Anti-apoptotic effects of autophagy via ROS regulation in microtubule-targeted and PDGF-stimulated vascular smooth muscle cells.
10.4196/kjpp.2018.22.3.349
- Author:
Hyun Soo PARK
1
;
Joo Hui HAN
;
Sang Hyuk JUNG
;
Do Hyung LEE
;
Kyung Sun HEO
;
Chang Seon MYUNG
Author Information
1. Department of Pharmacology, Chungnam National University College of Pharmacy, Daejeon 34134, Korea. cm8r@cnu.ac.kr
- Publication Type:Original Article
- Keywords:
Apoptosis;
Autophagy;
Proliferation;
Reactive oxygen species;
Vascular smooth muscle cell
- MeSH:
Apoptosis;
Atherosclerosis;
Autophagy*;
Blotting, Western;
Cardiovascular Diseases;
Caspase 3;
Cell Count;
Cell Survival;
Fluorescence;
Fluorescent Antibody Technique;
In Vitro Techniques;
Microtubules;
Muscle, Smooth, Vascular*;
Reactive Oxygen Species;
Stents
- From:The Korean Journal of Physiology and Pharmacology
2018;22(3):349-360
- CountryRepublic of Korea
- Language:English
-
Abstract:
Autophagy has been studied as a therapeutic strategy for cardiovascular diseases. However, insufficient studies have been reported concerning the influence of vascular smooth muscle cells (VSMCs) through autophagy regulation. The aim of the present study was to determine the effects of VSMCs on the regulation of autophagy under in vitro conditions similar to vascular status of the equipped microtubule target agent-eluting stent and increased release of platelet-derived growth factor-BB (PDGF-BB). Cell viability and proliferation were measured using MTT and cell counting assays. Immunofluorescence using an anti-α-tubulin antibody was performed to determine microtubule dynamic formation. Cell apoptosis was measured by cleavage of caspase-3 using western blot analysis, and by nuclear fragmentation using a fluorescence assay. Autophagy activity was assessed by microtubule-associated protein light chain 3-II (LC-II) using western blot analysis. Levels of intracellular reactive oxygen species (ROS) were measured using H₂DCFDA. The proliferation and viability of VSMCs were inhibited by microtubule regulation. Additionally, microtubule-regulated and PDGF-BB-stimulated VSMCs increased the cleavage of caspase-3 more than only the microtubule-regulated condition, similar to that of LC3-II, implying autophagy. Inhibitory autophagy of microtubule-regulated and PDGF-BB-stimulated VSMCs resulted in low viability. However, enhancement of autophagy maintained survival through the reduction of ROS. These results suggest that the apoptosis of conditioned VSMCs is decreased by the blocking generation of ROS via the promotion of autophagy, and proliferation is also inhibited. Thus, promoting autophagy as a therapeutic target for vascular restenosis and atherosclerosis may be a good strategy.
