Antioxidant, anti-cancer and ameliorative activities of Spirulina platensis and pomegranate juice against hepatic damage induced by CCl4

  • Hossam S. EL-BELTAGI King Faisal University, Agricultural Biotechnology Department, College of Agriculture and Food Sciences, P.O. Box 420, Al-Ahsa 31982; Cairo University, Faculty of Agriculture, Biochemistry Department, Gamma st, Giza, P.O. 12613
  • Faten DHAWI King Faisal University, Agricultural Biotechnology Department, College of Agriculture and Food Sciences, P.O. Box 420, Al-Ahsa 31982
  • Ihab ASHOUSH Ain Shams University, Faculty of Agriculture, Food Science Department, Cairo
  • Khaled RAMADAN King Faisal University, Central Laboratories; Ain-Shams University, Faculty of Agriculture, Agricultural Biochemistry Department, Cairo
Keywords: anticancer; antioxidant status; carotenoids; liver injury; pomegranate juice; Spirulina platensis


Due to the excessive impact of synthetic drugs, unravelling and employing safe, natural alternatives are now needed to resolve a number of diseases. In this research, we have evaluated hepatoprotective and antioxidant activities of Spirulina platensis and pomegranate juice in rats against hepatotoxicity induced by carbon tetrachloride (CCl4). Spirulina crude carotenoid extract was screened by UPLC-MS / MS. Activities of liver marker enzymes; measured aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and determined lipid peroxidation and antioxidant status as reduced glutathione (GSH) in liver homogenate. The infusion of CCl4 (2 ml/kg b.wt) greatly increases levels of liver marker enzymes and lipid peroxidation, resulting in depletion of antioxidants. Treatment of Spirulina platensis (Sp), pomegranate juice (Pj)or mixture (PJSP) of Spirulina water extract 10% and pomegranate juice 90% (1 ml/100 g b.wt) to CCl4-disrupted rats resulted in decreased activity of liver marker enzymes, lipid peroxidation with increased antioxidant status. Chromatographic separation showed that ß-carotene is the predominant carotenoid extract. This carotenoid extract was tested for colon carcinoma (HCT-116), liver carcinoma (HepG2) and intestinal carcinoma cell lines (CACO) LC50 for 21.8, 14 and 11.3 ug / ml, respectively. Total phenolic phytochemicals, total carotenoids and total flavonoids were also measured in Spirulina. Our study clearly demonstrates that Spirulina platensis and pomegranate juice had hepatoprotective effect on CCl4-caused hepatotoxicity in rats through its antioxidant activity.


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Abd El-Maksoud AA, Abd El-Ghany IH, El-Beltagi HS, Anankanbil S, Banerijee C, Petersen SV, … Guo Z (2018). Adding functionality to milk-based protein: Preparation, and physicochemical characterization of β-lactoglobulin-phenolic conjugates. Food Chemistry 241:281-289. https://doi.org10.1016/j.foodchem.2017.08.101

Abdel-Salam AM, Ashoush IS, Hamad EM Mousa HM (2015). Potential protective effect of fermented camel milk containing probiotic, prebiotic and synbiotic against lead and cadmium-intoxicated male rats. International Journal of Humanities, Arts, Medicine and Sciences (BEST: IJHAMS) 3(11):131-148.

Abdel-Rahim EA, El-Beltagi HS (2010). Constituents of apple, parsley and lentil edible plants and their therapy treatments for blood picture as well as liver and kidney functions against lipidemic disease. Electronic Journal of Environmental, Agricultural and Food Chemistry 9(6):1117-1127.

Abdel-Rahim EA, El-Beltagi HS, Romela RM (2013). White bean seeds and pomegranate peel and fruit seeds as hypercholesterolemic and hypolipidemic agents in albino rats. Grasas y Aceites 64(1):50-58.

Abe K, Hattori H, Hirano M (2007). Accumulation and antioxidant activity of secondary carotenoids in the aerial microalga Coelastrella striolata var. multistriata. Food Chemistry 100:656-661.

Ahmed SG, Yasser AK, Aziza AE, Sherif RM, Nabila SH, Mosaad AA (2011). Antioxidant activity and hepatoprotective effects of whey protein and Spirulina in rats. Nutrition 27:582-589.

Akladious SA, Mohamed HI (2017). Physiological role of exogenous nitric oxide in improving performance, yield and some biochemical aspects of sunflower plant under zinc stress. Acta Biologica Hungarica 68(1):101-114.

