title: Therapeutic potential of Taraxacum officinale against HCV NS5B polymerase: In-vitro and In silico study.
abstract: Discovery of alternative and complementary regimens for HCV infection treatment is a need of time from clinical as well as economical point of views. Low cost of bioactive natural compounds production, high biochemical diversity and inexistent/milder side effects contribute to new therapies. Aim of this study is to clarify anti-HCV role of Taraxacum officinale, a natural habitat plant rich of flavonoids. In this study, methanol extract of T. officinale leaves was initially analyzed for its cytotoxic activity in human hepatoma (Huh-7) and CHO cell lines. Hepatoma cells were transfected with pCR3.1/Flagtag/HCV NS5B gene cloned vector (genotype 1a) along with T. officinale extract. Considering NS5B polymerase as potential therapeutic drug target, twelve phytochemicals of T. officinale were selected as ligands for molecular interaction with NS5B protein using Molecular Operating Environment (MOE) software. Sofosbuvir (Sovaldi: brand name) currently approved as new anti-HCV drug, was used as standard in current study for comparative analysis in computational docking screening. HCV NS5B polymerase as name indicates plays key role in viral genome replication. On the basis of which NS5B gene is targeted for determining antiviral role of T. officinale extract and 65% inhibition of NS5B expression was documented at nontoxic dose concentration (200μg/ml) using Real-time PCR. In addition, 57% inhibition of HCV replication was recorded when incubating Huh-7 cells with high titer serum of HCV infected patients along with leaves extract. Phytochemicals for instance d-glucopyranoside (-31.212 Kcal/mol), Quercetin (-29.222 Kcal/mol), Luteolin (-26.941 Kcal/mol) and some others displayed least binding energies as compared to standard drug Sofosbuvir (-21.0746 Kcal/mol). Results of our study strongly revealed that T. officinale leaves extract potentially blocked the viral replication and NS5B gene expression without posing any toxic effect on normal fibroblast cells of body.
authors: Department of Environmental Sciences, COMSATS Institute of Information Technology, Abbottabad, Pakistan. Electronic address: sidrarehman@ciit.net.pk.
authors: Sidra
authors: S
authors: Rehman
authors: Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
authors: Bushra
authors: B
authors: Ijaz
authors: Department of Pharmacy, COMSATS Institute of Information Technology, Abbottabad, Pakistan.
authors: Nighat
authors: N
authors: Fatima
authors: Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, Pakistan.
authors: Syed Aun
authors: SA
authors: Muhammad
authors: Dean Post Graduate Study, Allama Iqbal Medical College, Lahore, Pakistan.
authors: Sheikh
authors: S
authors: Riazuddin
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D001665:Binding Sites; D016466:CHO Cells; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003198:Computer Simulation; D006224:Cricetinae; D003412:Cricetulus; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D016174:Hepacivirus / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D008024:Ligands; D062105:Molecular Docking Simulation; D064209:Phytochemicals / Q000032:analysis; D010936:Plant Extracts / Q000494:pharmacology*; D031205:Taraxacum / Q000737:chemistry*; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D008024:Ligands; D064209:Phytochemicals; D010936:Plant Extracts; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
keywords: Anti-HCV; Hepatitis C virus; In-vitro; Molecular docking; NS5B polymerase; Taraxacum officinale