SEAP activity serves for demonstrating ER stress induction by glucolipotoxicity as well as testing ER stress inhibitory potential of therapeutic agents

Lenin, R and Mohan, V and Balasubramanyam, M (2015) SEAP activity serves for demonstrating ER stress induction by glucolipotoxicity as well as testing ER stress inhibitory potential of therapeutic agents. Molecular and Cellular Biochemistry, 404 (1-2). p. 271. ISSN 0300-8177

[img]PDF
Restricted to MDRF users only. Others may ->

745Kb

Abstract

Endoplasmic reticulum (ER) stress is emerging as a unifying paradigm and one of the underlying mechanisms in the genesis of diabetes and its complications. While this has prompted the development of ER stress inhibitors, there is a limitation in monitoring of ER stress in vitro and in vivo by reliable methodologies. We validated the secreted alkaline phosphatase (SEAP) activity as a surrogate marker of ER stress in mouse β-TC6 cells exposed to glucolipotoxicity or tunicamycin and studied insulin secretion along with alterations in ER stress markers. SEAP activity assay was measured using the Great EscAPe SEAP kit, insulin levels were determined by Mercodia reagents and mRNA expression of ER stress markers was quantified by real-time PCR. SEAP activity in β-cells was significantly decreased (indicating increased ER stress) on exposure either to glucolipotoxicity or tunicamycin. This was accompanied by an increased mRNA expression of ER stress markers (GRP-78, PERK, IRE1α, ATF6, XBP-1, and CHOP) and decreased insulin secretion. Treating the cells with phenylbutyric acid normalized SEAP activity, decreased mRNA expression of ER stress markers and improved insulin secretion. Interestingly, cells exposed to different classes of anti-diabetes agents or compounds such as resveratrol resisted ER stress. Methylglyoxal also induces ER stress and this was counteracted by aminoguanidine. Out study demonstrates SEAP activity as a novel ER stress monitoring assay to investigate the therapeutic value of agents with ER stress inhibitory potential. Future studies should focus on the exercise of adopting this reporter assay for high-throughput screening mode of drug discovery.

Item Type:Article
Official URL/DOI:http://dx.doi.org/10.1007/s11010-015-2387-1
Uncontrolled Keywords:ER stress; SEAP activity; UPR;Glucolipotoxicity; PBA; Metformin; Vildagliptin; b-Cells; Abbre
Subjects:Biochemistry,Cell and Molecular Signalling
Diabetes
Divisions:Department of Cell and Molecular Biology
Department of Diabetology
Department of Advanced Research Biochemistry
ID Code:906
Deposited By:surendar radha
Deposited On:04 May 2015 17:25
Last Modified:04 May 2015 17:25

Repository Staff Only: item control page