PUBLICATION
Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes
- Authors
- Meng, X.H., Chen, B., Zhang, J.P.
- ID
- ZDB-PUB-170920-8
- Date
- 2017
- Source
- International journal of biological sciences 13: 985-995 (Journal)
- Registered Authors
- Zhang, Jing-pu
- Keywords
- animal models, defective autophagy, insulin-targeting tissues, preproinsulin, subcellular localization, type 2 diabetes
- MeSH Terms
-
- Animals
- Autophagy/physiology*
- Autophagy-Related Proteins/metabolism
- Blotting, Western
- Diabetes Mellitus, Type 2/metabolism*
- Glucose Transporter Type 2/metabolism
- Hep G2 Cells
- Humans
- Immunoprecipitation
- Insulin/metabolism*
- Protein Precursors/metabolism
- Real-Time Polymerase Chain Reaction
- Zebrafish
- PubMed
- 28924380 Full text @ Int. J. Biol. Sci.
Citation
Meng, X.H., Chen, B., Zhang, J.P. (2017) Intracellular Insulin and Impaired Autophagy in a Zebrafish model and a Cell Model of Type 2 diabetes. International journal of biological sciences. 13:985-995.
Abstract
Type 2 diabetes mellitus is characterized by insulin resistance. However, the complete molecular mechanism remains unclear. In this study, zebrafish were fed a long-term high-fat diet to induce type 2 diabetes, which resulted in a higher body weight, body mass index, more lipid vacuoles in liver, increased insulin transcription level in liver, brain and muscle, and high fasting blood glucose in the high-fat diet zebrafish. Oppositely, the transcription levels of insulin substrate-2 and glucose transporter 2 were significantly decreased, indicating insulin signaling pathway and glucose transport impaired in the insulin-targeting tissues. Transcription of the autophagy-related genes, ATG3, ATG4B, ATG5, ATG7, ATG12, and FOXO3, were decreased but autophagy inhibitor gene m-TOR increased, and autophagy-flux was inhibited in liver of the high-fat diet zebrafish. Main of these changes were confirmed in palmitic acid-treated HepG2 cells. Further, in co-immunoprecipitation and subcellular co-localization experiments, the conjunction of preproinsulin with cargo-recognition protein p62 increased, but conjuncts of autophagosome with p62-cargo, lysosomes with p62-cargo, and autolysosomes decreased apparently. Interestingly, lysosomes, autolysosomes and conjuncts of p62-insulin localized at the periphery of palmitic acid-treated cells, the margination of lysosomes may mediate deactivation of proteases activity. These findings suggest that intracellular high-lipid may trigger defective autophagy, defective downstream signaling of insulin and accumulated intracellular preproinsulin, leading to dysregulation of cell homeostasis mechanism, which may be one of reasons involved in insulin-resistance in type 2 diabetes.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping