In vitro and in silico investigation of garlic’s (Allium sativum) bioactivity against 15-lipoxygenase mediated inflammopathies

Introduction: Garlic (Allium sativum) is widely used as a flavor-enhancing dietary ingredient and exhibits a wide spectrum of pharmacological effects. This study aimed to investigate the therapeutic effects of aqueous garlic extract to explore the bioactivity against 15-lipoxygenase (15-LOX) mediated inflammopathies. Methods: In this study, the antioxidant (DPPH free radical scavenging assay and reducing power assay), anti-inflammatory (hypotonicity-induced hemolysis assay and 15-LOX inhibition assay) and anticoagulation (serine protease inhibition assay and prothrombin time assay) effects of the aqueous garlic extract were investigated. Furthermore, in silico molecular docking and dynamic simulation analysis of reported small compounds of garlic against 15-LOX1 and 15-LOX2 were performed to figure out the most efficient phytochemical ligands and validate the anti-inflammatory potential. Results: The DPPH scavenging effect and the reducing power of the extract were found with the IC50 of 213.87 ± 1.49 μgmL-1 and EC50 of 124.78 ± 3.39 μgmL-1, respectively. In the hypotonicity-induced hemolysis and 15-LOX inhibition assay, the IC50 values were observed as 147.59 ± 2.98 μgmL-1 and 250.05 ± 8.48 μgmL-1, respectively. The extract inhibited serine protease activity with an IC50 of 301.33 ± 1.31 μgmL-1 and prevented blood coagulation for 10.05 ± 0.35 minutes in prothrombin time assay. The in silico study identified Rhamnetin as a potential 15-LOX1 and 15-LOX2 inhibitor, and it exhibited a stable interaction with the targets throughout the 100 ns dynamic simulation. Conclusion: The findings of this study provide molecular insights into garlic’s medicinal properties as well as its bioactive compounds, which can be potential therapeutic interventions for 15-LOX mediated inflammations.

[1]  K. Nagaz,et al.  Biological activities and phenolic compounds of olive oil mill wastewater from Abani, endemic Algerian variety , 2022, Scientific Reports.

[2]  M. Lopes-Pires,et al.  Clotting Dysfunction in Sepsis: A Role for ROS and Potential for Therapeutic Intervention , 2021, Antioxidants.

[3]  S. Lamponi.  Bioactive Natural Compounds with Antiplatelet and Anticoagulant Activity and Their Potential Role in the Treatment of Thrombotic Disorders , 2021, Life.

[4]  Md. Rezanur Rahman,et al.  Analysis of SYK Gene as a Prognostic Biomarker and Suggested Potential Bioactive Phytochemicals as an Alternative Therapeutic Option for Colorectal Cancer: An In-Silico Pharmaco-Informatics Investigation , 2021, Journal of personalized medicine.

[5]  Loleny Tavares,et al.  Bioactive compounds of garlic: A comprehensive review of encapsulation technologies, characterization of the encapsulated garlic compounds and their industrial applicability , 2021 .

[6]  Md. Hafijur Rahman,et al.  Molecular optimization, docking, and dynamic simulation profiling of selective aromatic phytochemical ligands in blocking the SARS-CoV-2 S protein attachment to ACE2 receptor: an in silico approach of targeted drug designing , 2021, Journal of advanced veterinary and animal research.

[7]  C. Yuan,et al.  A general strategy to inhibit serine protease by targeting its autolysis loop , 2021, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  Jinbo Xu,et al.  Deep template-based protein structure prediction , 2020, bioRxiv.

[9]  Asif Ahmed,et al.  Predicting multi-enzyme inhibition in the arachidonic acid metabolic network by Heritiera fomes extracts , 2020, Journal of biomolecular structure & dynamics.

[10]  T. Emran,et al.  Membrane stabilization as a mechanism of the anti-inflammatory activity of ethanolic root extract of Choi (Piper chaba) , 2020, Clinical Phytoscience.

[11]  A. Elengoe,et al.  Molecular Docking of Curcumin With Breast Cancer Cell Line Proteins , 2020, Pharmaceutical and Biomedical Research.

