Antioxidant and Cytotoxic Effects of Aqueous Extract of Iranian Green Tea (Camellia sinensis L.) Antioxidant and Cytotoxicity of Camellia sinensis
Iranian Journal of Pharmaceutical Sciences,
Vol. 18 No. 1 (2022),
15 Dey 2022
,
Page 77-86
https://doi.org/10.22037/ijps.v18.41381
Abstract
Green tea, a common beverage, has several pharmacological effects. In this study, the amounts of phenolic compounds and epigallocatechin (EGC) were determined in an aqueous extract of green tea planted in Lahijan (a city in the north of Iran) by Folin-Ciocalteu method and HPLC, respectively. Antioxidant activity of the extract was measured by DPPH and the FRAP assays. Furthermore, its cytotoxic effect was investigated on HT-29 (colorectal cancer) and 3T3 (normal fibroblast) cell lines, after 24 and 48 h treatment by MTT method. Trypan blue dye exclusion measurement was also done on the HT-29 cells treated with the extract. The results have shown that each gram of the dried extract contains 283.6 ± 10.57 mg total phenolic content and 88.44 ± 1.85 mg EGC. The antioxidant activity of the extract measured by DPPH showed an EC50 value of 84.31 ± 4.14 μg/ml. In the FRAP test, the equivalent amount of Iron (Fe) was 4.45 mmol per g of the dried extract. The extract showed toxicity on the HT-29 cell line with an IC50 value of 82.45 ± 18.39 μg/ml after 48 h treatment. The data were confirmed by trypan blue dye exclusion test. Based on the results, adding green tea to the diet may have many health benefits.
Keywords:
- Antioxidant,
- Antiproliferative,
- Catechin,
- DPPH,
- EGC,
- FRAP,
- Green tea.
How to Cite
Mitra Mehrabani, Bagher Amirheidari, Negar Ahmadinia, Masoud Forooghi-Nematollahi, & Salehe Sabouri. (2022). Antioxidant and Cytotoxic Effects of Aqueous Extract of Iranian Green Tea (Camellia sinensis L.): Antioxidant and Cytotoxicity of Camellia sinensis. Iranian Journal of Pharmaceutical Sciences, 18(1), 77–86. https://doi.org/10.22037/ijps.v18.41381
References
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[18] Zhao LG, Li HL, Sun JW, Yang Y, Ma X, Shu XO, Zheng W, Xiang YB. Green tea consumption and cause-specific mortality: Results from two prospective cohort studies in China. J. Epidemiol. (2017) 27: 36-41.
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[21] Azadi Gonbad R, Afzan A, Karimi E, Sinniah UR, Kumara Swamy M. Phytoconstituents and antioxidant properties among commercial tea (Camellia sinensis L.) clones of Iran. Electron. J. Biotechnol. (2015) 18: 433-8.
[22] Saboura A, Ahmadi A, Zeynali A, Parsa M. Comparison between the contents of phenolic and flavonoid compounds and aerial part antioxidant activity in Scutellaria pinnatifida in two northiranian populations. J. Rafsanjan Univ. Med. Sci. (2014) 13: 249-66.
[23] Moraes-de-Souza R, Oldoni T, Regitano-d'Arce M , Alencar S. Antioxidant activity and phenolic composition of herbal infusions consumed in Brazil. CYTA-J. Food (2008) 6: 41-7.
[24] Sharma V, Gulati A, Ravindranath SD, Kumar V. A simple and convenient method for analysis of tea biochemicals by reverse phase HPLC. J. Food Compos. Anal. (2005) 18: 583-94.
[25] Khurm M, Chaudhry BA, Uzair M , Janbaz KH. Antimicrobial, Cytotoxic, Phytotoxic and Antioxidant Potential of Heliotropium strigosum Willd. Medicines (2016) 3: 20.
