MilliporeSigma
  • Home
  • Search Results
  • Glucosamine-induced Sp1 O-GlcNAcylation ameliorates hypoxia-induced SGLT dysfunction in primary cultured renal proximal tubule cells.

Glucosamine-induced Sp1 O-GlcNAcylation ameliorates hypoxia-induced SGLT dysfunction in primary cultured renal proximal tubule cells.

Journal of cellular physiology (2014-03-05)
Han Na Suh, Yu Jin Lee, Mi Ok Kim, Jung Min Ryu, Ho Jae Han
ABSTRACT

The aim of this study is to determine whether GlcN could recover the endoplasmic reticulum (ER) stress-induced dysfunction of Na(+) /glucose cotransporter (SGLT) in renal proximal tubule cells (PTCs) under hypoxia. With the rabbit model, the renal ischemia induced tubulointerstitial abnormalities and decreased SGLTs expression in tubular brush-border, which were recovered by GlcN. Thus, the protective mechanism of GlcN against renal ischemia was being examined by using PTCs. Hypoxia decreased the level of protein O-GlcNAc and the expression of O-GlcNAc transferase (OGT) while increased O-GlcNAcase (OGA) and these were reversed by GlcN. Hypoxia also decreased the expression of SGLTs (SGLT1 and 2) and [(14) C]-α-methyl-D-glucopyranoside (α-MG) uptake which were recovered by GlcN and PUGNAc (OGA inhibitor). Hypoxia enhanced reactive oxygen species (ROS) and then ER stress proteins, glucose-regulated protein 78 (GRP78), and C/EBP-homologous protein (CHOP). However, the expression of GRP78 increased till 6 h and then decreased whereas CHOP increased gradually. Moreover, decreased GRP78 and increased CHOP were reversed by NAC (antioxidant) and GlcN. GlcN ameliorated hypoxia-induced decrease of O-GlcNAc modification of Sp1 but OGT or Sp1 siRNAs blocked the recovery effect of GlcN on SGLT expression and α-MG uptake. In addition, hypoxia-decreased GRP78 and HIF-1α expression was reversed by GlcN but OGT siRNA or Sp1 siRNA ameliorated the effect of GlcN. When PTCs were transfected with GRP78 siRNA or HIF-1α siRNA, SGLT expression and α-MG uptake was decreased. Taken together, these data suggest that GlcN-induced O-GlcNAc modified Sp1 with stimulating GRP78 and HIF-1α activity ameliorate hypoxia-induced SGLT dysfunction in renal PTCs. J. Cell. Physiol. 229: 1557-1568, 2014. © 2014 Wiley Periodicals, Inc.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Magnesium sulfate, puriss. p.a., drying agent, anhydrous, ≥98.0% (KT), powder (very fine)
Sigma-Aldrich
Magnesium sulfate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Magnesium sulfate, anhydrous, free-flowing, Redi-Dri, reagent grade, ≥97%
Sigma-Aldrich
Magnesium sulfate, ≥99.99% trace metals basis
Sigma-Aldrich
MISSION® esiRNA, targeting mouse Hspa5
Sigma-Aldrich
N-Acetyl-L-cysteine, BioReagent, suitable for cell culture
Sigma-Aldrich
Fluorescein 5(6)-isothiocyanate, ≥90% (HPLC)
Sigma-Aldrich
N-Acetyl-L-cysteine, Sigma Grade, ≥99% (TLC), powder
Sigma-Aldrich
N-Acetyl-L-cysteine, BioXtra, ≥99% (TLC)
Sigma-Aldrich
Phosphorylase b from rabbit muscle, lyophilized powder, ≥20 units/mg protein
Sigma-Aldrich
Magnesium sulfate, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Phosphorylase b from rabbit muscle, For use as a marker in SDS-PAGE
Sigma-Aldrich
Magnesium sulfate, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Propidium iodide solution, solution (1.0 mg/ml in water)
Sigma-Aldrich
Fluorescein 5(6)-isothiocyanate, BioReagent, suitable for fluorescence, mixture of 2 components, ≥90% (HPLC)
Glucosamine hydrochloride, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Magnesium sulfate, anhydrous, reagent grade, ≥97%
Supelco
N-Acetyl-L-cysteine, Pharmaceutical Secondary Standard; Certified Reference Material
Glucosamine for system suitability, European Pharmacopoeia (EP) Reference Standard
USP
Glucosamine hydrochloride, United States Pharmacopeia (USP) Reference Standard
Supelco
Glucosamine hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
N-Acetylglucosamine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Magnesium sulfate solution, for molecular biology, 1.00 M±0.04 M
Sigma-Aldrich
Sodium chloride, 99.999% trace metals basis
Sigma-Aldrich
Sodium chloride solution, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Sodium chloride solution, 5 M
Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
Sigma-Aldrich
Fluorescein isothiocyanate isomer I, ≥97.5% (HPLC)
Sigma-Aldrich
Fluorescein isothiocyanate isomer I, suitable for protein labeling, ≥90% (HPLC), powder