Jianlu Guo1, Pinfang Kang1, Lei Zhu1, Shuo Sun1, Min Tao2, Heng Zhang1, Bi Tang1. 1. Department of Cardiology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China. 2. Department of Cardiology, Huishan District People's Hospital, Wuxi 214100, China.
Abstract
OBJECTIVE: To investigate whether CaN-NFAT3 pathway mediates the protective effects of aldehyde dehydrogenase (ALDH) 2 in high glucose-treated neonatal rat ventricular myocytes. METHODS: The ventricular myocytes were isolated from the heart of neonatal (within 3 days) SD rats by enzyme digestion and cultured in the presence of 5-Brdu. After reaching confluence, the cultured ventricular myocytes were identified using immunofluorescence assay for α-SA protein. The cells were then cultured in either normal (5 mmol/L) or high glucose (30 mmol/L) medium in the presence of ALDH2 agonist Alda-1, ALDH 2 inhibitor Daidzin, or Alda-1 and NFAT3 inhibitor (11R-VIVIT). Fluorescent probe and ELISA were used to detect intracellular Ca2+ concentration and CaN content, respectively; ALDH2, CaN and NFAT3 protein expressions in the cells were detected using Western blotting. RESULTS: Compared with cells cultured in normal glucose, the cells exposed to high glucose showed a significantly decreased expression of ALDH2 protein (P < 0.05) and increased expressions of CaN (P < 0.05) and NFAT3 proteins with also increased intracellular CaN and Ca2+ concentrations (P < 0.01). Alda-1 treatment significantly lowered Ca2+ concentration (P < 0.05), intracellular CaN content (P < 0.01), and CaN and NFAT3 protein expressions (P < 0.05), and increased ALDH2 protein expression (P < 0.05) in high glucose- exposed cells; Daidzin treatment significantly increased Ca2+ concentration (P < 0.01) and intracellular CaN content (P < 0.05) in the exposed cells. Compared with Alda-1 alone, treatment of the high glucose-exposed cells with both Alda-1 and 11R-VIVIT did not produce significant changes in the expression of ALDH2 protein (P>0.05) but significantly reduced the expression of NFAT3 protein (P < 0.05). CONCLUSIONS: Mitochondrial ALDH2 protects neonatal rat cardiomyocytes against high glucose-induced injury possibly by negatively regulating Ca2+-CaN-NFAT3 signaling pathway.
OBJECTIVE: To investigate whether CaN-NFAT3 pathway mediates the protective effects of aldehyde dehydrogenase (ALDH) 2 in high glucose-treated neonatal rat ventricular myocytes. METHODS: The ventricular myocytes were isolated from the heart of neonatal (within 3 days) SD rats by enzyme digestion and cultured in the presence of 5-Brdu. After reaching confluence, the cultured ventricular myocytes were identified using immunofluorescence assay for α-SA protein. The cells were then cultured in either normal (5 mmol/L) or high glucose (30 mmol/L) medium in the presence of ALDH2 agonist Alda-1, ALDH 2 inhibitor Daidzin, or Alda-1 and NFAT3 inhibitor (11R-VIVIT). Fluorescent probe and ELISA were used to detect intracellular Ca2+ concentration and CaN content, respectively; ALDH2, CaN and NFAT3 protein expressions in the cells were detected using Western blotting. RESULTS: Compared with cells cultured in normal glucose, the cells exposed to high glucose showed a significantly decreased expression of ALDH2 protein (P &lt; 0.05) and increased expressions of CaN (P &lt; 0.05) and NFAT3 proteins with also increased intracellular CaN and Ca2+ concentrations (P &lt; 0.01). Alda-1 treatment significantly lowered Ca2+ concentration (P &lt; 0.05), intracellular CaN content (P &lt; 0.01), and CaN and NFAT3 protein expressions (P &lt; 0.05), and increased ALDH2 protein expression (P &lt; 0.05) in high glucose- exposed cells; Daidzin treatment significantly increased Ca2+ concentration (P &lt; 0.01) and intracellular CaN content (P &lt; 0.05) in the exposed cells. Compared with Alda-1 alone, treatment of the high glucose-exposed cells with both Alda-1 and 11R-VIVIT did not produce significant changes in the expression of ALDH2 protein (P&gt;0.05) but significantly reduced the expression of NFAT3 protein (P &lt; 0.05). CONCLUSIONS: Mitochondrial ALDH2 protects neonatal rat cardiomyocytes against high glucose-induced injury possibly by negatively regulating Ca2+-CaN-NFAT3 signaling pathway.
Entities:
Keywords:
activated T cell nuclear factor 3; calcineurin; high glucose; mitochondrial acetaldehyde dehydrogenase 2
Authors: Sujith Dassanayaka; Yuting Zheng; Andrew A Gibb; Timothy D Cummins; Lindsey A McNally; Kenneth R Brittian; Ganapathy Jagatheesan; Timothy N Audam; Bethany W Long; Robert E Brainard; Steven P Jones; Bradford G Hill Journal: Redox Biol Date: 2018-06-01 Impact factor: 11.799