In Silico Studies and in Vitro Modulatory Effects of Kolaviron on Rat Liver Mitochondrial Membrane Permeability Transition Pore

Modulatory effects of varying concentrations of kolaviron was investigated on isolated rat liver mitochondrial membrane permeability transition (mPT) pore and in silico apoptotic markers. mPT pore was estimated by the extent of mitochondrial swelling using spectroscopy. Molecular docking was carried out via PyRx software tool. The results showed that there was no significant change in absorbance after energizing the mitochondria in the absence of Ca 2+ . There was large amplitude opening of the intact mitochondrial pore when challenged with Calcium. However, there was reversal of pore-opening by standard inhibitor, spermine (63% inhibition). In vitro results revealed that varying concentrations of kolaviron did not exert effect on mPT pore opening in the absence of Ca 2+ . But in the presence of Ca 2+ , kolaviron reversed Ca 2+ -induced mPT pore opening in concentration-dependent manner (0.05µg/ml < 0.5µg/ml < 1.5µg/ml < 3µg/ml < 6µg/ml). There was also a concentration-dependent Fe 2+ -induced lipid peroxidation inhibition by kolaviron. In addition, kolaviron did not trigger ATPase activity. In in silico studies, it was observed that there were strong interactions of ANT, VDAC, cyclophilin D, MDM2, p53, cytochrome c and Bax with kolaflavanone, GB1 and GB2 (kolaviron components) while BCL-2 exhibited weak binding energy with the bioactive agents relative to the respective standard inhibitors. The above findings suggest that kolaviron bioactive compounds may be excellent agents in modulating mitochondrial membrane permeability transition pore through interaction with several pore components and apoptogenic proteins.