Akram A, Ulfat O, Marwa Y, Mary G, Mary AL, Nazlin H (2016). Antioxidant activity of pomegranate juice and punicalagin. Natural Science 8:235-246.

Al-Olayan EM, El-Khadragy MF, Metwally DM, Abdel Moneim AE (2014). Protective effects of pomegranate (Punica granatum) juice on testes against carbon tetrachloride intoxication in rats. BMC Complementary and Alternative Medicine 14:164.

Aly AA, Mansour MTM, Mohamed HI, Abd-Elsalam KA (2012a). Examination of correlations between several biochemical components and powdery mildew resistance of flax cultivars. Plant Pathology Journal 28(2):149-155.

Asrani SK, Devarbhavi H, Eaton J, Kamath PS. Burden of liver diseases in the world. Journal of Hepatology 70:151-171.

Aviram M, Dornfeld L, Rosenblat M, Volkova N, Kaplan M, Coleman R (2000). Pomegranate juice consumption reduces oxidative stress, atherogenic modifications to LDL, and platelet aggregation: studies in humans and in atherosc­lerotic apolipoprotein E-deficient mice. American Journal of Clinical Nutrition 71:1062-1076.

Balasundram, N, Sundram K, Samman S (2006). Phenolic com­pounds in plants and agri-industrial by-products: anti­oxidant activity, occurrence, and potential uses. Food Chemistry 99:191-203.

Bandyopadhyay U, Das D, Banerjee KR (1999). Reactive oxygen species: Oxidative damage and pathogenesis. Current Science 77:658-66.

Bernal J, Mendiola JA, Ibáñez E, Cifuentes A (2011). Advanced analysis of nutraceuticals. Journal of Pharmaceutical and Biomedical Analysis 55:758-774.

Beutler E, Duron O, Kelly BM (1963). Improved method for the determination of blood glutathione. Journal of Laboratory and Clinical Medicine 61:882-888.

Brand-Williams W, Cuvelier ME, Berset C (1995). Use of a free radical method to evaluate antioxidant activity. LWT - Food Science and Technology 28:25-30.

Cipolatti EP, Remedi RD, Sá CD, Rodrigues AB, Ramos JMG, Burkerta CAV, … Burkert JFD (2019). Use of agroindustrial byproducts as substrate for production of carotenoids with antioxidant potential by wild yeasts. Biocatalysis and Agricultural Biotechnology 20:101208

Chaterrjee TK (2000). Medicinal plants with hepatoprotective properties. Herbal options. Books and Applied Allied (P) Ltd, Calcutta, pp 13.

Czerwonka A, Kaławaj K, Sławińska-Brych A, Lemieszek MK, Bartnik M, Wojtanowski KK, … Rzesk W (2018). Anticancer effect of the water extract of a commercial Spirulina (Arthrospira platensis) product on the human lung cancer A549 cell line. Biomedicine & Pharmacotherapy 106:292-302.

Dartsch PC (2008). Antioxidant potential of selected Spirulina platensis preparations. Phytotherapy Research 22(5):627-633.

de Cruz CR, Lubrano A, Gatlin DM (2018). Evaluation of microalgae concentrates as partial fishmeal replacements for hybrid striped bass Morone sp. Aquaculture 493:130-136.

Djeridane A, Yousfi M, Nadjemi B, Boutassouna D, Stocker P, Vidal N (2006). Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food Chemistry 97:654-660.

El-Baz FK, El-Senousy WM, El-Sayed AB, Kamel MM (2013) In vitro antiviral and antimicrobial activities of Spirulina platensis extract. Journal of Applied Pharmaceutical Science 3:52-56,

El-Beltagi HS, Mohamed HI, Safwat G, Megahed BMH, Gamal M (2018). Evaluation of some chemical constituents, antioxidant, antibacterial and anticancer activities of Beta vulgaris L. root. Fresenius Environmental Bulletin 27(9):6369-6378.

El-Beltagi HS, Mohamed HI, Elmelegy AA, Eldesoky SE, Safwat G (2019a). Phytochemical screening, antimicrobial, antioxidant, anticancer activities and nutritional values of cactus (Opuntia ficus indicia) pulp and peel. Fresenius Environmental Bulletin 28(2A):1534-1551.

