The Comparison of Antioxidative and Cytotoxic Activities of Fresh and Dried Piper betle L. leave Extracts on MCF-7, HELA and SK-LU-1

Phuoc Vo Thi

University of Sciences, Hue University, Vietnam.

Tam Nguyen Thi

University of Sciences, Hue University, Vietnam.

Minh Tri Nguyen

University of Sciences, Hue University, Vietnam.

Dieu Ngan Phan Thi

University of Sciences, Hue University, Vietnam.

Nha Khue Than Trong

University of Sciences, Hue University, Vietnam.

Cam Ha Che Thi *

University of Sciences, Hue University, Vietnam.

*Author to whom correspondence should be addressed.


Abstract

The use of herbal medicines produced from plants and herbs, as one element of complementary and alternative medicine, is still increasing worldwide. In this study, Piper betle L. leaf in two states, fresh and dried, was tested to explore its compounds and total phenolic and flavonoid contents. From there evaluated its antioxidant, and cytotoxic activities In vitro. Gas chromatography–mass spectrometry is a method for identifying compounds (GC-MS). The total phenolic content was measured by using Folin-Ciocalteu method and the total flavonoid content was based on a complex formation with AlCl3. The antioxidant activity was evaluated by DPPH. The cytotoxic activity was performed against MCF-7, Hela, and SK-LU-1 cancer cell lines using sulforhodamine B assay. Toxicity testing for normal cells was also investigated. Results show the dried betel leaf extract (DBLE) contains eugenol (37.56%) as the primary compound, while 2.5-dimethylbenzoic acid (89.58%) was the primary compound in fresh betel leaf extract (FBLE). The extracts are rich polyphenols and flavonoids and have strong antioxidant activity. The polyphenol content in DBLE (240.9 mg GAE/g) is higher than FBLE (165.2 mg GAE/g). Especially, the DBLE has shown significant DPPH scavenging activity (IC50 = 3.21 µg/mL) when compared with standard ascorbic acid (IC50 = 6.71 µg/mL) and FBLE (IC50 = 22.97 µg/mL). The cytotoxic activity of FBLE against MCF-7 and Hela cell lines shows IC50 value above 50 µg/mL, while DBLE shows IC50 value of 21.88 µg/mL and 26.68 µg/mL, respectively, with no difference for the SK-LU-1 line (IC50 values from 35.0 to 38.0 μg/mL). There is a positive correlation between the phenolic content, antioxidant, and cytotoxic activities of the extracts. In conclusion, DBLE exhibits higher antioxidant and cytotoxic activities than FBLE.

Keywords: Betel leaf (Piper betle), fresh and dried leaves, antioxidant, cytotoxicity


How to Cite

Thi, Phuoc Vo, Tam Nguyen Thi, Minh Tri Nguyen, Dieu Ngan Phan Thi, Nha Khue Than Trong, and Cam Ha Che Thi. 2023. “The Comparison of Antioxidative and Cytotoxic Activities of Fresh and Dried Piper Betle L. Leave Extracts on MCF-7, HELA and SK-LU-1”. Asian Journal of Research in Botany 6 (2):339-51. https://journalajrib.com/index.php/AJRIB/article/view/201.

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References

Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians. 2021; 71(3):209- 249.

Hashem S, Ali TA, Akhtar S, Nisar S, Sageena G, Ali S, Elfaki I. Targeting cancer signaling pathways by natural products: Exploring promising anti-cancer agents. Biomedicine Pharmacotherapy. 2022;150:113054.

Rintu D, Shinjini M, Kaustab M, Pramathadhip P, Umesh P, Banerjee E. Anti- oxidant and anti-inflammatory activities of different varieties of Piper leaf extracts (Piper betle L.). Nutrition and Food Sciences. 2015;5:415.

Majumdar AG, Subramanian M. Hydroxychavicol from Piper betle induces apoptosis, cell cycle arrest, and inhibits epithelial-mesenchymal transition in pancreatic cancer cells. Biochemical pharmacology. 2019;166:274-291.

