nps Natural Product Sciences Natural Product Sciences 1226-3907 2288-9027 The Korean Society of Pharmacognosy 10.20307/nps.2016.22.2.122 nps-22-122 Original Article Cytotoxic, Anti-Inflammatory and Adipogenic Effects of Inophyllum D, Calanone, Isocordato-oblongic acid, and Morelloflavone on Cell Lines Taher Muhammad 1 ∗ Aminuddin Amnani 2 Susanti Deny 3 Aminudin Nurul Iman 4 On Shamsul 4 Ahmad Farediah 4 Hamidon Hanisuhana 1 Department of Pharmaceutical Technology, Faculty of Pharmacy, International Islamic University Malaysia, Jalan Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Pahang, Malaysia Department of Biomedical Science, Faculty of Science, International Islamic University Malaysia, Jalan Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Pahang, Malaysia Department of Chemistry, Faculty of Science, International Islamic University Malaysia, Jalan Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Pahang, Malaysia Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor, Malaysia Author for correspondence Muhammad Taher, Department of Pharmaceutical Technology, Faculty of Pharmacy, International Islamic University Malaysia, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia Tel: +60-95704807; E-mail: mtaher@iium.edu.my 122016 17122016 222 122128 23112015 21012016 24012016 Copyright © 2016 The Korean Society of Pharmacognosy 2016 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract

This paper reports in vitro cytotoxic, anti-inflammatory and adipocyte diffentiation with adipogenic effects of coumarins inophyllum D (1) and calanone (2), and a chromanone carboxylic acid namely isocordato-oblongic acid (3) isolated from Calophyllum symingtonianum as well as a biflavonoid morelloflavone (4) isolated from Garcinia prainiana on MCF-7 breast adenocarcinoma RAW 264.7 macrophages and 3T3-L1 preadipocytes cells, respectively. The cytotoxicity study on MCF-7 cell was conducted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Meanwhile, the study of anti-inflammatory effects in RAW 264.7 macrophages and adipogenic effects on 3T3-L1 pre-adipocytes were conducted through nitrite determination assay and induction of adipocyte differentiation, respectively. In the cytotoxicity study, inophyllum D (1) was the only compounds that exhibited significant cytotoxic effect against MCF-7 cell with IC50 of 84 µg/mL. Further, all by inhibiting the compounds have shown anti-inflammatory effects in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages of nitrite concentration with production. In addition, the compounds also exhibited adipogenic effects on 3T3-L1 pre-adipocytes by stimulating lipid formation. Thus, this study may provide significant input in discovery of the potential effects cytotoxic, anti-inflammatory and adipogenic agents.

 Cytotoxic Anti-inflammatory Adipogenic REFERENCES Hanson J. R. Royal Society of Chemistry 2003 17 Croteau R. Kutchan T. M.; Lewis, N.G. Biochem. Mol. Biol. Plants 2000 24 12501319 Brahmachari G. Bioactive Natural Products; John Wiley & Sons: United Kingdom 2015 1199 Aminudin N. I. Ahmad F. Taher M. Zulkifli R. M. Nat. Prod. Commun. 2015 10 15851587 Waterman P. G. Crichton E. G. Phytochemistry 1980 19 27232726 Mosmann T. J. Immunol. Methods 1983 65 5563 Lee H. S. Ryu D. S. Lee G. S. Lee D. S. J. Ethnopharmacol. 2012 140 271276 Susanti D. Amiroudine M. Z. Rezali M. F. Taher M. Nat. Prod. Res. 2013 27 417424 Itoigawa M. Ito C. Tan H. T. Kuchide M. Tokuda H. Nishino H. Furukawa H. Cancer Lett. 2001 169 1519 Osman A. M. Bayoumi H. M. Al-Harthi S. E. Damanhouri Z. A. ElShal M. F. Cancer Cell Int. 2012 12 4753 Gao Y. Jiang W. Dong C. Li C. Fu X. Min L. Tian J. Jin H. Shen J. Toxicol. In Vitro 2012 26 1 16 Nahar P. P. Driscoll M. V. Li L. Slitt A. L. Seeram N. P. J. Funct. Foods 2014 6 126136 Kumar V. Abbas A. K. Fausto N. Mitchell R. Robbins Basic Pathology: 8th Edition; Saunders/Elsevier: Philadelphia 2007 9 Kim H. K. Cheon B. S. Kim Y. H. Kim S. Y. Kim H. P. Biochem. Pharmacol. 1999 58 759765 Macia L. Viltart O. Verwaerde C. Delacre M. Delanoye A. Grangette C. Wolowczuk I. Genes Nutr. 2006 1 189212 Yun J. W. Phytochemistry 2010 71 16251641 Figures

Structure of tested compounds.

Cytotoxic effects of selected flavonoids on MCF-7 cells by MTT assay. Results are presented as mean ± SD of triplicate of three experiments. (A) Calanone (B) Morelloflavone (C) Isocordato-oblongic acid (D) Inophyllum D.

Cytotoxic effects of selected flavonoids on RAW 264.7 cells by MTT assay. Results are presented as mean ± SD of triplicate of three experiments.

Anti-inflammatory effects of selected flavonoids on RAW 264.7 cells by nitrite determination test. The data shown were nitrite concentration against experimental groups for (A) Calanone (B) Morelloflavone (C) Isocordato-oblongic Acid (D) Inophyllum D. Results are presented as mean ± SD of triplicate from one independent experiment.

Cytotoxic effects of selected flavonoids on 3T3 L1 pre-adipocyte. Results are presented as mean ± SD of triplicates of two repeated experiments.

Effects of selected flavonoids on 3T3-L1 differentiation. All values were presented as mean ± SD indicating a measure of the dispersion of a set of data from its means from one independent experiment with three replications.

Adipogenic effect of inophyllum D (A), calanone (B), morelloflavone (C) and isocordato-oblogic acid (D) at 25 µ g/mL on 3T3-L1 adipocyte. The cells were stained with Oil red O at 10 days after induction (400× magnification). Arrows indicated the presence of lipid droplet.