In silico, in-vitro and in vivo screening of biological activities of citral
Abstract
Abstract. Citral, one of the main components of lemongrass oil (65–85%), is known to possess various medicinal properties like enhancing skin health and vision-improvement. It also acts as flavoring agent, used in perfumes and skin care products. The objective of this work was to elucidate the biological properties of citral at molecular level using an integrated in silico, in vitro and in vivo approaches. To elucidate this in silico molecular docking studies were performed with in vitro validation by DPPH scavenging activity, MTT assays, enzymatic assays and Chorio Allantoic Membrane (CAM) assay. The in silico analysis demonstrated the potential binding of citral with PPARγ ligand binding domain and vascular endothelial growth factor receptors (VEGFR-1 and VEGFR-2). Citral is already a proven anti-oxidant which is further confirmed by increased DPPH inhibition with increased citral concentration (IC50: 6.9 ± 1.68 μg/ml, p < 0.05). The results demonstrated that citral protect yeast cells from cytotoxic effects of hydrogen peroxide and also increase the activities of antioxidant enzymes like GST, SOD and LPO. It was also demonstrated to be cytotoxic to cancerous HeLa cells (IC50: 3.9 ± 0.38 μM, p < 0.01) and was found anti-angiogenic by CAM assay. This study highlights many important pharmaceutical properties of citral which can be explored further to increase its industrial applications.
References
1 . The traditional medicine and modern medicine from natural products. Molecules. 2016;21(5):559.
2 . Citral: Identifying a threshold for induction of dermal sensitization. Regul. Toxicol. Pharm. 2008;52(1):62–73.
3 . Beverage emulsions: Recent developments in formulation, production, and applications. Food Hydrocoll. 2014;42:5–41.
4 . Pharmacological study of Cymbopogon citratus leaves. J Ethnopharmacol. 1989;25(1):103–107.
5 . Lemongrass and citral effect on cytokines production by murine macrophages. J Ethnopharmacol. 2011;137(1):909–913.
6 . Inactivation of Escherichia coli by citral. J Appl Microbiol. 2010;108(6):1928–1939.
7 Antimicrobial activity and possible mechanism of action of citral against Cronobacter sakazakii. PLoS One. 2016;11(7):e0159006.
8 . Antifungal activity of the lemongrass oil and citral against Candida spp. Braz J Infect Dis. 2008;12(1):63–66.
9 . Antimicrobial, antioxidant, and antitumor activity of epsilon-poly-L-lysine and citral, alone or in combination. Food Nutr Res. 2016;60(1):31891.
10 . Antipyretic effects of citral and possible mechanisms of action. Inflammation. 2017;40(5):1735–1741.
11 . The biology of peroxisome proliferator-activated receptors: relationship with lipid metabolism and insulin sensitivity. Diabetes. 2004;53(suppl 1):43–50.
12 . Nuclear receptors of the peroxisome proliferator-activated receptor (PPAR) family in gestational diabetes: from animal models to clinical trials. Biol Reprod. 2010;83(2):168–176.
13 . Citral, a component of lemongrass oil, activates PPARα and γ and suppresses COX-2 expression. Biochim Biophys Acta. 2010;1801(11):1214–1220.
14 . Citral inhibits lipopolysaccharide-induced acute lung injury by activating PPAR-γ. Eur J Pharmacol. 2015;747:45–51.
15 . AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem. 2010;31(2):455–461.
16 . Antioxidant and free radical-scavenging activity of Choto-san and its related constituents. Biol Pharm Bull. 2004;27(1):38–46.
17 . Use of MTT colorimetric assay to measure cell activation. J Immunol Methods. 1986;94(1–2):57–63.
18 . Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry. 1971;44(1):276–287.
19 . Glutathione S-transferases the first enzymatic step in mercapturic acid formation. J Biol Chem. 1974;249(22):7130–7139.
20 . Glutathione metabolizing enzymes and oxidative stress in ferric nitrilotriacetate mediated hepatic injury. Redox Rep. 1996;2(6):385–391.
21 . Chick chorioallantoic membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. Int J Mol Sci. 2012;13(8):9959–9970.
22 . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–275.
23 . Essential oils, part V: peppermint oil, lavender oil, and lemongrass oil. Dermatitis. 2016;27(6):325–332.
24 . Comparison of extraction methods for marker compounds in the essential oil of lemon grass by GC. J Agric Food Chem. 2002;50(6):1345–1349.
25 . Pharmacological study of Cymbopogon citratus leaves. J Ethnopharmacol. 1989;25(1):103–107.
26 . Structural insight into PPARγ ligands binding. Curr Med Chem. 2009;16(14):1768–1789.
27 . Lemon grass (Cymbopogon citratus) essential oil as a potent anti-inflammatory and antifungal drugs. Libyan J Med. 2014;9(1):25431.
28 Essential oils from Ugandan medicinal plants: in vitro cytotoxicity and effects on IL-1β-induced proinflammatory mediators by human gingival fibroblasts. Evid Based Complement. Alternat Med. 2016;2016.
29 . Lemongrass effects on IL-1β and IL-6 production by macrophages. Nat Prod Res. 2009;23(12):1151–1159.
30 . Inhibitory effect of citral on NO production by suppression of iNOS expression and NF-κB activation in RAW264. 7 cells. Arch Pharm Res. 2008;31(3):342–349.
31 Effect of a glutathione S-transferase inhibitor on oxidative stress and ischemia-reperfusion-induced apoptotic signalling of cultured cardiomyocytes. Exp Clin Cardiol. 2011;16(3):92.
32 . Biological properties of citral and its potential protective effects against cytotoxicity caused by aspirin in the IEC-6 cells. Biomed Pharmacother. 2017;87:653–660.
33 Anticancer activity of an essential oil from Cymbopogon flexuosus. Chem Biol Interact. 2009;179(2–3):160–168.
34 . Compositional Variability in Leaves and Inflorescence Essential Oils of Cymbopogon khasianus (Hack.) Stapf ex Bor Collected from Meghalaya: A Biodiversity Hotspot. Journal of Essential Oil Bearing Plants. 2018;21(3):640–657.
35 . Assessment of Selection Criteria for Development of High Yielding Genoptypes Using Variability Parameters in Lemongrass Cymbopogon flexuosus L. Journal of Essential Oil Bearing Plants. 2017;20(6):1450–1460.
36 . Preformed antifungal compounds of lemon fruit: citral and its relation to disease resistance. J Agric Food Chem. 1995;43(4):1057–1061.
37 Effect of citral on mouse hepatic cytochrome P450 enzymes. Pharm Biol. 2018;56(1):337–343.
38 Hepatoprotective effect of citral on acetaminophen-induced liver toxicity in mice. Evid Based Complement Alternat Med. 2017;2017.