Objectives To examine the effect of ginsenoside Re (G-Re) on autophagy in H9c2 cardiomyocyte cells under starvation.
Methods H9c2 cells were cultured in glucose-free and serum-free medium for several hours as the glucose deprivation (GD) stimulation. To measure the amount of autophagosome formation, we determined the membrane-bound form of autophagy-related protein LC3B-2 by immunoblotting. To evaluate the situation of autophagy flux, we added 10 nmol/l bafilomycin A1 (BafA1) into the medium to block the fusion processes between autophagosomes and lysosomes. H9c2 cells under GD were treated with 100 µmol/l G-Re. LC3B-2 measurement and immunofluorescence were conducted to display the effect of G-Re on autophagy in these cells. Cell viability, ATP content, malondialdehyde level and superoxide dismutase activity in cultured medium were determined to appreciate the physiological relevance of autophagy changes due to G-Re treatment. We also assayed phosphorylated AMPKα and mTOR to explore the mechanisms underlying the effect of G-Re on autophagy in cells under GD.
Results In H9c2 cells under GD, LC3B-2 increased in a time-dependent manner in association with the decrease of cell viability and cellular ATP content. In H9c2 cell under GD and treated with 100 µmol/l G-Re, LC3B-2 expression decreased, accompanied by the rescue of cell death, the increase of cellular ATP content, and the relief of oxidative stress. The higher p-AMPKα in H9c2 cell under GD decreased when they were treated with 100 µmol/l G-Re, probably relating to the mechanisms underlying the inhibition of autophagosomal formation by G-Re.
Conclusions Starvation induced autophagy in H9c2 cells and led to cell injury. Treatment of 100 µmol/l G-Re inhibited autophagosomal formation in the cells, which may be beneficial to the cardiomyocytes under starvation.
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.