Mitochondrial dysfunction in H9c2 cells during ischemia and amelioration withTribulus terrestrisL.

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Abstract

Aims:

The present study investigates the protective effect of partially characterized Tribulus terrestris L. fruit methanol extract against mitochondrial dysfunction in cell based (H9c2) myocardial ischemia model.

Main methods:

To induce ischemia, the cells were maintained in an ischemic buffer (composition in mM − 137 NaCl, 12 KCl, 0.5 MgCl2, 0.9 CaCl2, 20 HEPES, 20 2-deoxy-d-glucose, pH − 6.2) at 37 °C with 0.1% O2, 5% CO2, and 95% N2 in a hypoxia incubator for 1 h. Cells were pretreated with various concentrations of T. terrestris L. fruit methanol extract (10 and 25 μg/ml) and Cyclosporin A (1 μM) for 24 h prior to the induction of ischemia.

Key findings:

Different parameters like lactate dehydrogenase release, total antioxidant capacity, glutathione content and antioxidant enzymes were investigated. Studies were conducted on mitochondria by analyzing alterations in mitochondrial membrane potential, integrity, and dynamics (fission and fusion proteins — Mfn1, Mfn2, OPA1, Drp1 and Fis1). Various biochemical processes in mitochondria like activity of electron transport chain (ETC) complexes, oxygen consumption and ATP production was measured. Ischemia for 1 h caused a significant (p ≤ 0.05) increase in LDH leakage, decrease in antioxidant activity and caused mitochondrial dysfunction. T. terrestris L. fruit methanol extract pretreatment was found effective in safeguarding mitochondria via its antioxidant potential, mediated through various bioactives. HPLC of T. terrestris L. fruit methanol extract revealed the presence of ferulic acid, phloridzin and diosgenin.

Significance:

T. terrestris L. fruit ameliorate ischemic insult in H9c2 cells by safeguarding mitochondrial function. This validates the use of T. terrestris L. against heart disorders.

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