Inhibition of Galectin-3 Alleviates Cigarette Smoke Extract-Induced Autophagy and Dysfunction in Endothelial Progenitor Cells

Autor: Lu Fang, Xiaoyan Wang, Shu Meng, ChongZhe Pei, Lin Yanjun
Rok vydání: 2019
Předmět:
0301 basic medicine
Autophagosome
Aging
Article Subject
Angiogenesis
Galectin 3
Neovascularization
Physiologic

Biochemistry
Cigarette Smoking
03 medical and health sciences
0302 clinical medicine
AMP-Activated Protein Kinase Kinases
Downregulation and upregulation
Cell Movement
Enos
parasitic diseases
Autophagy
Humans
Viability assay
RNA
Small Interfering

lcsh:QH573-671
Progenitor cell
Cells
Cultured

Endothelial Progenitor Cells
Tube formation
biology
lcsh:Cytology
Chemistry
TOR Serine-Threonine Kinases
RNA-Binding Proteins
Cell Differentiation
Tobacco Products
Cell Biology
General Medicine
biology.organism_classification
Cell biology
Oxidative Stress
030104 developmental biology
Gene Expression Regulation
030220 oncology & carcinogenesis
embryonic structures
cardiovascular system
Protein Kinases
Research Article
Signal Transduction
circulatory and respiratory physiology
Zdroj: Oxidative Medicine and Cellular Longevity, Vol 2019 (2019)
Oxidative Medicine and Cellular Longevity
ISSN: 1942-0994
1942-0900
DOI: 10.1155/2019/7252943
Popis: Endothelial progenitor cells (EPCs) have the potential to repair damaged blood vessels and promote angiogenesis. Smoking, an important risk factor for cardiovascular diseases, is associated with impaired functions of EPCs. However, the underlying mechanisms remain unclear. The aim of the study was to investigate the effects of cigarette smoke extract (CSE) on autophagy and dysfunction of EPCs and the involvement of galectin-3 in its effects. EPCs were treated with 8% CSE for 24 h (without affecting cell viability). EPC functions were assessed by tube formation and migration capacity and intracellular ROS and eNOS expression. Autophagy was assessed by autophagic protein expression by Western blotting and immunofluorescence microscopy and autophagosome accumulation by transmission electron microscopy. Galectin-3 expression was measured by real-time PCR, Western blotting, and immunofluorescence microscopy, while phospho-AMPK and phospho-mTOR were measured by Western blotting. EPCs were transfected by shRNA-Gal-3 or shRNA-NC before treatment with CSE to examine the effects of galectin-3 on CSE-induced autophagy and dysfunction of EPCs. CSE-treated EPCs showed decreased tube formation and migration ability and eNOS expression but increased oxidative stress. CSE also induced autophagy which was characterized by a decrease in p62 protein, an increase in LC3B-II/I ratio, and accumulation of autophagosomes. CSE upregulated galectin-3 expression on EPCs. Inhibition of galectin-3 abrogated CSE-induced autophagy and dysfunction of EPCs. CSE activated phospho-AMPK and inhibited phospho-mTOR, and inhibition of galectin-3 abolished CSE’s effect on activating phospho-AMPK and inhibiting phospho-mTOR. In conclusion, our results suggest that galectin-3 mediates CSE-induced EPC autophagy and dysfunction, likely via the AMPK/mTOR signaling pathway.
Databáze: OpenAIRE