Short-term high-glucose treatment decreased abundance of Orai1 protein through posttranslational mechanisms in rat Mesangial cells

Hui Jiang, Shubiao Zou, Sarika Chaudhari, Rong Ma

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The short-term effect of high-glucose (HG) treatment on store-operated Ca 2+ entry in mesangial cells (MCs) is not well-known. The aim of the present study was to determine whether and how HG treatment for a short period altered protein abundance of Orai1, the channel mediating store-operated Ca 2+ entry in MCs. Rat and human MCs were exposed to HG (25 mM) for 2, 4, 8, and 24 h, and the abundance of Orai1 protein was significantly decreased at the time points of 8 and 16 h. Consistently, HG treatment for 8 h significantly reduced store-operated Ca 2+ entry in rat MCs. However, HG treatment for the same time periods did not alter the levels of Orai1 transcript. Cycloheximide, a protein synthesis inhibitor, did not affect the HG-induced decrease of Orai1 protein, suggesting a posttranslational mechanism was involved. However, the HG effect on Orai1 protein was significantly attenuated by MG132 (a ubiquitin-proteasome inhibitor) and NH 4 Cl (a lysosomal pathway inhibitor). Furthermore, HG treatment for 8 h stimulated ubiquitination of Orai1 protein. We further found that polyethylene glycol-catalase, an antioxidant, significantly blunted the HG-induced reduction of Orai1 protein. In support of involvement of reactive oxygen species in the HG effects, hydrogen peroxide (H 2 O 2 ) itself significantly decreased abundance of Orai1 protein and increased the level of ubiquitinated Orai1. Taken together, these results suggest that a short-term HG treatment decreased abundance of Orai1 protein in MCs by promoting the protein degradation through the ubiquitination-proteasome and –lysosome mechanisms. This HG-stimulated posttranslational mechanism was mediated by H 2 O 2 .

Original languageEnglish
Pages (from-to)F855-F863
JournalAmerican Journal of Physiology - Renal Physiology
Volume314
Issue number5
DOIs
StatePublished - 1 Jan 2018

Fingerprint

Mesangial Cells
Glucose
Proteins
Ubiquitination
Period Circadian Proteins
Proteasome Inhibitors
Protein Synthesis Inhibitors
Proteasome Endopeptidase Complex
Cycloheximide
Ubiquitin
Lysosomes
Hydrogen Peroxide
Proteolysis
Reactive Oxygen Species
Antioxidants

Keywords

  • Hydrogen peroxide
  • Mesangial cells
  • Orai1
  • Store-operated Ca entry
  • Ubiquitination

Cite this

@article{51160d66446e4a728c79c5e930d1b198,
title = "Short-term high-glucose treatment decreased abundance of Orai1 protein through posttranslational mechanisms in rat Mesangial cells",
abstract = "The short-term effect of high-glucose (HG) treatment on store-operated Ca 2+ entry in mesangial cells (MCs) is not well-known. The aim of the present study was to determine whether and how HG treatment for a short period altered protein abundance of Orai1, the channel mediating store-operated Ca 2+ entry in MCs. Rat and human MCs were exposed to HG (25 mM) for 2, 4, 8, and 24 h, and the abundance of Orai1 protein was significantly decreased at the time points of 8 and 16 h. Consistently, HG treatment for 8 h significantly reduced store-operated Ca 2+ entry in rat MCs. However, HG treatment for the same time periods did not alter the levels of Orai1 transcript. Cycloheximide, a protein synthesis inhibitor, did not affect the HG-induced decrease of Orai1 protein, suggesting a posttranslational mechanism was involved. However, the HG effect on Orai1 protein was significantly attenuated by MG132 (a ubiquitin-proteasome inhibitor) and NH 4 Cl (a lysosomal pathway inhibitor). Furthermore, HG treatment for 8 h stimulated ubiquitination of Orai1 protein. We further found that polyethylene glycol-catalase, an antioxidant, significantly blunted the HG-induced reduction of Orai1 protein. In support of involvement of reactive oxygen species in the HG effects, hydrogen peroxide (H 2 O 2 ) itself significantly decreased abundance of Orai1 protein and increased the level of ubiquitinated Orai1. Taken together, these results suggest that a short-term HG treatment decreased abundance of Orai1 protein in MCs by promoting the protein degradation through the ubiquitination-proteasome and –lysosome mechanisms. This HG-stimulated posttranslational mechanism was mediated by H 2 O 2 .",
keywords = "Hydrogen peroxide, Mesangial cells, Orai1, Store-operated Ca entry, Ubiquitination",
author = "Hui Jiang and Shubiao Zou and Sarika Chaudhari and Rong Ma",
year = "2018",
month = "1",
day = "1",
doi = "10.1152/ajprenal.00513.2017",
language = "English",
volume = "314",
pages = "F855--F863",
journal = "American Journal of Physiology - Renal Physiology",
issn = "0363-6127",
publisher = "American Physiological Society",
number = "5",

}

Short-term high-glucose treatment decreased abundance of Orai1 protein through posttranslational mechanisms in rat Mesangial cells. / Jiang, Hui; Zou, Shubiao; Chaudhari, Sarika; Ma, Rong.

In: American Journal of Physiology - Renal Physiology, Vol. 314, No. 5, 01.01.2018, p. F855-F863.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Short-term high-glucose treatment decreased abundance of Orai1 protein through posttranslational mechanisms in rat Mesangial cells

AU - Jiang, Hui

AU - Zou, Shubiao

AU - Chaudhari, Sarika

AU - Ma, Rong

PY - 2018/1/1

Y1 - 2018/1/1

N2 - The short-term effect of high-glucose (HG) treatment on store-operated Ca 2+ entry in mesangial cells (MCs) is not well-known. The aim of the present study was to determine whether and how HG treatment for a short period altered protein abundance of Orai1, the channel mediating store-operated Ca 2+ entry in MCs. Rat and human MCs were exposed to HG (25 mM) for 2, 4, 8, and 24 h, and the abundance of Orai1 protein was significantly decreased at the time points of 8 and 16 h. Consistently, HG treatment for 8 h significantly reduced store-operated Ca 2+ entry in rat MCs. However, HG treatment for the same time periods did not alter the levels of Orai1 transcript. Cycloheximide, a protein synthesis inhibitor, did not affect the HG-induced decrease of Orai1 protein, suggesting a posttranslational mechanism was involved. However, the HG effect on Orai1 protein was significantly attenuated by MG132 (a ubiquitin-proteasome inhibitor) and NH 4 Cl (a lysosomal pathway inhibitor). Furthermore, HG treatment for 8 h stimulated ubiquitination of Orai1 protein. We further found that polyethylene glycol-catalase, an antioxidant, significantly blunted the HG-induced reduction of Orai1 protein. In support of involvement of reactive oxygen species in the HG effects, hydrogen peroxide (H 2 O 2 ) itself significantly decreased abundance of Orai1 protein and increased the level of ubiquitinated Orai1. Taken together, these results suggest that a short-term HG treatment decreased abundance of Orai1 protein in MCs by promoting the protein degradation through the ubiquitination-proteasome and –lysosome mechanisms. This HG-stimulated posttranslational mechanism was mediated by H 2 O 2 .

AB - The short-term effect of high-glucose (HG) treatment on store-operated Ca 2+ entry in mesangial cells (MCs) is not well-known. The aim of the present study was to determine whether and how HG treatment for a short period altered protein abundance of Orai1, the channel mediating store-operated Ca 2+ entry in MCs. Rat and human MCs were exposed to HG (25 mM) for 2, 4, 8, and 24 h, and the abundance of Orai1 protein was significantly decreased at the time points of 8 and 16 h. Consistently, HG treatment for 8 h significantly reduced store-operated Ca 2+ entry in rat MCs. However, HG treatment for the same time periods did not alter the levels of Orai1 transcript. Cycloheximide, a protein synthesis inhibitor, did not affect the HG-induced decrease of Orai1 protein, suggesting a posttranslational mechanism was involved. However, the HG effect on Orai1 protein was significantly attenuated by MG132 (a ubiquitin-proteasome inhibitor) and NH 4 Cl (a lysosomal pathway inhibitor). Furthermore, HG treatment for 8 h stimulated ubiquitination of Orai1 protein. We further found that polyethylene glycol-catalase, an antioxidant, significantly blunted the HG-induced reduction of Orai1 protein. In support of involvement of reactive oxygen species in the HG effects, hydrogen peroxide (H 2 O 2 ) itself significantly decreased abundance of Orai1 protein and increased the level of ubiquitinated Orai1. Taken together, these results suggest that a short-term HG treatment decreased abundance of Orai1 protein in MCs by promoting the protein degradation through the ubiquitination-proteasome and –lysosome mechanisms. This HG-stimulated posttranslational mechanism was mediated by H 2 O 2 .

KW - Hydrogen peroxide

KW - Mesangial cells

KW - Orai1

KW - Store-operated Ca entry

KW - Ubiquitination

UR - http://www.scopus.com/inward/record.url?scp=85059885958&partnerID=8YFLogxK

U2 - 10.1152/ajprenal.00513.2017

DO - 10.1152/ajprenal.00513.2017

M3 - Article

VL - 314

SP - F855-F863

JO - American Journal of Physiology - Renal Physiology

JF - American Journal of Physiology - Renal Physiology

SN - 0363-6127

IS - 5

ER -