Wu PF, Tseng HC, Chyau CC, Chen JH, Chou FP. Piper betle leaf extracts induced human hepatocellular carcinoma Hep3B cell death via MAPKs regulating the p73 pathway in vitro and in vivo. Food function. 2014;5(12):3320-3328.

Sanubol A, Chaveerach A, Tanee T, Sudmoon R. Pre-clinical evaluation of extracts and essential oils from betel-like scent Piper species identified potential cancer treatment. Afr J Tradit Complement Altern Med. 2017;14(1):89-102.

Das S, Ray A, Nasim N, Nayak S, Mohanty S. Effect of different extraction techniques on total phenolic and flavonoid contents, and antioxidant activity of betelvine and quantification of its phenolic constituents by validated HPTLC method. 3 Biotech. 2019;9(1):1-8.

Abrahim NN, Kanthimathi M, Abdul-Aziz AJB. Piper betle shows antioxidant activities, inhibits MCF-7 cell proliferation and increases activities of catalase and superoxide dismutase. BMC Complementary Alternative Medicine. 2012;12(1):1-11.

Thị CHC, Nguyễn HĐ, Lê Hoàng DMJH. Influence of Piper betle L. extract on umbilical cord cells in vitro and potential treating cutaneous wound. Heliyon. 2021; 7(3).

Valle DL, Puzon JJM, Cabrera EC, Rivera WL. Thin Layer Chromatography-Bioautography and Gas Chromatography-Mass Spectrometry of Antimicrobial Leaf Extracts from Philippine Piper betle L. against Multidrug- Resistant Bacteria. Evidence-Based Complementary and Alternative Medicine. 2016;4976791.

Murugesan K, Ilanchezhian T, Vijayan H. The Effect of Hydroethanolic Solvents Extracts of Piper betel and Plectranthus amboinicus on Food Poison Disease Causing Pathogenic Bacteria. International Journal of Pharmaceutical Sciences Review and Research. 2017;45(1):257-261.

Pratiwi NPRK, Muderawan IW. Analisis kandungan kimia ekstrak daun sirih hijau (Piper betle) dengan GC-MS. Prosiding Seminar Nasional MIPA; 2016.

Selvaraj GK, Wilson JJ, Kanagaraj N, Subashini E, Thangavel S. Enhanced antifungal activity of Piper betle against candidiasis infection causing Candida Albicans and In silico analysis with its virulent protein. Biomedical and Biotechnology Research Journal. 2022; 6(1):73-80.

Jenie B, Apryiantono A. Antibacterial activity of green sirih (Piper betle L) extract towards food pathogens. Jurnal Teknologi dan Industri Pangan. 2008;19(1);1-1.

Annegowda H, Tan P, Mordi M, Ramanathan S, Hamdan M, Sulaiman M, Mansor S. TLC–bioautography-guided isolation, HPTLC and GC–MS-assisted analysis of bioactives of Piper betle leaf extract obtained from various extraction techniques: In vitro evaluation of phenolic content, antioxidant and antimicrobial activities. Food Analytical Methods. 2013; 6(3):715-726.

Das S, Parida R, Sandeep IS, Kar B, Nayak S, Mohanty S. Chemical composition and antioxidant activity of some important betel vine landraces. Biologia. 2016;71(2):128-132.

Nguyen LTT, Nguyen TT, Nguyen HN, Bui QTP. Simultaneous determination of active compounds in Piper betle Linn. leaf extract and effect of extracting solvents on bioactivity. Engineering Reports. 2020; 2(10):e12246.

Dương HT, Loan NH, Ngân PTK, Thư LHA, Dũng PT, Thành NVK, Kiều NKT. Evaluation of biological activity of betel leaf extract (Piper betle L.) obtained by ultrasonic extraction method. Vietnam Science and Technology Magazine. 2022; 64(3):37-42.

Harini SS, Sougandhi PR, Tenkayala DSR, Gopinath KR. Antioxidant activity (Phenol and flavonoid content) of three different cultivars of Piper Betle L. (Piperaceae). Journal of Drug Delivery Therapeutics. 2018;8(5-s):143-148.

