title: Evaluating the inhibitory potential of Withania somnifera on platelet aggregation and inflammation enzymes: An in vitro and in silico study.
abstract: Context Withania somnifera (L.) Dunal is traditionally used for treating various ailments, but lacks scientific evaluation. Objective This study evaluates Withania somnifera (WS) for its effect on platelet activity and inflammatory enzymes. Materials and methods Aqueous and ethanolic (1:1) leaf extracts were subjected to in vitro indirect haemolytic activity using Naja naja venom, human platelet aggregation was quantified for lipid peroxidation using arachidonic acid (AA) as agonist and 5-lipoxygenase (5-LOX) levels were determined using standard spectrometric assays. Further, molecular docking was performed by the ligand fit method using molegro software package (Molegro ApS, Aarhus, Denmark). Results The study found that aqueous and ethanol extracts have very negligible effect (15%) with an IC50 value of 13.8 mg/mL on PLA2 from Naja naja venom. Further, extracts of WS also had very little effect (18%) with an IC50 value of 16.6 mg/mL on malondialdehyde (MDA) formation. However, a 65% inhibition of 5-LOX with an IC50 value of 0.92 mg/mL was observed in 1:1 ethanol extracts. The same was evident from SAR model with the active ingredient withaferin A binding predominantly on Phe 77, Tyr 98, Arg 99, Asp 164, Leu 168, Ser 382, Arg 395, Tyr 396 and Tyr 614 with an atomic contact energy value of -128.96 compared to standard phenidone (-103.61). Thus, the current study validates the application of WS for inflammatory diseases. Conclusion This study reveals the inhibitory potential of W. somnifera on inflammatory enzymes and platelet aggregation. Thus, WS can serve as a newer, safer and affordable medicine for inflammatory diseases.
authors.affiliation: a Department of Commerce and Science , JSS College of Arts , Mysore , Karnataka , India ;
authors.forename: Madhusudan
authors.initials: M
authors.lastname: M
authors.affiliation: b Department of Studies in Biotechnology, Biochemistry and Microbiology , Mahajana Research Foundation, SBRR Mahajana First Grade College, Affiliated to University of Mysore , Mysore , Karnataka , India ;
authors.forename: Farhan
authors.initials: F
authors.lastname: Zameer
authors.affiliation: c Central Food Technological Research Institute (CFTRI) , Mysore , Karnataka , India ;
authors.forename: Akhilender
authors.initials: A
authors.lastname: Naidu
authors.affiliation: d Department of Biotechnology , Sri Jayachamarajendra College of Engineering, JSS Institution Camp , Mysore , Karnataka , India ;
authors.forename: Nagendra Prasad
authors.initials: NP
authors.lastname: M N
authors.affiliation: e Centre for Emerging Technologies , Jain Global Campus, Jain University , Ramanagara , Karnataka , India ;
authors.forename: Bhadrapura Lakkappa
authors.initials: BL
authors.lastname: Dhananjaya
authors.affiliation: f Department of Studies in Biotechnology, Biochemistry and Microbiology , Pooja Bhagavat Memorial Mahajana Post Graduate Centre, Mahajana Research Foundation, Affiliated to University of Mysore , Mysore , Karnataka , India.
authors.forename: Raghavendra
authors.initials: R
authors.lastname: Hegdekatte
mesh_terms: D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology; D016861:Cyclooxygenase Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D004546:Elapid Venoms / Q000201:enzymology; D000431:Ethanol / Q000737:chemistry; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D016859:Lipoxygenase Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D062105:Molecular Docking Simulation; D015394:Molecular Structure; D064801:Phospholipase A2 Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D054497:Phospholipases A2, Secretory / Q000378:metabolism; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D018515:Plant Leaves; D010946:Plants, Medicinal; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D012997:Solvents / Q000737:chemistry; D013329:Structure-Activity Relationship; D032341:Withania / Q000737:chemistry; D054358:Withanolides / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000893:Anti-Inflammatory Agents; D016861:Cyclooxygenase Inhibitors; D004546:Elapid Venoms; D016859:Lipoxygenase Inhibitors; D064801:Phospholipase A2 Inhibitors; D010936:Plant Extracts; D010975:Platelet Aggregation Inhibitors; D012997:Solvents; D054358:Withanolides; D000431:Ethanol; D054497:Phospholipases A2, Secretory; C009684:withaferin A
keywords: 5-lipoxygenase; Cyclooxygenase; Naja naja venom; Solanaceae; phospholipases; structure–activity relationship; withaferin-A