Study of impact of Wolbachia surface protein on reactive oxygen species production in HepG2 cell line exposed to ethanol

Objective: To study the impact of Wolbachia surface protein (WSP) on reactive oxygen species (ROS) level inethanol (EtOH)-exposed HepG2 cells.Materials and Methods: Increase in ROS level was induced in HepG2 cells by subjecting HepG2 cells toEtOH exposure. Impact of WSP on ROS level was examined by staining of intracellular ROS in cells usingthe specific ROS-detecting dye 2ʹ, 7ʹ-dichlorodihydrofluorescein diacetate (H2DCFDA), followed by flowcytometric analysis.Results and Conclusion: Flow cytometry analysis using H2DCFDA-based staining study of ROS level inHepG2 cells revealed that EtOH caused oxidative stress in HepG2 cells by inducing production of high levelsof ROS. However, EtOH-induced increased ROS production in cells decreased with treatment of WSP.From the current study, we can culminate that WSP provides cytoprotective action against EtOH-inducedincreased ROS production and oxidative stress in HepG2 cells by decreasing ROS production. This will beof significance for the treatment of EtOH-related liver ailments. Thus, this article emphasizes that WSP withprotecting ability could be used as a powerful therapeutic drug to treat EtOH-related liver ailments.

Molecular Dynamics, Simulation of Protein-Protein Complex and their Role in Cytoprotective Process in Ethanol-induced Toxicity of HepG2 Cell Line

Aim: To examine the protein-protein interaction of Wolbachia Surface Protein (WSP of Uzifly) with six proteins involved in Ethanol-induced toxicity and the proteins involved in its cytoprotective process in HepG2 cell line (CYP2E1, Superoxide dismutase, Catalase, Death-associated protein kinase1, Alcohol dehydrogenases (Alpha/beta/gamma) and Cytochrome-C) and to study real time molecular dynamics. Methodology: Modelled structure of WSP of Uzifly was retrieved from our laboratory archive. The proteins involved in the Ethanol-induced toxicity and the proteins involved in its cytoprotective process in HepG2 cell line were chosen based on the literature study. The six proteins like CYP2E1, Superoxide dismutase, Catalase, Death-associated protein kinase1, Alcohol dehydrogenases (Alpha/beta/gamma) and Cytochrome-C which are involved in the Ethanol-induced toxicity and the proteins involved in its cytoprotective process in HepG2 cell line were retrieved from PDB database with ID: PDB (3T3Z), PDB (2C9V), PDB (1DGG), PDB (2YAK), PDB (1U3W) and PDB (3NWV) respectively. Docking study was processed using ZDOCK and the best poses of protein were sorted using rDock. Finally, the atomic level interaction was studied for the best-scored protein-protein complex. The best complex was further subjected to molecular dynamics simulation to study its stability using standard dynamics cascade tool. Results: From the results, it was observed that three proteins such as Cytochrome-C, CYP2E1 and Superoxide dismutase have more favourable shape complementarity for WSP binding to exhibit the cytoprotective process. However, the interaction analysis was done only for the top complex, Cytochrome-C-WSP. Time dependent parameter analysis of best complex Cytochrome-C-WSP showed that root-mean-square deviation (RMSD) values initially deviated but it was stabilized at the end of 1ns dynamics. The radius of gyration (Rg) during dynamics was within the limit. Conclusion: This insilico study revealed that WSP has cytoprotective potential and therapeutical application.