ULK1-Regulated Autophagy: A Mechanism in Cellular Protection for ALDH2 against Hyperglycemia.

ULK1-Regulated Autophagy: A Mechanism in Cellular Protection for ALDH2 against Hyperglycemia.

Toxicol Lett. 2017 Nov 08;:

Authors: Liu M, Lu S, He W, Zhang L, Ma Y, Lv P, Ma M, Yu W, Wang J, Zhang M, Zhang Y, Li Y

Abstract
Mitochondrial aldehyde dehydrogenase 2 (ALDH2), an important enzyme in the elimination of toxic aldehydes, is involved in cardioprotection against diabetes mellitus. This study was designed to examine the mechanism behind ALDH2-offered protection against high glucose exposure with a focus on autophagy. H9C2 cells were cultured with normal or high glucose medium in the presence or absence of the ALDH2 agonist Alda-1. GFP-LC3 puncta and immunofluorescence were employed to assess autophagosome formation. Western blotting was applied to evaluate autophagy protein markers Atg5, LC3, p62, ULK1 phosphorylation and ALDH2. JC-1 staining was used to monitor mitochondrial membrane potential and mitochondrial injury. CCK-8 and TUNEL assays were employed for apoptosis and cell viability. Our results indicated that high glucose promoted cell death and decreased cell viability. Levels of autophagy protein marker Atg5, and LC3B were decreased and level of p62 was elevated in hyperglycemic condition, the effects of which were reversed by ALHD2. High glucose lowered mitochondrial membrane potential, the effect of which was accentuated by ULK1 knock-down. All these high glucose-induced responses were negated by Alda-1 along with upregulated autophagy. The autophagy inhibitor 3-MA and lysosomal inhibitor bafilomycin A1 cancelled off whereas autophagy inducer rapamycin mimicked the Alda-1-offered protection against high glucose. High glucose suppressed phosphorylation of ULK1, the effect of which was mitigated by Alda-1. Knock-down of ULK1 using siRNA negated Alda-1-induced upregulation of autophagosome accumulation and LC3 expression. High glucose-dampened autophagy was also confirmed using GFP-LC3 puncta, and immunofluorescence. Taken together, these data suggested that ULK1 played a crucial role in ALDH2-offered protective effect against high glucose exposure-induced cardiomyocyte injury through regulation of autophagy.

PMID: 29128638 [PubMed - as supplied by publisher]

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