[26] Tsao R, Yang R, Young JC. Antioxidant isoflavones in osage orange, Maclura pomifera (Raf.) Schneid. J. Agric. Food Chem. (2003) 51: 6445-51.
[27] Abaszadeh M, Ebrahimi A , Sabouri S. Investigating the in vitro antiproliferative and apoptosis-inducing effects of pyranochromene derivatives. Biointerface Res. Appl. Chem. (2021) 11: 10987-95.
[28] Strober W. Trypan blue exclusion test of cell viability. Curr. Protoc. Immunol. (2015) 111: A3.B.1-A3.B.3.
[29] Komes D, Horžić D, Belščak A, Ganić KK, Vulić I. Green tea preparation and its influence on the content of bioactive compounds. Food Res. Int. (2010) 43: 167-76.
[30] Rababah TM, Hettiarachchy NS , Horax R. Total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone. J. Agric. Food Chem. (2004) 52: 5183-6.
[31] Satoh E, Tohyama N, Nishimura M. Comparison of the antioxidant activity of roasted tea with green, oolong, and black teas. Int. J. Food Sci. Nutr. (2005) 56: 551-9.
[32] Toschi TG, Bordoni A, Hrelia S, Bendini A, Lercker G , Biagi PL. The protective role of different green tea extracts after oxidative damage is related to their catechin composition. J. Agric. Food Chem. (2000) 48: 3973-8.
[33] Zuo Y, Chen H, Deng Y. Simultaneous determination of catechins, caffeine and gallic acids in green, Oolong, black and pu-erh teas using HPLC with a photodiode array detector. Talanta (2002) 57: 307-16.
[34] Oh J, Jo H, Cho AR, Kim S-J, Han J. Antioxidant and antimicrobial activities of various leafy herbal teas. Food Cont. (2013) 31: 403-9.
[35] Quan PT, Hang T, Ha NH, Glang B. Total polyphenols, total catechin content and DPPH free radical scavenger activity of several types of Vietnam commercial green tea. Sci. Technol. Dev. (2007) 10: 5-11.
[36] Farooq S, Sehgal A. Synergistic antioxidant interactions between green tea and Ocimum gratissimum. Asian. Pac. J. Trop. Biomed. (2019) 9: 333-8.
[37] Benzie IF, Szeto Y. Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay. J. Agric. Food Chem. (1999) 47: 633-6.
[38] Wang L, Wei K, Jiang Y, Cheng H, Zhou J, He W , Zhang C. Seasonal climate effects on flavanols and purine alkaloids of tea (Camellia sinensis L.). Eur. Food Res. Technol. (2011) 233: 1049-55.
[39] Wei K, Wang L, Zhou J, He W, Zeng J, Jiang Y , Cheng H. Catechin contents in tea (Camellia sinensis) as affected by cultivar and environment and their relation to chlorophyll contents. Food Chem. (2011) 125: 44-8.
[40] Xu Y-Q, Ji W-B, Yu P, Chen J-X, Wang F, Yin J-F. Effect of extraction methods on the chemical components and taste quality of green tea extract. Food Chem. (2018) 248: 146-54.
[41] Ramirez-Mares MV, Chandra S, de Mejia EG. In vitro chemopreventive activity of Camellia sinensis, Ilex paraguariensis and Ardisia compressa tea extracts and selected polyphenols. Mutat. Res. (2004) 554: 53-65.
[42] Shah S, Gani A, Ahmad M, Shah A, Gani A , Masoodi F. In vitro antioxidant and antiproliferative activity of microwave-extracted green tea and black tea (Camellia sinensis): a comparative study. NutraFoods (2015) 14: 207-15.
[43] Zhao X, Song JL, Kim JD, Lee JS, Park KY. Fermented Pu-erh tea increases in vitro anticancer activities in HT-29 cells and has antiangiogenetic effects on HUVECs. J. Environ. Pathol. Toxicol. Oncol. (2013) 32: 275-88.
