Decreased Expression of Multidrug Resistance-Associated Protein 4 (MRP4/ABCC4) Leads to Reduced Glucuronidation of Flavonoids in UGT1A1-Overexpressing HeLa Cells: The Role of Futile Recycling

Sun, Hua; Zhou, Xiaotong; Zhang, Qizhou; Wu, Baojian

doi:10.1021/acs.jafc.5b00983

Cited by 21 publications

(30 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moreover, the first-pass metabolism by phase II enzymes limits the oral bioavailability of many drugs and active herbal-derived chemicals (Wu et al, 2011). The present study and our previous one (Sun et al, 2015b) indicated that cellular conjugation is regulated by efflux transporters. Hence, the inhibition of both enzymes and transporters represented a novel and promising strategy to enhance the oral bioavailability of drugs undergoing extensive conjugation, although the inhibition of enzyme activity alone showed potential in enhancing resveratrol bioavailability (Zhou et al, 2015).…”

supporting

confidence: 63%

“…Impaired sulfated metabolite excretion resulted in its accumulation, thereby enhancing desulfonation and reducing total sulfonation. The current study and our previous one highlighted that the deconjugation (or futile recycling) process was necessary for the interplay of phase II enzymes with efflux transporters to occur (Sun et al, 2015b). …”

Section: Discussionmentioning

confidence: 50%

“…The dependence of phase II metabolism on efflux transporters is underpinned by the fact that the hydrophilic phase II metabolites (lacking in passive transport ability) require the efflux transporters for cellular excretion (Wu, 2012). The futile recycling (i.e., b-glucuronidase-mediated deglucuronidation) has been suggested to be the main mechanism underlying the glucuronidation-transport interplay (Sun et al, 2015b). In contrast to the well-established interplay of glucuronidation with efflux transport, little is known about whether and how the sulfonation-transport interplay occurs.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3

Zhao¹,

Wang²,

Li³

et al. 2016

Drug Metabolism and Disposition

View full text Add to dashboard Cite

Elucidating the intricate relationships between metabolic and transport pathways contributes to improved predictions of in vivo drug disposition and drug-drug interactions. Here we reported that inhibited excretion of conjugative metabolites [i.e., hesperetin 39-O-sulfate (H39S) and hesperetin 7-O-sulfate (H7S)] by MK-571 led to reduced metabolism of hesperetin (a maximal 78% reduction) in human embryonic kidney 293 cells overexpressing sulfotransferase 1A3 (named SULT293 cells). The strong dependence of cellular sulfonation on the efflux transport of generated sulfated metabolites revealed an interplay of sulfonation metabolism with efflux transport (or sulfonation-transport interplay). Polymerase chain reaction (PCR) and Western blot analyses demonstrated that SULT293 cells expressed multiple sulfatases such as arylsulfatase A (ARSA), ARSB, and ARSC. Of these three desulfonation enzymes, only ARSB showed significant activities toward hesperetin sulfates. The intrinsic clearance values for the hydrolysis of H39S and H7S were estimated at 0.6 and 0.5 ml/h/mg, respectively. Furthermore, knockdown of ARSB attenuated the regulatory effect of efflux transporter on cellular sulfonation, whereas overexpression of ABSB enhanced the transporter effect. Taken together, the results indicated that ARSB mediated the sulfonation-transport interplay in SULT293 cells.

show abstract

supporting

confidence: 63%

Section: Discussionmentioning

confidence: 50%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3

Zhao¹,

Wang²,

Li³

et al. 2016

Drug Metabolism and Disposition

View full text Add to dashboard Cite

show abstract

“…A different set of experiments was performed to obtain the hesperetin/sulfates levels (both extracellular and intracellular) versus time profiles for pharmacokinetic modeling as described (Sun et al, 2015). In brief, the cells were incubated with hesperetin at a dose of 5 nmol.…”

Section: Sulfate Excretion Experimentsmentioning

confidence: 99%

Multidrug Resistance-Associated Protein 4 (MRP4/ABCC4) Controls Efflux Transport of Hesperetin Sulfates in Sulfotransferase 1A3–Overexpressing Human Embryonic Kidney 293 Cells