[12]  Md. Emdadul Islam,et al.  Concordance of antioxidant and anti-Inflammatory activity in Xylocarpus granatum (Koen) , 2019 .

[13]  Md. Emdadul Islam,et al.  Antioxidant, Anti-inflammatory, and Anticoagulation Properties of Aegiceras corniculatum and Acanthus ilicifolius , 2019 .

[14]  Md. Emdadul Islam,et al.  Antioxidant and antibacterial activity of three herbs belonging to Zingiber genus of Bangladesh , 2019, Advances in Traditional Medicine.

[15]  Z. Peng,et al.  Reactive Oxygen Species-Induced Lipid Peroxidation in Apoptosis, Autophagy, and Ferroptosis , 2019, Oxidative medicine and cellular longevity.

[16]  D. Patil,et al.  Membrane Stabilization assay for Anti-inflammatory activity yields misleading results for samples containing traces of Methanol , 2019, Asian Journal of Pharmaceutical Research.

[17]  Y. Sultanbawa,et al.  Phytochemical Characteristics and Antimicrobial Activity of Australian Grown Garlic (Allium Sativum L.) Cultivars , 2019, Foods.

[18]  H. Corke,et al.  Bioactive Compounds and Biological Functions of Garlic (Allium sativum L.) , 2019, Foods.

[19]  R. S. Chauhan,et al.  Qualitative phytochemical analysis of Allium sativum (Garlic) and Curcuma longa (Turmeric) , 2019 .

[20]  C. Matar,et al.  The Immunomodulatory and Anti-Inflammatory Role of Polyphenols , 2018, Nutrients.

[21]  Ji-Han Kim,et al.  Antioxidant and antimicrobial activities of fresh garlic and aged garlic by-products extracted with different solvents , 2018, Food Science and Biotechnology.

[22]  S. Hastuti,et al.  The Administration of Garlic Extract on Eimeria stiedai Oocysts and the Hematological Profile of the Coccidia Infected Rabbits , 2017 .

[23]  V. Biju,et al.  In vivo ROS production and use of oxidative stress-derived biomarkers to detect the onset of diseases such as Alzheimer’s disease, Parkinson’s disease, and diabetes , 2017, Free radical research.

[24]  M. Zaiss,et al.  The double-edged role of 12/15-lipoxygenase during inflammation and immunity. , 2017, Biochimica et biophysica acta. Molecular and cell biology of lipids.

[25]  Olivier Michielin,et al.  SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules , 2017, Scientific Reports.

[26]  K. Rahman,et al.  Aged garlic has more potent antiglycation and antioxidant properties compared to fresh garlic extract in vitro , 2017, Scientific Reports.

[27]  A. Pisoschi,et al.  Antioxidant Capacity Determination in Plants and Plant-Derived Products: A Review , 2016, Oxidative medicine and cellular longevity.

[28]  S. Afrin,et al.  IN VITRO ANTIOXIDANT ACTIVITY, ANTIMICROBIAL AND PRELIMINARY CYTOTOXIC ACTIVITY OF CYNOMETRA RAMIFLORA- A MANGROVE PLANT , 2016 .

[29]  Yukun Yang,et al.  Isolation, purification and identification of antioxidants in an aqueous aged garlic extract. , 2015, Food chemistry.

[30]  Fereidoon Shahidi,et al.  Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects – A review , 2015 .

[31]  Douglas E. V. Pires,et al.  pkCSM: Predicting Small-Molecule Pharmacokinetic and Toxicity Properties Using Graph-Based Signatures , 2015, Journal of medicinal chemistry.

[32]  H. Kuhn,et al.  Mammalian lipoxygenases and their biological relevance. , 2015, Biochimica et biophysica acta.

[33]  V. Lushchak.  Free radicals, reactive oxygen species, oxidative stress and its classification. , 2014, Chemico-biological interactions.

[34]  C. Dani,et al.  Antioxidant Strategies and Respiratory Disease of the Preterm Newborn: An Update , 2014, Oxidative medicine and cellular longevity.

