Inhibitory Potential of Pulutan (<i>Urena lobata</i>) Leaf Extract on Inducible Nitric Oxide Synthase as Anti-inflammatory Agent: <i>In Vitro</i> and <i>In Silico</i> Approaches
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Abstract
Urena lobata is an herbal remedy traditionally used to cure certain ailments. Its flavonoid and phenol-rich content has been shown to have free radical scavenging activity. Nitric oxide (NO) is an active gaseous substance that is released by cells and plays a role in inflammation. This study examined the ability of U. lobata leaf extract to induce NO production and suppress inducible NO synthase (iNOS). The leaves of U. lobata were extracted with ethanol. The phytoconstituents of the extract were identified, their physicochemical and pharmacokinetic properties were predicted, and their anti-inflammatory activity was evaluated. The structures of the phytoconstituents were determined using the molecular docking approach. Griess technique was used to evaluate NO generation in IL-1β-treated hepatocyte culture media containing the extract. An in silico approach was employed to investigate the suppression of iNOS by phytoconstituents from U. lobata. The results showed that stigmasterol and gossypetin were identified in the extract and had a strong inhibitory effect on iNOS based on the inhibition constant. However, it has lower potency than β-sitosterol. The ethanol extract of U. lobata (IC50 = 163.03 µg/mL) had a lower potency than gossypin (IC50 = 100.31 µg/mL) but was comparable to gossypetin (IC50 = 164.18 µg/mL) for inhibiting NO generation. The ethanol extract of U. lobata demonstrated anti-inflammatory action by lowering NO level, but at a lower concentration than gossypin and gossypetin. The study revealed that U. lobata ethanol extract inhibited NO production through active substances, like gossypetin, mangiferin, β-sitosterol, and stigmasterol, thereby suppressing iNOS activity.
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References
Babu SS, Madhuri DB, Ali SL. A pharmacological review of Urena lobata plant. Asian J Pharm Clin Res. 2016; 9(2): 20-22.
Pusvita S, Rohama, Yuwindry I, Darsono PV. Antibacterial Activity Test of Pulutan Leaf Extract (Urena lobata L) Against Streptococcus pyogenes Bacteria. J Pharm Care Sci. 2023; 4(1):34-41.
Garuba T, Katrodiya N, Patel N, Patel S, Rajani DP, Chettiar SS, Abdulrahman AA, Krishnamurthy R. Antibacterial activity of Urena lobata against uropathogens. Nig J Nat Prod Med. 2021; 25(1): 43-46.
Hassanpour SH, Doroudi A. Review of the antioxidant potential of flavonoids as a subgroup of polyphenols and partial substitute for synthetic antioxidants. Avicenna J Phytomed. 2023;13(4):354-376.
Purnomo Y, Soeatmadji DW, Sumitro SB, Widodo MA. Inhibitory activity of Urena lobata leaf extract on dipeptidyl peptidase-4 (DPP-4): is it different in vitro and in vivo?. Med Plants. 2018; 10(2):99-105.
Evans WC. Trease and Evans Pharmacognosy (15th edition). India: Elsevier; 2002.
Al-Khayri JM, Sahana GR, Nagella P, Joseph BV, Alesa FM, Al-Mssallem MQ. Flavonoids as potential anti-inflammatory molecules: A review. Molecules. 2022; 27(9): 2901.
Zhao Q, Zhu L, Wang S, Gao Y, Jin F. Molecular mechanism of the anti-inflammatory effects of plant essential oils: A systematic review. J Ethnopharmacol. 2023; 301: 115829.
Król M, Kupnicka P, Bosiacki M, Chlubek, D. Mechanisms underlying anti-inflammatory and anti-cancer properties of stretching—a review. Int J Mol Sci. 2022; 23(17): 10127.
Zhao XY, Lempke SL, Urbán Arroyo JC, Yin B, Holness NK, Smiley J, Ewald SE. Inducible nitric oxide synthase (iNOS) is necessary for GBP-mediated T. gondii restriction in murine macrophages via vacuole nitration and intravacuolar network collapse. bioRxiv. 2023.
Chen L, Deng H, Cui H, Fang J, Zuo Z, Deng J, Li Y, Wang X, Zhao L. Inflammatory responses and inflammation-associated diseases in organs. Oncotarget. 2017; 9(6): 7204-7218.