El-Beltagi HS, Mohamed HI, Safwat G, Gamal M, Megahed BMH (2019b). Chemical composition and biological activity of Physalis peruviana L. Gesunde Pflanzen 71:113-122.

Faria A, Monteiro R, Mateus N, Azevedo I, Calhau C (2007). Effect of pomegranate (Punica granatum) juice intake on hepatic oxidative stress. European Journal of Nutrition 46:271-278.

Fayyad RJ, Ali ANM, Dwaish AS, Abed Al- Abboodi AK (2019). Anticancer activity of Spirulina platensis methanolic extracts against L20B and MCF7 human cancer cell lines. Plant Archives 19(1):1419-1426.

Feltl L, Pacákováa V, Stulika K, Volka K (2005). Reliability of carotenoid analyses: a review. Current Analytical Chemistry 1:93-102.

Finamore A, Palmery M, Bensehaila S, Peluso I (2017). Antioxidant, immunomodulating, and microbial-modulating activities of the sustainable and ecofriendly Spirulina. Oxidative Medicine and Cellular Longevity 2017:3247528.

García-Martínez D, Rupérez FJ, Ugarte P, Barbas C (2007). Tocopherol fate in plasma and liver of streptozotocin-treated rats that orally received antioxidants and Spirulina extracts. International Journal for Vitamin and Nutrition Research 77(4):263-271.

Hamed MM, Abd El-Mobdy MA, Kamel MT, Mohamed HI, Bayoumi AE (2019). Phytochemical and biological activities of two asteraceae plants Senecio vulgaris and Pluchea dioscoridis L. Pharmacology Online 2:101-121.

Hanson CJ, Bootman MD, Distelhorst CW, Maraldi T, Roderick HL (2008). The cellular concentration of Bcl-2 determines its pro- or anti-apoptotic effect. Cell Calcium 44(3):243-258.

Helmi A, Mohamed HI (2016). Biochemical and ulturasturctural changes of some tomato cultivars to infestation with Aphis gossypii Glover (Hemiptera: Aphididae) at Qalyubiya, Egypt. Gesunde Pflanzen 68:41-50.

Hernandez FYF, Khandual S, Lopez IGR (2017). Cytotoxic effect of Spirulina platensis extracts on human acute leukemia Kasumi-1 and chronic myelogenous leukemia K-562 cell lines. Asian Pacific Journal of Tropical Biomedicine 7(1):9-14.

Ismail MF, Ali DA, Fernando A, Abdraboh ME, Gaur RL, Ibrahim WM, … Ouhtit A (2009). Chemoprevention of rat liver toxicity and carcinogenesis by Spirulina. International Journal of Biological Sciences 5(4):377-387.

Jaiswal V, DerMarderosian A, Porter JR (2010). Anthocyanins and polyphenol oxidase from dried arils of pomegranate (Punica granatum L.). Food Chemistry 118(1):11-16.

Jung MY, Min DB (1991). Effect of quenching mechanisms of carotenoids on the photosensitized oxidation of soybean oil. Journal of the American Oil Chemists Society 68:653-658.

Karan M, Vasisht K, Handa SS (1999). Antihepatotoxic activity of Swertia chirata on carbontetrachloride induced hepatotoxicity in rats. Phytotherapy Research 13:24-30.<24::AID-PTR378>3.0.CO;2-L

Karadeniz A, Cemek M, Simsek N (2009). The effects of Panax ginseng and Spirulina platensis on hepatotoxicity induced by cadmium in rats. Ecotoxicology and Environmental Safety 72(1):231-235.

Keshavan P, Schwemberger SJ, Smith DLH, Babcock GF, Zucker SD (2004). Unconjugated bilirubin induces apoptosis in colon cancer cells by triggering mitochondrial depolarization. International Journal of Cancer 112:433-45.

Kew MC (2002). Epidemiology of hepatocellular carcinoma. Toxicology 181-182:35-38.

Konícková R, Vanková K, Vaníková J, Vánová K, Muchová L, Subhanová I… Vítek L (2014). Anticancer effects of blue-green alga Spirulina platensis, a natural source of bilirubin-like tetrapyrrolic compounds. Annals of Hepatology 13(2):273-283.

Lansky EP (2006). Beware of pomegranates bearing 40% ellagic acid. Journal of Medicinal Food 9:119-22.

