thekkumkara for the use of VersaDoc Image System in his laboratory.

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Author notes

  1. Imam H. Shaik

    Present address: School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, Amarillo, Texas, USA

    Mohammad K. Miah, Imam H. Shaik, Ulrich Bickel & Reza Mehvar

  2. Center for Blood–Brain Barrier Research School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, Texas, USA

    Ulrich Bickel & Reza Mehvar

  3. School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter, Amarillo, Texas, 79106, USA

    Reza Mehvar

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  1. Mohammad K. Miah
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Correspondence to Reza Mehvar.

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Miah, M.K., Shaik, I.H., Bickel, U. et al. Effects of Hepatic Ischemia-Reperfusion Injury on the P-Glycoprotein Activity at the Liver Canalicular Membrane and Blood–Brain Barrier Determined by In Vivo Administration of Rhodamine 123 in Rats. Pharm Res 31, 861–873 (2014). https://doi.org/10.1007/s11095-013-1208-z

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