The Effects of Mulberry Leaves (Morus alba L.) Extract on Lipid Accumulation in Adipocytes 3T3-L1 Cells and in Vitro Cytotoxic Activity
Article Sidebar
Main Article Content
Abstract
Diabetes mellitus is a disease that occurs when the pancreas cannot produce enough insulin or when the body cannot effectively use the insulin that is produced. Obesity is a major risk factor for diabetes, characterised by excessive lipid accumulation in adipose tissue. This study aims to determine the effects of a 70% ethanol extract from mulberry leaves (Morus alba L.) and its fractions on inhibiting lipid accumulation in 3T3-L1 adipocyte cell lines (preadipocytes derived from mouse embryonic fibroblasts). Mulberry leaves were macerated with 70% ethanol and partitioned with n-hexane and ethyl acetate. The extract of mulberry leaves and its fractions were first tested for cytotoxicity using the MTT assay method on 3T3-L1 cells. The 3T3-L1 cells were differentiated using differentiation agents. After differentiation, the cells were treated with samples of ethanol extract and its fractions at concentrations of 31.25, 62.5, 125, 250, 500, and 1000 ppm. The IC50 values obtained were 416 µg/mL for the ethanol extract, 49 µg/mL for the n-hexane fraction, 89 µg/mL for the ethyl acetate fraction, and 225 µg/mL for the water fraction. The results of the lipid accumulation test show that the ethyl acetate, n-hexane, and water fractions reduce lipid accumulation, but the ethanol extract increases lipid accumulation. According to these findings, the ethanol extract fractions from mulberry leaves can inhibit lipid accumulation in 3T3L1 adipocyte cells.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
References
Chikara S, Nagaprashantha, LD, Singhal J, Horne D, Awasthi S, Singhal S. Oxidative stress and dietary phytochemicals: Role in cancer chemoprevention and treatment. Cancer Lett. 2018;413,122-134.
Das PR, Darwish A G , Ismail A, Haikal AM., Gajjar P, Balasubramani SP, Sheikh MB, Tsolova V, Soliman KF, Sherif SM. Diversity in blueberry genotypes and developmental stages enables discrepancy in the bioactive compounds, metabolites, and cytotoxicity. Food Chem. 2022;374, 131632.
Dat NT, Binh PTX., Van MC, Huong HT, Lee, JJ. Cytotoxic prenylated flavonoids from Morus alba. Fitoterapia, 2010;81(8),1224-1227.
Deepa M, Sureshkumar T, Satheeshkumar PK, Priya S. Antioxidant rich Morus alba Leaf extract induces apoptosis in human colon and breast cancer cells by the downregulation of nitric oxide produced by inducible nitric oxide synthase. Nutr Cancer. 2013; 65(2), 305-310.
Deka SJ, Gorai S, Manna D, Trivedi, V. Evidence of PKC binding and translocation to explain the anticancer mechanism of chlorogenic acid in breast cancer cells. Curr. Mol. Med. 2017;17(1), 79-89.
Hyun TK, Kim MO, Lee H, Kim Y, Kim E, Kim JS. Evaluation of antioxidant and anticancer properties of Dendropanax morbifera Léveille. Food Chem. 2013; 141(3), 1947-1955.
Izzreen MNQ, Fadzelly AM, Hartina MU, Amirah RR, Rozzamri, A. A cytotoxicity and sub-acute toxicity study on tea leaves cultivated in Sabah. Int. Food Res. J. 2020;27(5), 925-933.
Jelled A, Hassine RB, Thouri A, Flamini G, Chahdoura, H, Arem A, Lamine JB, Haouas Z, Cheikh HB, Achour, L. Immature mulberry fruits richness of promising constituents in contrast with mature ones: A comparative study among three Tunisian species. Ind Crops Prod. 2017;95, 434-443.
Themelis T, Boschetti S, Gelli M, Gatti R. Simple Determination of 1- Deoxynojirimycin in a New Dietary Supplement by Liquid Chromatography. Chromatographia. 2018; 81(11), 1495-1501.
Wulandari YRE, Prasasty VD, Rio A. and Geniola, C. Determination of 1-Deoxynojirimycin Content and Phytochemical Profiles from Young and Mature Mulberry Leaves of Morus spp. J. Biol. Sci. 2019;19(2), 124-131.
Metwally FM, Rashad H, Mahmoud AA. Morus alba L diminishes visceral adiposity, insulin resistance, behavioral altera- tions via regulation of gene expression of leptin, resistin and adiponectin in rats fed a high-cholesterol diet. Physiol. Behav. 2019; 201, 1–11.
Shin NR, Bose S, Choi Y, Kim YM, Chin YW, Song EJ, Nam YD, Kim H. Anti-obesity effect of fermented panax notoginseng is mediated via modulation of appetite and gut microbial population. Front. Pharmacol. 2021;12, 665881.
Yu MH, Tsai MC, Wang CC, Wu SW, Chang YJ, Hsun CS, Wang CJ. Mulberry leaf polyphenol extract and rutin induces autophagy regulated by p53 in human hepatoma HepG2 cells. Pharmaceuticals (Basel). 2021; 8(12); 234-247.
