Ethyl acetate crude fraction of Macaranga subpeltata and silymarin increase Vero cell survival and HepG2 cell death due to oxidative stress

Harto Widodo; Abdul Rohman

doi:10.7324/JAPS.2023.84845

Abstract

Reactive oxygen species (ROS) trigger oxidative stress (OS) that determines the onset of aging and various disease progressions including cancer. Macaranga subpeltata is among medicinal plants, which shows an important source of antioxidant (AO) compounds and has the potency to overcome those problems. Silymarin is a plant-derived flavonolignan with tremendous health benefits as an AO and organ protector. Hydrogen peroxide (H2O2), an active prooxidant molecule, enables the generation of OS in mammalian cells. This study aims to evaluate the effect of ethyl acetate crude fraction of M. subpeltata (EACFM) and silymarin in H2O2-induced OS of Vero and HepG2 cell lines. The 3-(4,5-dimethyl azole-2-yl)-2,5-diphenyltetrazolium bromide assay was used to evaluate cell viability, while examination of ROS generation and cell death were done using the flow cytometric method. The study revealed that H2O2 induced ROS generation in a dose-dependent manner both on Vero and HepG2 cell lines, and the HepG2 hepatocarcinoma cells were more resistant to H2O2 toxicity than the Vero cells. Silymarin suppressed ROS formation on both H2O2-treated cell lines with different effects; it alleviated H2O2 intoxication on Vero cells but enhanced the toxicity of H2O2 on HepG2. The EACFM reduced H2O2 intoxications of Vero cells and modulated ROS-mediated cell death of HepG2 cells through either apoptosis or necrosis. The EACFM showed a higher capacity to reduce OS due to H2O2 treatment on Vero cells; however, silymarin showed better anticancer properties.

Keyword: Antioxidant apoptosis anticancer cell death hydrogen peroxide necrosis

Citation:

Widodo H, Rohman A. Ethyl acetate crude fraction of Macaranga subpeltata and silymarin increase Vero cell survival and HepG2 cell death due to oxidative stress. J Appl Pharm Sci, 2023. https://doi.org/10.7324/JAPS.2023.84845

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reference

CCRC. Cancer Chemoprevention Reserch Centre working manual. Fakultas Farmasi UGM, Yogyakarta, Indonesia, 2019.

Cheong CU, Yeh CS, Hsieh YW, Lee YR, Lin MY, Chen CY, Lee CH. Protective effects of costunolide against hydrogen peroxide-induced injury in PC12 cells. Molecules, 2016; 21(898):1-0. https://doi.org/10.3390/molecules21070898

Chidawanyika T, Supattapone S. Hydrogen peroxide-induced cell death in mammalian cells. J Cell Signal, 2021; 2(3):206-11. https://doi.org/10.33696/Signaling.2.052

Eruslanov E, Kusmartsev S. Identification of ROS using oxidized DCFDA and flow-cytometry. In: Armstrong D. (Ed.). Advanced protocols in oxidative stress II. Methods in molecular biology (methods and protocols), Humana Press, Totowa, NJ, p 594, 2010. https://doi.org/10.1007/978-1-60761-411-1_4

Fallah M, Davoodvandi A, Nikmanzar S, Aghili S, Mirazimi SMA, Aschner M, Rashidian A, Hamblin MR, Chamanara M, Naghsh N, Mirzaei H. Silymarin (milk thistle extract) as a therapeutic agent in gastrointestinal cancer. Biomed Pharmacother, 2021; 142:112024. https://doi.org/10.1016/j.biopha.2021.112024

Habtemariam S, Lentini G. Plant-derived anticancer agents: Lesson from the pharmacology of geniposide and its aglycon, genipin. Biomedicines, 2018; 6(2):39. https://doi.org/10.3390/biomedicines6020039

Haryanti S, Rahmawati N, Sholikhah IYM, Widiyastuti Y. Ficus septica, an ecosystem keystone species induced ROS mediated cytotoxicity in HepG2 hepatocarcinoma cells. The 8th International Conference on Sustainable Agriculture and Environment. IOP Conf Ser Earth Environ Sci, 2021; 905:012101. https://doi.org/10.1088/1755-1315/905/1/012101

Jiang HH, Yan FS, Shen L, Ji HF. Silymarin versus silibinin: differential antioxidant and neuroprotective effects against H2O2-induced oxidative stress in PC12 cells. Nat Prod Commun, 2016; 11(5):633-6. https://doi.org/10.1177/1934578X1601100520

Khazaei R, Seidavi A, Bouyeh M. A review on the mechanisms of the effect of silymarin in milk thistle (Silybum marianum) on some laboratory animals. Vet Med Sci, 2022; 8;289-301. https://doi.org/10.1002/vms3.641

Kim H, Lee GR, Kim J, Baek JY, Jo YJ, Hong SE, Kim SH, Lee J, Lee HI, Park SK, Kim HM, Lee HJ, Chang TS, Rhee SG, Lee JS, Jeong W. Sulfiredoxin inhibitor induces preferential death of cancer cells through reactive oxygen species-mediated mitochondrial damage. Free Radic Biol Med, 2016; 91:264-74. https://doi.org/10.1016/j.freeradbiomed.2015.12.023

Liau LL, Suzana M, Azurah AGN, Chua KH. Hydrogen peroxide induces acute injury and up-regulates inflammatory gene expression in hepatocytes: an in vitro model. Sains Malays, 2016; 45(3):451-8.

