Abstract
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 3′-O-sulfate (H3′S) 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 H3′S and H7S were estimated at 0.6 and 0.5 μl/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.
Footnotes
- Received April 10, 2016.
- Accepted June 17, 2016.
M.Z. and S.W. contributed equally to this work.
This work was supported by the National Natural Science Foundation of China [Grant 81573488], the Young Scientist Special Projects in biotechnological and pharmaceutical field of 863 Program [Grant 2015AA020916], the Outstanding Youth Fund from the Natural Science Foundation of Guangdong Province [Grant 2014A030306014], and The PhD Start-up Fund of Natural Science Foundation of Guangdong Province [Grant 2015A030310339].
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- Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics
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