Marín L, Miguélez EM, Villar CJ, Lombó F (2015). Bioavailability of dietary polyphenols and gut microbiota metabolism: antimicrobial properties. BioMed Research International 2015:1-18. https://doi.og/10.1155/2015/905215

Małgorzata K, Ewa I, Marek M, Małgorzata E, Katarzyna M, Jodynis-Liebert J (2009). Protective effect of red beetroot against carbon tetrachloride- and n-nitrosodiethylamine-induced oxidative stress in rats. Journal of Agricultural and Food Chemistry 57:2570-2575.

Manal FE (2011). Hepatoprotective role of the pomegranate (Punica granatum) juice on carbon tetrachloride-induced oxidative stress in rats. African Journal of Biological Science 7(1):135-149.

Marzorati S, Schievano A, Idàb A, Verotta L (2020). Carotenoids, chlorophylls and phycocyanin from Spirulina: supercritical CO2 and water extraction methods for added value products cascade. Green Chemistry 22:187-196.

Maskan M (2006). Production of pomegranate (Punica granatum L.) juice concentrate by various heating methods: color degradation and kinetics. Journal of Food Engineering 72:218-224.

Matthaiou CM, Goutzourelas N, Stagos D, Sarafoglou E, Jamurtas A, Koulocheri SD, … Kouretas D (2014). Pomegranate juice consumption increases GSH levels and re­duces lipid and protein oxidation in human blood. Food and Chemical Toxicology 73:1-6.

Mendiola JA, Marín FR, Hernandez S, Arredondo BO, Señoráns FJ, Ibañez E, Reglero G (2005). Characterization via liquid chromatography coupled to diode array detector and tandem mass spectrometry of supercritical fluid antioxidant extracts of Spirulina platensis microalga. Journal of Separation Science 28:1031-1038.

Mohamed HI, Akladious SA (2017). Changes in antioxidants potential, secondary metabolites and plant hormones induced by different fungicides treatment in cotton plants. Pesticide Physiology and Biochemistry 142:117-122.

Nagao A (2014). Bioavailability of dietary carotenoids: intestinal absorption and metabolism. Japan Agricultural Research Quarterly 48(4):385-391.

Nakamura H, Uetani Y, Komura M, Takada S, Sano K, Matsuo T (1987). Inhibitory action of bilirubin on superoxide production by polymorphonuclear leukocytes. Biology of the Neonate 52:273-278.

Nirwane A, Patil R (2012). Effect of Punica granatum seeds on doxorubicin-induced hepatotoxicity in Wistar Rats. Journal of Pharmacy Research 5:4625-4628.

Ohkawa H, Ohishi N, Yagi K (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95:351-358.

Ollinger R, Kogler P, Troppmair J, Hermann M, Wurm M, Drasche A, … Margreiter R (2007). Bilirubin inhibits tumor cell growth via activation of ERK. Cell Cycle 6:3078-3085.

Panek J, Fantova L, Trojakova L, Reblova Z, Pokorny J (2001). Effectiveness of natural antioxidants in sunflower oil. Special Publication– Royal Society of Chemistry 269:371-374.

Prabakaran G, Sampathkumar P, Kavisri M, Moovendhan M (2020). Extraction and characterization of phycocyanin from Spirulina platensis and evaluation of its anticancer, antidiabetic and antiinflammatory effect. International Journal of Biological Macromolecules 153:256-263.

Reeves PG, Nielsen FH, Fahey Jr, GC (1993). AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition ad hoc writing committee on the reformulation of the AIN-76A rodent diet. Journal of Nutrition 123(11):1939-1951.

Reitman S, Frankel S (1957). A colorimetric method of the determination of plasma glutamic oxaloacetic and glutamic pyruvic transaminases. American Journal of Clinical Pathology 28:56-63.

Remziye AK, Sait P, Ahmet C, Nuray B, Saadet DS (2013). Protective effect of Spirulina platensis enriched in phenolic compounds against hepatotoxicity induced by CCl4. Food Chemistry 141:1972-1979.

Rechter S, Konig T, Auerochs S, Thulke S, Walter H, Dornenburg HD, Walter C, Marschall M (2006). Antiviral activity of Arthrospira-derived spirulan-like substances. Antiviral Research 72:197-206,

Rosenblat M, Hayek T, Aviram M (2006). Antioxidative effects of pomegranate juice (PJ) consumption by diabetic pa­tients on serum and on macrophages. Atherosclerosis 187:363-371.

