Dechlorophyllized Urena lobata Leaf Fraction Inhibits Nitric Oxide Production in IL-1β-Induced Rat Hepatocytes
Article Sidebar
Main Article Content
Abstract
Urena lobata has demonstrated anti-inflammatory properties due to its rich flavonoid content. Chlorophyll, a key anti-inflammatory pigment, can inhibit pure compound isolation, necessitating chlorophyll removal to improve quantification and pharmacological activity. The present study was conducted to evaluate the effect of dechlorophyllized ethyl acetate (EtOAc) fraction of U. lobata on nitric oxide (NO) production in interleukin (IL)-1β-induced rat hepatocytes. The ethanol extract of U. lobata leaf fraction was fractionated using EtOAc solvent, some dechlorophyllized, and identified using liquid chromatography for phytoconstituent targets. Primary rat hepatocytes were isolated and cultured, then treated with U. lobata leaf fraction (25–100 µg/mL) and the dechlorophyllized fraction (50–200 µg/mL) in the presence of IL-1β for 8 hours. Nitric oxide levels were measured using the Griess method, and IC50 values were determined. The results showed that the dechlorophyllized U. lobata leaf fraction increased NO levels by approximately 70% compared to the fraction without chlorophyll removal. However, the U. lobata leaf fraction (IC50 = 78.1 ppm) exhibited stronger NO inhibition than the dechlorophyllized fraction (IC50 = 129.77 ppm). The intensity value of gossypin (GPI) decreased approximately threefold in the dechlorophyllized U. lobata fraction (25 mAU) compared to the fraction without chlorophyll removal (75 mAU). The dechlorophyllized U. lobata leaf fraction also exhibited reduced NO inhibitory activity and a lower GPI level as a phytoconstituent. These findings emphasize the role of chlorophyll and associated phytoconstituents in the pharmacological activity of U. lobata and the need for further toxicity assessments to ensure its safety.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
1. Purnomo Y, Tilaqza A. Analgesic and anti-inflammatory activities of Urena lobata leaf extract. Indonesian J Pharm. 2022; 33(4):566-574.
2. Islam MH, Rahman KMH, Rahman S, Rahmatullah M. Preliminary antihyperglycemic, anti-nociceptive activity, phytochemical analysis and toxicity studies on leaves of Urena lobata L. J Chem Pharm Res. 2015; 7(4):559-563.
3. Purnomo Y, Makdasari J, Fatahillah FI. Inhibitory activity of Urena lobata leaf extract on alpha-amylase and alpha-glucosidase: in vitro and in silico approach. J Basic Clin Physiol Pharmacol. 2021; 32(4):889-894.
4. Kanimozhi D, Bai VR. Evaluation of phytochemical antioxidant, antimicrobial activity, determination of bioactive components of ethanolic extract of aerial and underground parts of Cynodon dactylon L. Int J Sci Res Rev. 2012; 1:33-48.
5. Adewale AO, David AA, Abiodun OO. Studies on antimicrobial, antioxidant and phytochemical analysis of Urena lobata leaf extract. J Phys Nat Sci. 2007; 1(2):12-20.
6. Wahyuningsih D, Purnomo Y, Tilaqza A. In silico study of Pulutan (Urena lobata) leaf extracts as anti-inflammatory agents and their prediction. J Trop Pharm Chem. 2022; 1(6):30-37.
7. Kunnumakkara AB, Nair AS, Ahn KS, Pandey MK, Yi Z, Liu M, Aggarwal BB. Gossypin, a pentahydroxy glucosyl flavone, inhibits the transforming growth factor beta-activated kinase-1-mediated NF-kappaB activation pathway, leading to potentiation of apoptosis, suppression of invasion, and abrogation of osteoclastogenesis. Blood. 2007; 109(12):5112-5121. https://doi.org/10.1182/blood-2007-01-067256.
8. Purnomo Y, Tilaqza A. Inhibitory potential of Pulutan (Urena lobata) leaf extract on inducible nitric oxide synthase as anti-inflammatory agent: in vitro and in silico approaches. Trop J Nat Prod Res. 2024; 8(8):8195-8201.
