Effects of mitochondrial aldehyde dehydrogenase 2 on autophagy-associated proteins in neonatal rat myocardial fibroblasts cultured in high glucose.
10.12122/j.issn.1673-4254.2019.05.04
- Author:
Bi TANG
1
;
Pinfang KANG
1
;
Jianlu GUO
1
;
Lei ZHU
1
;
Qingmei XU
1
;
Qin GAO
2
;
Heng ZHANG
1
;
Hongju WANG
1
Author Information
1. Department of Cardiovascular Medicine, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China.
2. Department of Physiology Cardiovascular Research Center of BengBu Medical College, Bengbu 233030, China.
- Publication Type:Journal Article
- Keywords:
autophagy;
high glucose;
mitochondrial acetaldehyde dehydrogenase 2
- MeSH:
Aldehyde Dehydrogenase;
Aldehyde Dehydrogenase, Mitochondrial;
metabolism;
Animals;
Animals, Newborn;
Autophagy;
Beclin-1;
physiology;
Fibroblasts;
Glucose;
Microtubule-Associated Proteins;
Mitochondrial Proteins;
Rats;
Rats, Sprague-Dawley
- From:
Journal of Southern Medical University
2019;39(5):523-527
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate whether autophagy mediates the effects of aldehyde dehydrogenase 2 (ALDH2) on the proliferation of neonatal rat cardiac fibroblasts cultured in high glucose.
METHODS:Cardiac fibroblasts were isolated from neonatal (within 3 days) SD rats and subcultured. The fibroblasts of the third passage, after identification with immunofluorescence staining for vimentin, were treated with 5.5 mmol/L glucose (control group), 30 mmol/L glucose (high glucose group), or 30 mmol/L glucose in the presence of Alda-1 (an ALDH2 agonist), daidzin (an ALDH2 2 inhibitor), or both. Western blotting was employed to detect ALDH2, microtubule-associated protein 1 light chain 3B subunit (LC3B) and Beclin-1 in the cells, and a hydroxyproline detection kit was used for determining hydroxyproline content in cell culture medium; CCK- 8 kit was used for assessing the proliferation ability of the cardiac fibroblasts after the treatments.
RESULTS:Compared with the control cells, the cells exposed to high glucose exhibited obviously decreased expressions of ALDH2, Beclin-1 and LC3B and increased cell number and hydroxyproline content in the culture medium. Treatment of the high glucose-exposed cells with Alda-1 significantly increased Beclin-1, LC3B, and ALDH2 protein expressions and lowered the cell number and intracellular hydroxyproline content, whereas the application of daidzin resulted in reverse changes in the expressions of ALDH2, Beclin-1 and LC3B, viable cell number and intracellular hydroxyproline content in high glucose-exposed cells.
CONCLUSIONS:Mitochondrial ALDH2 inhibits the proliferation of neonatal rat cardiac fibroblasts induced by high glucose, and the effect is possibly mediated by the up-regulation of autophagy-related proteins Beclin-1 and LC3B.