Rouanet JM, Kelly D, Daniele DR, Cyril A, Gina B, Jean-Paul C, … Alan C (2010). Berry juices, teas, antioxidants and the prevention of atherosclerosis in hamsters. Food Chemistry 118:266-271.

Sadia H, Akter QS, Afroz R, Siddiqua T (2016). Effect of Punica granatum (Pomegranate) on serum ALT and AST in carbon tetrachloride induced liver damage in Wistar albino rats. Journal of Bangladesh Society of Physiologist 11(1):23-28.

Sallam AE, Mansour AT, Srour TM, Goda AMA (2017). Effects of different carotenoid supplementation sources with or without sodium taurocholate on growth, feed utilization, carotenoid content and antioxidant status in fry of the European seabass, Dicentrarchus labrax. Aquaculture Research 48(7):3848-3858.

Sanoner P, Guyot S, Marnet N, Molle D, Drilleau JF (1999). Polyphenol profiles of French cider apple varieties (Malus domestica sp.). Journal of Agricultural and Food Chemistry 47:4847-4853.

SAS (2003). SAS/ Stat Users Guide: Statistics, System for Windows, version 4.10 (release 8.01 TS level 01M0), SAS Inst, Inc. Cary, North Carolina, USA.

Seeram NP, Adams LS, Henning SM (2005). In vitro antiproli­ferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. Journal of Nutritional Biochemistry 16:360-367.

Serban M-C, Sahebkar A, Dragan S, Stoichescu-Hogea G, Ursoniu S, Andrica F, Banach M (2016). A systematic review and meta-analysis of the impact of Spirulina supplementation on plasma lipid concentrations. Clinical Nutrition 35(4):842-851.

Sies H (1999). Glutathione and its role in cellular functions. Free Radical Biology and Medicine 27:916-21.

Sharma MK, Sharma A, Kumar A, Kumar M (2007). Spirulina fusiforms provides protection against mercuric chloride induced oxidative stress in Swiss albino mice. Food and Chemical Toxicology 45:2412-2419.

Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, … Boyd MR (1990). New colorimetric cytotoxicity assay for anticancer-drug screening. Journal of The National Cancer Institute 82(13):1107-1112.

Soni RA, Sudhakar K, Rana R (2017). Spirulina-from growth to nutritional product: A review. Trends in Food Science and Technology 69:157-171.

Subagio A, Morita N, Sawada S (1996). Carotenoids and their fatty acid esters in banana peel. Journal of Nutritional Science and Vitaminology 42:553-566.

Subramonium A, Pushpangadan P (1999). Development of phytomedicines for liver diseases. Indian Journal Pharmacology 31:166-175.

Surh YJ (1999). Molecular mechanisms of chemopreventive effects of selected dietary and medical phenolic substances. Mutation Research 428(1-2):305-327.

Vadiraja BB, Gaikwad NW, Madhyastha KM (1998). Hepato-protective effect of C- Phycocyanin: protection for carbon tetrachloride and pulegeon mediated hepato-toxicity in rats. Biochemical and Biophysical Research Communications 249(2):428-431.

Ward FM, Daly MJ (1999). Hepatic Disease. In: Clinical Pharmacy and Therapeutics Walker R, Edwards C (Eds). Churchill Livingstone, New York, pp 195-212.

Wu LC, Ho JA, Shieh MC, Lu IW (2005). Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts. Journal of Agricultural and Food Chemistry 53(10):4207-4212.

Yin C, Daoust K, Young A, Tebbs E, Harper D (2017). Tackling community undernutrition at Lake Bogoria, Kenya: the potential of Spirulina (Arthrospira fusiformis) as a food supplement. African Journal of Food, Agriculture, Nutrition and Development 17:11603-11615.

Yoshikawa N, Belay A (2008). Single-laboratory validation of a method for the determination of c-phycocyanin and allophycocyanin in Spirulina (Arthrospira) supplements and raw materials by spectrophotometry. Journal of AOAC International 91(3):524-529.

Zhang P, Omaye ST (2001). Antioxidant and prooxidant roles for beta-carotene, alpha-tocopherol and ascorbic acid in human lung cells. Toxicology in Vitro 15(1):13-24.

How to Cite
EL-BELTAGI, H. S., DHAWI, F., ASHOUSH, I. S., & RAMADAN, K. (2020). Antioxidant, anti-cancer and ameliorative activities of Spirulina platensis and pomegranate juice against hepatic damage induced by CCl4. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 48(4), 1941-1956.
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