Supplementary Material for: Role of Organic Anion Transporters in the Uptake of Protein-Bound Uremic Toxins by Human Endothelial Cells and Monocyte Chemoattractant Protein-1 Expression
Favretto G.
Souza L.M.
Gregório P.C.
Cunha R.S.
Maciel R.A.P.
Sassaki G.L.
Toledo M.G.
Pecoits-Filho R.
Souza W.M.
Stinghen A.E.M.
10.6084/m9.figshare.4970480.v1
https://karger.figshare.com/articles/journal_contribution/Supplementary_Material_for_Role_of_Organic_Anion_Transporters_in_the_Uptake_of_Protein-Bound_Uremic_Toxins_by_Human_Endothelial_Cells_and_Monocyte_Chemoattractant_Protein-1_Expression/4970480
<p>Organic anion transporters (OATs) are involved in the uptake of
uremic toxins such as p-cresyl sulfate (PCS) and indoxyl sulfate (IS),
which play a role in endothelial dysfunction in patients with chronic
kidney diseases (CKD). In this study, we investigated the role of OAT1
and OAT3 in the uptake of PCS and IS into human endothelial cells. PCS
was synthesized via <i>p</i>-cresol sulfation and characterized using
analytical methods. The cells were treated with PCS and IS in the
absence and presence of probenecid (Pb), an OAT inhibitor. Cell
viability was assessed using the MTT assay. The absorbed toxins were
analyzed using chromatography, OAT expression using immunocytochemistry
and western blot, and monocyte chemoattractant protein-1 (MCP-1)
expression using enzyme-linked immunosorbent assay. Cell viability
decreased after toxin treatment in a dose-dependent manner. PCS and IS
showed significant internalization after 60 min treatment, while no
internalization was observed in the presence of Pb, suggesting that OATs
are involved in the transport of both toxins. Immunocytochemistry and
western blot demonstrated OAT1 and OAT3 expression in endothelial cells.
MCP-1 expression increased after toxins treatment but decreased after
Pb treatment. PCS and IS uptake were mediated by OATs, and OAT blockage
could serve as a therapeutic strategy to inhibit MCP-1 expression.</p>
2017-05-04 12:25:11
Chronic Kidney Disease Interplay
Endothelial dysfunction
Uremic toxins
Organic anion transporters
Monocyte chemoattractant protein-1