[44] Lambert JD, Lee MJ, Diamond L, Ju J, Hong J, Bose M, Newmark HL, Yang CS. Dose-dependent levels of epigallocatechin-3-gallate in human colon cancer cells and mouse plasma and tissues. Drug Metab. Dispos. (2006) 34: 8-11.
[45] Valcic S, Timmermann BN, Alberts DS, Wächter GA, Krutzsch M, Wymer J, Guillén JM. Inhibitory effect of six green tea catechins and caffeine on the growth of four selected human tumor cell lines. Anticancer Drugs (1996) 7: 461-8.
[46] Ravindranath MH, Saravanan TS, Monteclaro CC, Presser N, Ye X, Selvan SR , Brosman S. Epicatechins purified from green tea (Camellia sinensis) differentially suppress growth of gender-dependent human cancer cell lines. Evid. Based Complement. Altern. Med. (2006) 3: 237-47.
[2] Hajiaghaalipour F, Kanthimathi MS, Sanusi J , Rajarajeswaran J. White tea (Camellia sinensis) inhibits proliferation of the colon cancer cell line, HT-29, activates caspases and protects DNA of normal cells against oxidative damage. Food Chem. (2015) 169: 401-10.
[3] Cabrera C, Artacho R , Gimenez R. Beneficial effects of green tea_a review. J. Am. Coll. Nutr. (2006) 25: 79-99.
[4] Pastore RL , Fratellone P. Potential health benefits of green tea (Camellia sinensis): a narrative review. Explore (2006) 2: 531-9.
[5] Chung MS, Bae WJ, Choi SW, Lee KW, Jeong HC, Bashraheel F, Jeon SH, Jung JW, Yoon BI, Kwon EB, Oh HA, Hwang SY, Kim SW. An Asian traditional herbal complex containing Houttuynia cordata Thunb, Perilla frutescens Var. acuta and green tea stimulates hair growth in mice. BMC Complement. Altern. Med. (2017) 17: 515-25.
[6] Chopade V, Phatak A, Upaganlawar A , Tankar A. Green tea (Camellia sinensis): Chemistry, traditional, medicinal uses and its pharmacological activities-a review. Pharmacogn. Rev. (2008) 2: 157-62.
[7] Xu J, Xu Z , Zheng W. A Review of the Antiviral Role of Green Tea Catechins. Molecules (2017) 22: 1337.
[8] Miyata Y, Matsuo T, Araki K, Nakamura Y, Sagara Y, Ohba K , Sakai H. Anticancer effects of green tea and the underlying molecular mechanisms in bladder cancer. Medicines (2018) 5: 87.
[9] Chong SY, Chiang HY, Chen TH, Liang YJ , Lo YC. Green tea extract promotes DNA repair in a yeast model. Sci. Rep. (2019) 9: 3842.
[10] Yu GP, Hsieh CC, Wang LY, Yu SZ, Li XL , Jin TH. Green-tea consumption and risk of stomach cancer: a population-based case-control study in Shanghai, China. Cancer Causes Control (1995) 6: 532-8.
[11] Yuan JM, Koh WP, Sun CL, Lee HP, Yu MC. Green tea intake, ACE gene polymorphism and breast cancer risk among Chinese women in Singapore. Carcinogenesis (2005) 26: 1389-94.
[12] Nagle CM, Olsen CM, Bain CJ, Whiteman DC, Green AC, Webb PM. Tea consumption and risk of ovarian cancer. Cancer Causes Control (2010) 21: 1485-91.
[13] Liu J, Liu S, Zhou H, Hanson T, Yang L, Chen Z , Zhou M. Association of green tea consumption with mortality from all-cause, cardiovascular disease and cancer in a Chinese cohort of 165,000 adult men. Eur. J. Epidemiol. (2016) 31: 853-65.
[14] Fujiki H, Sueoka E, Watanabe T , Suganuma M. Primary cancer prevention by green tea, and tertiary cancer prevention by the combination of green tea catechins and anticancer compounds. J. Cancer Prev. (2015) 20: 1-4.