Mili? N, Miloševi? N, Suvajdži? L, Žarkov M, Abenavoli L. New therapeutic potentials of milk thistle (Silybum marianum). Nat Prod Commun, 2013; 8(12):1801-10. https://doi.org/10.1177/1934578X1300801236

Nakai K, Tsuruta D. What are reactive oxygen species, free radicals, and oxidative stress in skin diseases? Int J Mol Sci, 2021; 22:10799. https://doi.org/10.3390/ijms221910799

Nakamura H, Takada K. Reactive oxygen species in cancer: current findings and future directions. Cancer Sci, 2021; 112:3945-52. https://doi.org/10.1111/cas.15068

Ndombera FT. Anti-cancer agents and reactive oxygen species modulators that target cancer cell metabolism. Pure Appl Chem, 2017; 89(9):1333-48. https://doi.org/10.1515/pac-2016-1219

Perillo B, Donato MD, Pezone A, Zazzo ED, Giovannelli P, Galasso G, Castoria G, Migliaccio A. ROS in cancer therapy: the bright side of the moon. Exp Mol Med, 2020; 52:192-203. https://doi.org/10.1038/s12276-020-0384-2

Rhazi H, Safini N, Mikou K, Alhyane M, Tadlaoui KO, Lin X, Venkatesan NP, Elharrak M. Production of small ruminant morbillivirus, rift valley fever virus and lumpy skin disease virus in CelCradle™-500A bioreactors. BMC Vet Res, 2021; 17:93. https://doi.org/10.1186/s12917-021-02801-4

Sefried S, Ha¨ring HU, Weigert C, Eckstein SS. Suitability of hepatocyte cell lines HepG2, AML12 and THLE-2 for investigation of insulin signaling and hepatokine gene expression. Open Biol, 2018; 8:180147. https://doi.org/10.1098/rsob.180147

Sène MA, Xia Y, Kamen AA. Overview of recent advances in Vero cells genomic characterization and engineering for high-throughput vaccine manufacturing. Clin Transl Disc, 2022; 2:e40. https://doi.org/10.1002/ctd2.40

Srinivasan V, Panneerselvam R, Gunasekaran S, Palani S. Nephro-protective activity of ethanolic extract of Melia azadirachta against H2O2 induced toxicity in Vero cell line. IJABPT, 2015; 6(4):44-9.

Sudrajad H, Widodo H, Fauzi, Widiyastuti Y. Optimizing cultivation technique of Silybum marianum (L.) Gaertn. under tropical climate. Indian J Agric Res, 2022; 56(1):76-80. https://doi.org/10.18805/IJARe.A-665

Surai PF. Silymarin as a natural antioxidant: an overview of the current evidence and perspectives. Antioxidant, 2015; 4:204-47. https://doi.org/10.3390/antiox4010204

U?ran R, Levent A. Chapter 3: oxidative stress and liver. In: Güven A (Ed.). Oxidative stress and antioxidant defense system, Livre de Lyon, Lyon, France, vol. 1, p 92, 2021.

Vermes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptosis cell death. J Immunol Methods, 2000; 243(1-2):169-90. https://doi.org/10.1016/S0022-1759(00)00233-7

Widodo H, Sismindari S, Asmara W, Rohman A. Antioxidant activities of methanolic extract and its fractions of Baccaurea racemosa and Macaranga subpeltata leaves. Food Res, 2020; 4(1):127-34. https://doi.org/10.26656/fr.2017.4(1).144

Widodo H, Sismindari S, Asmara W, Rohman A. Antioxidant activity, total phenolic and flavonoid contents of selected medicinal plants used for liver diseases and its classification with chemometrics. J Appl Pharm Sci, 2019; 9(06):099-105. https://doi.org/10.7324/JAPS.2019.90614

Wu JP, Tsai CC, Yu-Lan Yeh YL, Lin YM, Lin CC, Day CH, Shen CY, Padma VV, Pan LF, Huang CY. Silymarin accelerates liver regeneration after partial hepatectomy. Evid Based Complement Altern Med, 2015; 603529:14. https://doi.org/10.1155/2015/603529

Yan D, Cui H, Zhu W, Talbot A, Zhang LG, Sherman JH, Keidar M. The strong cell-based hydrogen peroxide generation triggered by cold atmospheric plasma. Sci Rep, 2017; 7(10831):1-9. https://doi.org/10.1038/s41598-017-11480-x

Yang H, Villani RM, Wang H, Simpson MJ, Roberts MS, Tang M, Liang X. The role of cellular reactive oxygen species in cancer chemotherapy. J Exp Clin Cancer Res, 2018; 37:266. https://doi.org/10.1186/s13046-018-0909-x

Zenin V, Ivanova J, Pugovkina N, Shatrova A, Aksenov N, Tyuryaeva I, Kirpichnikova K, Kuneev I, Zhuravlev A, Osyaeva E, Lyublinskaya E, Gazizova I, Guriev N, Olga Lyublinskaya O. Resistance to H2O2-induced oxidative stress in human cells of different phenotypes. Redox Biol, 2022; 50:102245. https://doi.org/10.1016/j.redox.2022.102245

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