Deepa M, Sureshkumar T, Satheeshkumar PK, Priya S. Antioxidant rich Morus alba Leaf extract induces apoptosis in human colon and breast cancer cells by the downregulation of nitric oxide produced by inducible nitric oxide synthase. Nutr Cancer. 2013;65(2), 305-310.
Samrit T, Osotprasit S, Chaiwichien A, Suksomboon P, Chansap S, Athipornchai A, Changklungmoa N, Kueakha P. Cold-Pressed Sacha Inchi Oil: High in Omega-3 and Prevents Fat Accumulation in the Liver. Pharmaceuticals. 2024; 17, 220.
Puupponen PR, Nohynek L, Ammann S, Oksman CKM, Buchert J. Enzyme-assisted processing increases antimicrobial and antioxidant activity of bilberry. J. Agr. Food Chem. 2008;56, 681–688.
Cobs RM., Concha OJ, Weinstein OC, Zuniga HME. Assessment of antiproliferative activity of pectic substances obtained by different extraction methods from rapeseed cake on cancer cell lines. Carbohyd. Polym. 2015;117,923–932.
Jung SH, Han JH, Park HS, Lee DH, Kim SJ, Cho HS, Kang JS, Myung CS. Effects of unaltered and bioconverted mulberry leaf extracts on cellular glucose uptake and antidiabetic action in animals. BMC Complement. Altern. Med. 2019;19, 55.
Chan CY, Wei L, Castro MF, Koo WL. (-)-Epigallocatechin-3-gallate blocks 3T3-L1 adipose conversion by inhibition of cell proliferation and suppression of adipose phenotype expression. Life Sci. 2011;89, 779–785.
Kim M, Choi SY, Lee P, Hur JY. Neochlorogenic acid inhibits lipopolysaccharide-induced activation and pro-inflammatory responses in BV2 microglial cells. Neurochem. Res. 2015;40, 1792–1798.
Li J, Dou L, Chen SF, Zhou HH, Mou FZ. Neochlorogenic acid: An anti-HIV active compound identified by screening of Cortex Mori [Morus alba L (Moraceae)]. Pharm Biol. 2021; 59, 1517–1527.
Yu MH, Hung TW, Wang CC, Wu AW, Yang TW, Yang CY, Tseng TH, Wang JC. Neochlorogenic acid attenuates hepatic lipid accumulation and inflammation via regulating miR-34a in vitro. Int. J. Mol. Sci. 2021; 22, 13163.
Nakatani N, Kyano, S, Kikuzaki H, Sumino K, Katgairi K, Mitani T.. Identification, quantitative determination, and antioxidative activities of chlorogenic acid isomers in prune (Prunus domestica L). J. Agric. Food Chem. 2000;48, 5512–5516
Han JH, Jang KW, Myung CS. Garcinia cambogia attenuates adipogenesis by affecting CEBPB and SQSTM1/p62-mediated selective autophagic degradation of KLF3 through RPS6KA1 and STAT3 suppression. Autophagy 2022;18, 518–539.
Kandil AS, Abou EF, Shemy EHA. Cytotoxic profile activities of ethanolic and methanolic extracts of chicory plant (Cichorium intybus L.). J. Radiat. Res. Appl. Sci. 2019;12(1), 106-111.
Khan MA, Rahman AA, Islam S, Khandokhar P, Parvin S, Islam MB, Hossain M, Rashid M, Sadik G, Nasrin S. A comparative study on the antioxidant activity of methanolic extracts from different parts of Morus alba L. (Moraceae). BMC Research Notes,2013; 6(24), 24-33.
Celep E, Aydın A, Yesilada EA comparative study on the in vitro antioxidant potentials of three edible fruits: cornelian cherry, Japanese persimmon and cherry laurel. Food Chem. Toxicol. 2012;50(9):3329-3335.
Do QD, Angkawijaya AE, Tran-Nguyen PL, Huynh LH, Soetaredjo FE, Ismadji S, Ju YH. Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. J. Food Drug Anal. 2014;22(3):296-302.
Yeşiloğlu Y, Şit L. Antioxidant properties on various solvent extracts from purple basil. Spectrochim. Acta. A Mol. Biomol. Spectrosc. 2012;95:100-106.
Agustini K, Sangande F, Nuralih N, Harahap AM, Ningsih, S, Bahtiar, A. Molecular mechanism elucidation of Ocimum basilicum as anticancer using system bioinformatic approach supported by in vitro assay. Pharmacia. 2024;71, 1–12.
Al-Hadid JK, Abu IB, Harb AA., Sharab A, Alhadid A. Acacia cyanophylla, Eucalyptus camaldulensis, and Pistacia atlantica Ethanol Extracts Revealed Cytotoxicity of Breast Cancer Cell Lines: Trop. J. Nat. Prod. Res. 2021;5(10), 1808–1815.
Barbaza MYU, De Castr, CKA, Ramos JLT, Hsieh CL, Tsai PW. Synergistic Antidiabetic Activity of Extracts of Asystasia gangetica and Morus alba. Trop. J. Nat. Prod. Res. 2021; 5(2), 243–247
Muslimin L, Zainal TH, Hardianti B, Megawati M, Marwati M. Effect of Solvent Extraction on Antityrosinase and Sun Protection Factor of Mulberry (Morus alba L.) Cultivated in Wajo, Indonesia. Trop. J. Nat. Prod. Res. 2023; 7(6), 3114–3118.