[15] Guo Y, Zhi F, Chen P, Zhao K, Xiang H, Mao Q, Wang X, Zhang X. Green tea and the risk of prostate cancer: A systematic review and meta-analysis. Medicine (2017) 96: e6426.
[16] Borrelli F, Capasso R, Russo A, Ernst E. Systematic review: green tea and gastrointestinal cancer risk. Aliment. Pharmacol. Ther. (2004) 19: 497-510.
[17] Kuriyama S, Shimazu T, Ohmori K, Kikuchi N, Nakaya N, Nishino Y, Tsubono Y, Tsuji I. Green tea consumption and mortality due to cardiovascular disease, cancer, and all causes in Japan: the Ohsaki study. JAMA (2006) 296: 1255-65.
[18] Zhao LG, Li HL, Sun JW, Yang Y, Ma X, Shu XO, Zheng W, Xiang YB. Green tea consumption and cause-specific mortality: Results from two prospective cohort studies in China. J. Epidemiol. (2017) 27: 36-41.
[19] Seely D, Mills EJ, Wu P, Verma S, Guyatt GH. The effects of green tea consumption on incidence of breast cancer and recurrence of breast cancer: a systematic review and meta-analysis. Integr. Cancer Ther. (2005) 4: 144-55.
[20] Kikuchi N, Ohmori K, Shimazu T, Nakaya N, Kuriyama S, Nishino Y, Tsubono Y, Tsuji I. No association between green tea and prostate cancer risk in Japanese men: the Ohsaki Cohort Study. Br. J. Cancer (2006) 95: 371-3.
[21] Azadi Gonbad R, Afzan A, Karimi E, Sinniah UR, Kumara Swamy M. Phytoconstituents and antioxidant properties among commercial tea (Camellia sinensis L.) clones of Iran. Electron. J. Biotechnol. (2015) 18: 433-8.
[22] Saboura A, Ahmadi A, Zeynali A, Parsa M. Comparison between the contents of phenolic and flavonoid compounds and aerial part antioxidant activity in Scutellaria pinnatifida in two northiranian populations. J. Rafsanjan Univ. Med. Sci. (2014) 13: 249-66.
[23] Moraes-de-Souza R, Oldoni T, Regitano-d'Arce M , Alencar S. Antioxidant activity and phenolic composition of herbal infusions consumed in Brazil. CYTA-J. Food (2008) 6: 41-7.
[24] Sharma V, Gulati A, Ravindranath SD, Kumar V. A simple and convenient method for analysis of tea biochemicals by reverse phase HPLC. J. Food Compos. Anal. (2005) 18: 583-94.
[25] Khurm M, Chaudhry BA, Uzair M , Janbaz KH. Antimicrobial, Cytotoxic, Phytotoxic and Antioxidant Potential of Heliotropium strigosum Willd. Medicines (2016) 3: 20.
[26] Tsao R, Yang R, Young JC. Antioxidant isoflavones in osage orange, Maclura pomifera (Raf.) Schneid. J. Agric. Food Chem. (2003) 51: 6445-51.
[27] Abaszadeh M, Ebrahimi A , Sabouri S. Investigating the in vitro antiproliferative and apoptosis-inducing effects of pyranochromene derivatives. Biointerface Res. Appl. Chem. (2021) 11: 10987-95.
[28] Strober W. Trypan blue exclusion test of cell viability. Curr. Protoc. Immunol. (2015) 111: A3.B.1-A3.B.3.
[29] Komes D, Horžić D, Belščak A, Ganić KK, Vulić I. Green tea preparation and its influence on the content of bioactive compounds. Food Res. Int. (2010) 43: 167-76.
[30] Rababah TM, Hettiarachchy NS , Horax R. Total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone. J. Agric. Food Chem. (2004) 52: 5183-6.
