In silico and In vivo Studies of Calotropis procera leaf and root extracts on Mitochondrial-Related parameters in Wistar Rats

Drugs targeting Mitochondrial Membrane Permeability Transition (MMPT) pore opening are of great interest for conditions arising from apoptosis dysregulation. This study investigate MMPT pore inducing effect of Calotropis procera leaves and root extracts on mitochondrial-related parameters in the liver of healthy male Wistar rats. Extraction was done by standard methods, using Ethylacetate and Butanol solvent. Fifty–two rats weighing between 130 g - 150 g were divided into thirteen groups (n=4). Group 1 received distilled water, other groups were administered ethylacetate leaf extract (ELE), butanol leaf extract (BLE), ethylacetate root extract (ERE) and butanol root extracts (BRE). Liver Mitochondrial pore opening, mitochondrial ATPase activity and lipid peroxidation were assessed spectrophotometrically, while the interaction between human Cyclophilin D (a pore activating protein with PDB ID: 2BIT) and identified phytochemicals of Calotropis procera leaves and roots extracts were studied In-silico. Varying concentrations of the extracts induced MMPT pore opening by 6.6, 7.4 and 5.9 folds for BLE and 10.2, 7.3 and 6.4 folds for ELE at 40, 50 and 60 mg/100g bw respectively when compared with control group. Root extracts showed significant MMPT pore opening with a fold increase of 12.2, 10.1 and 20.4 for BRE and 11.37, 11.84, for ERE at at 40 , 50 and 60 mg/100g bw respectively. BLE, BRE, ELE and ERE showed increase in ATPase activities with respect to the control group. Malondialdehyde (MDA) levels as an indication of lipid peroxidation increased significantly when compared with control. Molecular docking and simulation showed the existence of stable interactions between human Cyclophilin D with phytochemicals of Calotropis procera with 2‶-Oxovoruscharin having highest binding affinity of -7.9 kcal/mol. C. procera has potential for therapeutic use for the treatment of disorders related to derangement of