9. Singh AK, Rana HK, Panday AK. Analysis of chlorophylls. In: Recent Advances in Natural Products Analysis. Elsevier: 2020; 635-650. https://doi.org/10.1016/B978-0-12-816455-6.00019-6
10. Tzima K, Brunton NP, Rai DK. Evaluation of the impact of chlorophyll removal techniques on polyphenols in rosemary and thyme by-products. J Food Biochem. 2020; 1-22.
11. Kim SB, Bisson J, Friesen JB, Pauli GF, Simmler C. Selective chlorophyll removal method to "degreen" botanical extracts. J Nat Prod. 2020; 83(6):1846-1858.
12. Flieger J, Zuk N, Patkowska SP, Kusmierz M, Panek R, Franus W, Baj J, Buszewicz G, Terensinski G, Plazinski W. Selective removal of chlorophyll and isolation of lutein from plant extracts using magnetic solid-phase extraction with iron oxide nanoparticles. Int J Mol Sci. 2024; 25(3152):1-22.
13. Colasanti M, Suzuki H. The dual personality of NO. Trends Pharmacol Sci. 2000; 21:249-252.
14. Yoshigai E, Machida T, Okuyama T, Mori M, Murase H, Yamanishi R, Okumura T, Ikeya Y, Nishino H, Nishizawa M. Citrus nobiletin
suppresses inducible nitric oxide synthase gene expression in interleukin-1β-treated hepatocytes. Biochem Biophys Res Commun. 2013; 439(1):54-59.
15. Cotran RS, Kumar V, Collins T, Robbins SL. Robbins Pathologic Basis of Diseases. 6th ed. Philadelphia: WB Saunders Company; 2002:12-21.
16. Kitade H, Sakitani K, Inoue K, Masu Y, Kawada N, Hiramatsu Y, Kamiyama Y, Okumura T, Ito S. Interleukin-1β markedly stimulates nitric oxide formation in the absence of other cytokines or lipopolysaccharide in primary cultured rat hepatocytes but not in Kupffer cells. Hepatology. 1996; 23:797-802.
17. 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.
18. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, et al. Analysis of nitrate, nitrite, and nitrate in biological fluids. Anal Biochem. 1982; 126:131-138.
19. Ohno N, Yoshigai E, Okuyama T, Yamamoto Y, Okumura T, et al. Chlorogenic acid from the Japanese herbal medicine Kinginka (Flos Lonicerae japonicae) suppresses the expression of inducible nitric oxide synthase in rat hepatocytes. HOAJ Biol. 2012; 1:2.
20. Dwijayanti DR, Shimada T, Ishii T, Okuyama T, Ikeya Y, Mukai E, Nishizawa M. Bitter melon fruit extract has a hypoglycemic effect and reduces hepatic lipid accumulation in ob/ob mice. Phytother Res. 2019; 1-9.
21. Evans WC. Trease and Evans Pharmacognosy. 15th ed. Elsevier, A Division of Reed Elsevier India Pvt. Limited; 2002.
22. Brunton LL, Lazo JS, Parker KL. Goodman & Gilman's the Pharmacological Basis of Therapeutics. New York: McGraw-Hill; 2006.
23. 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.
24. Szakács G, Váradi A, Ozvegy-Laczka C, Sarkadi B. The role of ABC transporters in drug absorption, distribution, metabolism, excretion, and toxicity (ADME-Tox). Drug Discov Today. 2008; 13:379-393.
25. Tiwari P, Kumar B, Kaur B, Kaur G, Kau H. Phytochemical screening and extraction: A review. Int Pharm Sci. 2011; 1(1):98-106.
26. Purnomo Y, Aini N, Noerhayati E. Acute toxicity level of Pulutan (Urena lobata) leaf extract on Danio rerio and its analysis by in silico study. Res J Pharm Tech. 2022; 15(6):2477-2482.
27. Omonkhua AA, Onoagbe IO. Evaluation of the long-term effect of Urena lobata root extracts on blood glucose and hepatic function of normal rabbits. J Toxicol Environ Health. 2011; 209:1-8. doi.org/10.5897/JTEHS.9000031