[31] Satoh E, Tohyama N, Nishimura M. Comparison of the antioxidant activity of roasted tea with green, oolong, and black teas. Int. J. Food Sci. Nutr. (2005) 56: 551-9.
[32] Toschi TG, Bordoni A, Hrelia S, Bendini A, Lercker G , Biagi PL. The protective role of different green tea extracts after oxidative damage is related to their catechin composition. J. Agric. Food Chem. (2000) 48: 3973-8.
[33] Zuo Y, Chen H, Deng Y. Simultaneous determination of catechins, caffeine and gallic acids in green, Oolong, black and pu-erh teas using HPLC with a photodiode array detector. Talanta (2002) 57: 307-16.
[34] Oh J, Jo H, Cho AR, Kim S-J, Han J. Antioxidant and antimicrobial activities of various leafy herbal teas. Food Cont. (2013) 31: 403-9.
[35] Quan PT, Hang T, Ha NH, Glang B. Total polyphenols, total catechin content and DPPH free radical scavenger activity of several types of Vietnam commercial green tea. Sci. Technol. Dev. (2007) 10: 5-11.
[36] Farooq S, Sehgal A. Synergistic antioxidant interactions between green tea and Ocimum gratissimum. Asian. Pac. J. Trop. Biomed. (2019) 9: 333-8.
[37] Benzie IF, Szeto Y. Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay. J. Agric. Food Chem. (1999) 47: 633-6.
[38] Wang L, Wei K, Jiang Y, Cheng H, Zhou J, He W , Zhang C. Seasonal climate effects on flavanols and purine alkaloids of tea (Camellia sinensis L.). Eur. Food Res. Technol. (2011) 233: 1049-55.
[39] Wei K, Wang L, Zhou J, He W, Zeng J, Jiang Y , Cheng H. Catechin contents in tea (Camellia sinensis) as affected by cultivar and environment and their relation to chlorophyll contents. Food Chem. (2011) 125: 44-8.
[40] Xu Y-Q, Ji W-B, Yu P, Chen J-X, Wang F, Yin J-F. Effect of extraction methods on the chemical components and taste quality of green tea extract. Food Chem. (2018) 248: 146-54.
[41] Ramirez-Mares MV, Chandra S, de Mejia EG. In vitro chemopreventive activity of Camellia sinensis, Ilex paraguariensis and Ardisia compressa tea extracts and selected polyphenols. Mutat. Res. (2004) 554: 53-65.
[42] Shah S, Gani A, Ahmad M, Shah A, Gani A , Masoodi F. In vitro antioxidant and antiproliferative activity of microwave-extracted green tea and black tea (Camellia sinensis): a comparative study. NutraFoods (2015) 14: 207-15.
[43] Zhao X, Song JL, Kim JD, Lee JS, Park KY. Fermented Pu-erh tea increases in vitro anticancer activities in HT-29 cells and has antiangiogenetic effects on HUVECs. J. Environ. Pathol. Toxicol. Oncol. (2013) 32: 275-88.
[44] Lambert JD, Lee MJ, Diamond L, Ju J, Hong J, Bose M, Newmark HL, Yang CS. Dose-dependent levels of epigallocatechin-3-gallate in human colon cancer cells and mouse plasma and tissues. Drug Metab. Dispos. (2006) 34: 8-11.
[45] Valcic S, Timmermann BN, Alberts DS, Wächter GA, Krutzsch M, Wymer J, Guillén JM. Inhibitory effect of six green tea catechins and caffeine on the growth of four selected human tumor cell lines. Anticancer Drugs (1996) 7: 461-8.
[46] Ravindranath MH, Saravanan TS, Monteclaro CC, Presser N, Ye X, Selvan SR , Brosman S. Epicatechins purified from green tea (Camellia sinensis) differentially suppress growth of gender-dependent human cancer cell lines. Evid. Based Complement. Altern. Med. (2006) 3: 237-47.
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