[35]  Eunjung Lee,et al.  Anti-inflammatory activity of rhamnetin and a model of its binding to c-Jun NH2-terminal kinase 1 and p38 MAPK. , 2014, Journal of natural products.

[36]  O. Ogbu,et al.  Phytochemical and Antioxidant Analyses of Selected Edible Mushrooms, Ginger and Garlic from Ebonyi State, Nigeria. , 2014 .

[37]  Shuxia Chen,et al.  Evaluation of Garlic Cultivars for Polyphenolic Content and Antioxidant Properties , 2013, PloS one.

[38]  Atta-ur-rahman,et al.  An investigation of phenolic compounds from plant sources as trypsin inhibitors , 2012, Natural product research.

[39]  Hafiz Ansar Rasul Suleria,et al.  Aqueous garlic extract and its phytochemical profile; special reference to antioxidant status , 2012, International journal of food sciences and nutrition.

[40]  J. Agbedahunsi,et al.  In vitro evaluation of membrane stabilizing activities of leaf and root extracts of Calliandra portoricensis (JACQ) benth on sickle and normal human erythrocytes. , 2012 .

[41]  I. T. Ten Berge,et al.  Serine proteases of the human immune system in health and disease. , 2010, Molecular immunology.

[42]  R. Bushra,et al.  An overview of clinical pharmacology of Ibuprofen. , 2010, Oman medical journal.

[43]  G. Goncagul,et al.  Antimicrobial effect of garlic (Allium sativum). , 2010, Recent patents on anti-infective drug discovery.

[44]  G. Sampaio,et al.  Garlic (Allium sativum L.) and ready-to-eat garlic products: In vitro antioxidant activity , 2009 .

[45]  L. Włodek,et al.  Biological properties of garlic and garlic‐derived organosulfur compounds , 2009, Environmental and molecular mutagenesis.

[46]  N. Turhan,et al.  Hepatic oxidant/antioxidant status in cholesterol‐fed rabbits: Effects of garlic extract , 2009, Hepatology research : the official journal of the Japan Society of Hepatology.

[47]  N. Mimica-Dukić,et al.  Phenolics as antioxidants in garlic (Allium sativum L., Alliaceae) , 2008 .

[48]  K. Arai,et al.  Novel lipoxygenase inhibitors as neuroprotective reagents , 2008, Journal of neuroscience research.

[49]  C. Funk.  Lipoxygenase pathways as mediators of early inflammatory events in atherosclerosis. , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[50]  Mi-Yeon Kim,et al.  Identification and in vitro biological activities of flavonols in garlic leaf and shoot: inhibition of soybean lipoxygenase and hyaluronidase activities and scavenging of free radicals , 2005 .

[51]  B. Yeğen,et al.  Aqueous garlic extract alleviates ischaemia‐reperfusion‐induced oxidative hepatic injury in rats , 2005, The Journal of pharmacy and pharmacology.

[52]  K. E. Malterud,et al.  Antioxidant activity in extracts from coriander , 2004 .

[53]  Conrad C. Huang,et al.  UCSF Chimera—A visualization system for exploratory research and analysis , 2004, J. Comput. Chem..

[54]  B. Lau,et al.  Aged Garlic Extract and its Constituents Inhibit Cu2+-Induced Oxidative Modification of Low Density Lipoprotein , 1997, Planta medica.

[55]  C. Berset,et al.  Use of a Free Radical Method to Evaluate Antioxidant Activity , 1995 .

[56]  I. Popov,et al.  Antioxidant effects of aqueous garlic extract. 2nd communication: Inhibition of the Cu(2+)-initiated oxidation of low density lipoproteins. , 1994, Arzneimittel-Forschung.

[57]  M. Kunitz,et al.  CRYSTALLINE SOYBEAN TRYPSIN INHIBITOR : II. GENERAL PROPERTIES. , 1947 .

[58]  Molecular Optimization, Docking and Dynamic Simulation Study of Selective Natural Aromatic Components to Block E2-CD81 Complex Formation in Predating Protease Inhibitor Resistant HCV Influx , 2022, International Journal of Pharmaceutical Research.