et al. 2015

Self Cite

View full text Add to dashboard Cite

Sulfonation is an important metabolic pathway for hesperetin. However, the mechanisms for the cellular disposition of hesperetin and its sulfate metabolites are not fully established. In this study, disposition of hesperetin via the sulfonation pathway was investigated using human embryonic kidney (HEK) 293 cells overexpressing sulfotransferase 1A3. Two monosulfates, hesperetin-39-O-sulfate (H-39-S) and hesperetin-7-O-sulfate (H-7-S), were rapidly generated and excreted into the extracellular compartment upon incubation of the cells with hesperetin. Regiospecific sulfonation of hesperetin by the cell lysate followed the substrate inhibition kinetics (V max = 0.66 nmol/min per mg, K m = 12.9 mM, and K si = 58.1 mM for H-39-S; V max = 0.29 nmol/min per mg, K m = 14.8 mM, and K si = 49.1 mM for H-7-S). The pan-multidrug resistance-associated protein (MRP) inhibitor MK-571 at 20 mM essentially abolished cellular excretion of both H-39-S and H-7-S (the excretion activities were only 6% of the control), whereas the breast cancer resistance protein-selective inhibitor Ko143 had no effects on sulfate excretion. In addition, knockdown of MRP4 led to a substantial reduction (>47.1%; P < 0.01) in sulfate excretion. Further, H-39-S and H-7-S were good substrates for transport by MRP4 according to the vesicular transport assay. Moreover, sulfonation of hesperetin and excretion of its metabolites were well characterized by a two-compartment pharmacokinetic model that integrated drug uptake and sulfonation with MRP4-mediated sulfate excretion. In conclusion, the exporter MRP4 controlled efflux transport of hesperetin sulfates in HEK293 cells. Due to significant expression in various organs/tissues (including the liver and kidney), MRP4 should be a determining factor for the elimination and body distribution of hesperetin sulfates.

show abstract

“…Establishment of UGT1A1-and GUSB-overexpressing HeLa Cells. UGT1A1-or GUSB-overexpressing HeLa cells were generated using the lentiviral transfection method as previously described (Quan et al, 2015;Sun et al, 2015). Western blotting against GUSB was performed as previously described (Quan et al, 2015).…”

Section: Methodsmentioning

confidence: 99%

Efflux Transport Characterization of Resveratrol Glucuronides in UDP-Glucuronosyltransferase 1A1 Transfected HeLa Cells: Application of a Cellular Pharmacokinetic Model to Decipher the Contribution of Multidrug Resistance-Associated Protein 4

Wang

Feng

Quan

et al. 2016

Drug Metabolism and Disposition

View full text Add to dashboard Cite

Resveratrol undergoes extensive metabolism to form biologically active glucuronides in humans. However, the transport mechanisms for resveratrol glucuronides are not fully established. Here, we aimed to characterize the efflux transport of resveratrol glucuronides using UGT1A1-overexpressing HeLa cells (HeLa1A1 cells), and to determine the contribution of multidrug resistanceassociated protein (MRP) 4 to cellular excretion of the glucuronides. Two glucuronide isomers [i.e., resveratrol 3-O-glucuronide (R3G) and resveratrol 49-O-glucuronide (R49G)] were excreted into the extracellular compartment after incubation of resveratrol (1-100 mM) with HeLa1A1 cells. The excretion rate was linearly related to the level of intracellular glucuronide, indicating that glucuronide efflux was a nonsaturable process. MK-571 (a dual inhibitor of UGT1A1 and MRPs) significantly decreased the excretion rates of R3G and R49G while increasing their intracellular levels. Likewise, short-hairpin RNA (shRNA)-mediated silencing of MRP4 caused a significant reduction in glucuronide excretion but an elevation in glucuronide accumulation. Furthermore, b-glucuronidase expressed in the cells catalyzed the hydrolysis of the glucuronides back to the parent compound. A cellular pharmacokinetic model integrating resveratrol transport/metabolism with glucuronide hydrolysis/excretion was well fitted to the experimental data, allowing derivation of the efflux rate constant values in the absence or presence of shRNA targeting MRP4. It was found that a large percentage of glucuronide excretion (43%-46%) was attributed to MRP4. In conclusion, MRP4 participated in cellular excretion of R3G and R49G. Integration of mechanistic pharmacokinetic modeling with transporter knockdown was a useful method to derive the contribution percentage of an exporter to overall glucuronide excretion.

show abstract

Decreased Expression of Multidrug Resistance-Associated Protein 4 (MRP4/ABCC4) Leads to Reduced Glucuronidation of Flavonoids in UGT1A1-Overexpressing HeLa Cells: The Role of Futile Recycling

Cited by 21 publications

References 42 publications

Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3

Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3

Multidrug Resistance-Associated Protein 4 (MRP4/ABCC4) Controls Efflux Transport of Hesperetin Sulfates in Sulfotransferase 1A3–Overexpressing Human Embryonic Kidney 293 Cells

Efflux Transport Characterization of Resveratrol Glucuronides in UDP-Glucuronosyltransferase 1A1 Transfected HeLa Cells: Application of a Cellular Pharmacokinetic Model to Decipher the Contribution of Multidrug Resistance-Associated Protein 4

Contact Info

Product

Resources

About