Boontha S, Taowkaen J, Phakwan T, Worauaicha T, Kamonnate P, Buranrat B, Pitaksuteepong T. Evaluation of antioxidant and anticancer effects of Piper betle L (Piperaceae) leaf extract on MCF-7 cells, and preparation of transdermal patches of the extract. Tropical Journal of Pharmaceutical Research. 2019;18(6): 1265-1272.

Sartini S, Khaerawati N, Kamril RA, Febriani NJP. The effects of fresh leaf-to- water ratio and heating time on the antifungal and antioxidant activities of betel leaf (Piper betle L.) extract. 2020;10(1): 117-124.

Alam MB, Park NH, Song BR, Lee SHJA. Antioxidant Potential-Rich Betel Leaves (Piper betle L.) Exert Depigmenting Action by Triggering Autophagy and Downregulating MITF/Tyrosinase In Vitro and In vivo. 2023;12(2):374.

Fathilah A, Sujata R, Norhanom A, Adenan M. Antiproliferative activity of aqueous extract of Piper betle L. and Psidium guajava L. on KB and HeLa cell lines. Journal of Medicinal Plants Research. 2010;4(11):987-990.

Fathy M, Fawzy MA, Hintzsche H, Nikaido T, Dandekar T, Othman EM. Eugenol exerts apoptotic effect and modulates the sensitivity of HeLa cells to cisplatin and radiation. Molecules. 2019;24(21):3979.

De Sa Junior PL, Câmara DAD, Costa AS, Ruiz JLM, Levy D, Azevedo RA, Pessoa NDS. Apoptotic effect of eugenol envolves G2/M phase abrogation accompanied by mitochondrial damage and clastogenic effect on cancer cell in vitro. Phytomedicine. 2016;23(7):725-735.

Liang WZ, Chou CT, Hsu SS, Liao WC, Shieh P, Kuo DH, Jan CR. The involvement of mitochondrial apoptotic pathway in eugenol-induced cell death in human glioblastoma cells. Toxicology letters. 2015;232(1):122-132.

Zari AT, Zari TA, Hakeem KR. Anticancer Properties of Eugenol: A Review. Molecules, 2021;26(23):7407.

Aara A, Chappidi V, Ramadas MN, Care P. Antioxidant activity of eugenol in Piper betel leaf extract. Journal of Family Medicine and Primary Care. 2020;9(1): 327.

Gushiken LFS, Beserra FP, Hussni MF, Gonzaga MT, Ribeiro VP, De Souza PF, Takahira RK. Beta-caryophyllene as an antioxidant, anti-inflammatory and re-epithelialization activities in a rat skin wound excision model. Oxidative medicine cellular longevity. 2022;9004014.

Vanin AB, Orlando T, Piazza SP, Puton B, Cansian RL, Oliveira D, Paroul N. Antimicrobial and antioxidant activities of clove essential oil and eugenyl acetate produced by enzymatic esterification. Applied Biochemistry Biotechnology. 2014; 174(4):1286-1298.

Irrera N, D’Ascola A, Pallio G, Bitto A, Mannino F, Arcoraci V, Metro D. β- caryophyllene inhibits cell proliferation through a direct modulation of CB2 receptors in glioblastoma cells. Cancers. 2020;12(4):1038.

Lei J, Wang Q, Li G, Li Y, Zhang P, Xu G. β-Caryophyllene from Chilli Pepper Inhibits the Proliferation of Non-Small Cell Lung Cancer Cells by Affecting miR-659-3p- Targeted Sphingosine Kinase 1 (SphK1). International Journal of General Medicine. 2021;14:9599.

Chen H, Yuan J, Hao J, Wen Y, Lv Y, Chen L, Yang X. α-Humulene inhibits hepatocellular carcinoma cell proliferation and induces apoptosis through the inhibition of Akt signaling. Food Chemical Toxicology. 2019;134:110830.

Legault J, Pichette A. Potentiating effect of β‐caryophyllene on anticancer activity of α‐humulene, Isocaryophyllene and paclitaxel. Journal of Pharmacy Pharmacology. 2007;59(12):1643-1647.

Losada-Echeberría M, Herranz-López M, Micol V, Barrajón-Catalán E. Polyphenols as promising drugs against main breast cancer signatures. Antioxidants. 2017;6(4): 88.