Author(s)

Keisuke Sato¹, Shoya Endo¹, Natsuki Ota¹, Akira Takaguri², Kumi Satoh², Ryosuke Tatsunami¹

¹Department of Pharmacy, Hokkaido University of Science, Hokkaido, Japan.
²Department of Pharmacology, Hokkaido University of Science, Hokkaido, Japan.

The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay is used as a major method to evaluate cell viability. However, in some cases, the results may reflect mitochondrial status regardless of viability. Epalrestat (EPS) is currently available for the treatment of diabetic neuropathy. In this study, we report that EPS at near-plasma concentrations increases MTS reduction activity independent of cell number in bovine aortic endothelial cells. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a key transcription factor that plays a pivotal role in inducing the expression of genes encoding detoxifying and defensive proteins. Sulforaphane (an Nrf2 activator) also increased MTS-reducing activity, similar to EPS. Knockdown of Nrf2 by short interfering RNA suppressed EPS-induced MTS reduction. These results suggest that EPS increases MTS reduction activity via the Nrf2 pathway. Furthermore, the results that EPS increases ATP production and that electron transfer chain inhibitors suppress EPS-induced MTS reduction activity suggest that EPS may activate mitochondrial status. Because mitochondrial disorders cause numerous diseases, we suggest that EPS has new beneficial properties that may prevent the development and progression of disorders caused by mitochondrial dysfunction.

EPS, MTS, Nrf2, Mitochondria, BAECs

Sato, K. , Endo, S. , Ota, N. , Takaguri, A. , Satoh, K. and Tatsunami, R. (2023) Epalrestat Enhances MTS Reduction Activity Independent of Cell Numbers in Bovine Aortic Endothelial Cells. Pharmacology & Pharmacy, 14, 59-71. doi: 10.4236/pp.2023.143005.