Empagliflozin and sacubitril/valsartan reverse methotrexate cardiotoxicity by repressing oxidative stress and hypoxia in heart embryonic H9c2 cardiomyocytes - the role of morphology of mitochondria observed on electron microscopy

by Selleckchem | Jul 12 2023

Objective: Oxidative stress and hypoxia play an important role in the pathogenesis of various cardiovascular diseases. We aimed to evaluate the effectiveness of sacubitril/valsartan (S/V) and Empagliflozin (EMPA) on hypoxia-inducible factor-1α (HIF-1α) and oxidative stress in H9c2 rat embryonic cardiomyocyte cells.

Materials and methods: BH9c2 cardiomyocyte cells were treated with methotrexate (MTX) (10-0.156 μM), empagliflozin (EMPA; 10-0.153 µM) and sacubitril/valsartan (S/V; 100-1.062 µM) for 24, 48 and 72 h. The half maximum inhibitory concentration (IC50) and half maximum excitation concentration (EC50) values of MTX, EMPA and S/V were determined. The cells under investigation were exposed to 2.2 μM MTX before treatment with 2 μM EMPA and 25 μM S/V. The cell viability, lipid peroxidation, oxidation of proteins and antioxidant parameters were measured while morphological changes were also observed by transmission electron microscopy (TEM).

Results: The results showed that treatment with 2 µM EMPA, 25 µM S/V or their combination produced a protective effect against the reduction in cell viability caused by 2.2 µM MTX. While HIF-1α levels plunged to their lowest with S/V treatment, oxidant parameters dipped, and antioxidant parameters soared to their highest level with S/V and EMPA combination treatment. A negative correlation was found between HIF-1α and total antioxidant capacity in the S/V treatment group.

Conclusions: A significant decrease in HIF-1α and oxidant molecules together with an enhancement in antioxidant molecules and normalization of the mitochondria morphology as observed on electron microscopy in S/V and EMPA-treated cells were detected. Although S/V and EMPA have both protective effects against cardiac ischemia and oxidative damage, this effect may be increased more with S/V treatment alone compared to combined treatment.

Comments:

The objective of the study was to evaluate the effectiveness of sacubitril/valsartan (S/V) and empagliflozin (EMPA) in reducing hypoxia-inducible factor-1α (HIF-1α) levels and oxidative stress in H9c2 rat embryonic cardiomyocyte cells.

The researchers conducted the study using H9c2 cardiomyocyte cells and treated them with different concentrations of methotrexate (MTX), empagliflozin, and sacubitril/valsartan for varying durations. They determined the half maximum inhibitory concentration (IC50) and half maximum excitation concentration (EC50) values of MTX, EMPA, and S/V. The cells were exposed to 2.2 μM MTX before treatment with 2 μM EMPA and 25 μM S/V. The researchers measured cell viability, lipid peroxidation, protein oxidation, and antioxidant parameters. They also observed morphological changes using transmission electron microscopy (TEM).

The results showed that treatment with 2 μM EMPA, 25 μM S/V, or their combination protected against the reduction in cell viability caused by 2.2 μM MTX. Treatment with S/V resulted in the lowest levels of HIF-1α, while oxidant parameters decreased and antioxidant parameters increased the most with the combination of S/V and EMPA. In the S/V treatment group, a negative correlation was found between HIF-1α levels and total antioxidant capacity.

The researchers concluded that S/V and EMPA treatment significantly decreased HIF-1α levels and oxidative stress markers while increasing antioxidant parameters. They also observed normalization of mitochondria morphology with S/V and EMPA treatment. Both S/V and EMPA showed protective effects against cardiac ischemia and oxidative damage, but the protective effect may be greater with S/V treatment alone compared to combined treatment.

In summary, the study suggests that S/V and EMPA have a beneficial effect on reducing oxidative stress and HIF-1α levels in cardiomyocyte cells. However, further research is needed to understand the precise mechanisms and potential synergistic effects of S/V and EMPA in cardiovascular diseases.