Ningsih FN, Okuyama T, To S, Nishidono Y, Okumura T, Tanaka K, Ikeya Y, Nishizawa, M. Comparative analysis of anti-inflammatory activity of the constituents of the rhizome of Cnidium officinale using rat hepatocytes. Bio Pham Bull. 2020; 43(12):1867-1875.
Kanemaki T, Kitade H, Hiramatsu Y, Kamiyama Y, Okumura T. Stimulation of glycogen degradation by prostaglandin E2 in primary cultured rat hepatocytes. Prostaglandins. 1993; 45: 459–474.
Green LC, Wagner DA, Glogowski J, Skipper Pl, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite and nitrate in biological fluids. Anal Biochem, 1982; 126:131–138.
Ohno T, Morita H, Arae K, Matsumoto K, Nakae S. Interleukin‐33 in allergy. Allergy. 2012; 67(10), 1203-1214.
Liu T, Zhang L, Joo D, Sun SC. NF-κB signaling in inflammation. Sig Transduct Target Ther. 2017;2:17023
Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev. 1997; 23(1-3):3-26.
Naglah AM, Askar AA, Hassan AS, Khatab TK, Al-Omar MA, Bhat MA. Biological evaluation and molecular docking with in silico physicochemical pharmacokinetic and toxicity prediction of pyrazolo[15-a] pyrimidines. Molecules. 2020; 25(6):1431.
Adianingsih OR, Khasanah U, Anandhy KD, Yurina V. In silico ADME-T and molecular docking study of phytoconstituents from Tithonia diversifolia (Hemsl.) A. Gray on various targets of diabetic nephropathy. J Pharm Pharmacogn Res. 2022; 10(4):571-594.
Kenny PW. Hydrogen-bond donors in drug design. J Med Chem. 2022; 65.21:14261-14275.
Daina A, Michielin O, And Zoete V. SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Scientific reports. 2017; 7(1):42717.
Ndombera FT, Maiyoh GKK, Vivian CT. Pharmacokinetic, Physicochemical and Medicinal Properties of N-Glycoside Page 2 of 8 Anti-Cancer Agent more Potent than 2-Deoxy-D-Glucose in Lung Cancer Cells. Cancer Sci Res, 2019; 6(1):1-8.
Ahmad I, Kuznetsov AE, Pirzada AS, Alsharif KF, Daglia M, And Khan H. Computational pharmacology and computational chemistry of 4-hydroxyisoleucine: Physicochemical, pharmacokinetic, and DFT-based approaches. Front Chem. 2023; 11:1145974.
Cournia Z, Allen B, Sherman W. Relative binding free energy calculations in drug discovery: recent advances and practical considerations. J Chem Inf Model. 2017; 57(12):2911-2937.
Doganyigit Z, Eroglu E, Akyuz E. Inflammatory mediators of cytokines and chemokines in sepsis: From bench to bedside. Hum Exp Toxicol. 2022; 41.
Agu PC, Afiukwa CA, Orji OU, Ezeh EM, Ofoke IH, Ogbu CO, Ugwuja EI, Aja PM. Molecular docking as a tool for the discovery of molecular targets of nutraceuticals in diseases management. Sci Rep. 2023; 13(1):13398.
Meli R, Morris GM, Biggin PC. Scoring Functions for Protein-Ligand Binding Affinity Prediction using Structure-Based Deep Learning: A Review. Front Bioinform. 2022; 2:885983.
Sourabh S. Baghel. Sonal D. Prashant S. Priya S. Yogesh S. Acute Toxicity Study of Aqueous Extract of Coccinia indica (Roots). Asian J Pharm Sci. 2011; 1(1):23-25.
Rajčević N, Bukvički D, Dodoš T, Marin PD. Interactions between natural products—A review. Metabolites. 2022; 12(12); 1256.
Purnomo Y, Tilaqza A. Analgesic and Anti-inflammatory Activities of Urena lobata L. Leaf Extracts. Indonesian J Pharm. 2022; 33(4):566-574.
Purnomo Y, Wahyunigsih D, Tilaqza A. Anti-inflammatory Potency of Pulutan (Urena lobata) Leaf extract and its Fractions by Protein Denaturation Inhibition Assay. Res J Pharm Techno. 2023; 16(11):5406 – 5409.
Purnomo Y, Aini N, Noerhayati E. Acute Toxicity Level of Pulutan (Urena lobata) Leaf Extract on Zebrafish (Danio rerio) and its Analysis by In Silico Study. Res J Pharm Techno. 2022; 15(6):2477-2482.