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Amaranthus Spinosus Leaf Extracts and Its Anti-Inflamatory Effects on Cancer S

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Amaranthus Spinosus Leaf Extracts and Its Anti-Inflamatory Effects on Cancer S Dinesh et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online) Amaranthus spinosus leaf extracts and its anti-inflamatory effects on cancer S. Rajasekaran2, M.G. Dinesh1*,Chandarasekharam Kansrajh3, Fida Hussain Ahmed Baig2 1. Department of Biotechnology, Thanthai Hans Roever College, Perambalur, 2. University of East London (School of Health and Bio-Sciences) 3. Madras Christian college, Chennai *Corresponding author: [email protected] ABSTRACT The fruits and vegetables generate phytochemicals by its nature such as flavonoids and antioxidants which can reduce the oxidative stress and lower risk of chronic sustainment like cancer. The present study was to investigate and evaluate the antitumor capacity and chemoprotective effects of Amaranthus Spinosus leaves. The methanol extracts of Amaranthus Spinosus leaves tested with different doses for different cell lines such as breast, colorectal, liver and normal cell lines. This was observed that the methanol leaf extract at different doses causes a decrease significant level of tumor development and viable cell count. Percentage of leaf Methanol extracts HEP G2, MCF-7 and HT-29 cells shows IC 50 values of less than 30ug/ml. Amaranthus Spinousus leaf extracts shown significant membrane stability property which shows better anti-inflammatory responses. FACS used to analysis the inhibition of cell cycle progressionsby using PI Staining for HEp G2 Cells. The results suggest that the methanol extracts of Amaranthus spinosus leaves reveals significant antitumor effects in cancers of breast, colorectal, liver and normal cell lines. Further studies are required to elucidate the precise molecular mechanisms and targets for cell growth inhibition. Key words: Amaranthus spinosus, Anti-inflammatory, Cancer INTRODUCTION remedy for treating human diseases because they contain numerous active constituents of immense During the last decades, chemists and therapeutic value. Even now, approximately 80% of biologists havebeen intensively investigating tropical the third world population is almost entirely and subtropicalplants species with potential medicinal dependent on traditional medicines for maintaining properties inorder to assess the feasibility of general health and combating many diseases Plants developing natural, sustainable, and affordable drugs and plant products both as extracts and derived and cosmetics (AkéAssi and Guinko, 1991). compoundsare known to be effective and versatile Medicinal plant exhibits antibacterial activitysince chemo preventive agents various types of cancers. The they contain innumerable biologically activechemical plant A.spinosus Linn (Amaranthaceae) is an annual constituents. The use of plant preparation asfoodstuff, herb found in throughout India and also many tropical insecticides, CNS active, cardio active, antitumorand countries. Traditionally the plant is used to treat antimicrobial agents are some examples ofimmense various diseases. Although the leaves part are used as chemical diversity in plants, which are as old a laxative and an applied as an emollient poultice to asmankind (Al-Sereiti, 1999).Plants produce a diverse abscesses, boils and burns and reported as range of bioactive molecules, making them a rich antimalarial,antioxidant and anti-hepatotoxic actions. source of different types ofmedicines. A rich heritage The presence of spinoside, new coumaroyl flavone of knowledge to preventive and curative medicines glycoside lectins, betacyanins and phenolic was available in ancientscholastic works included in compounds has been reported. the Atharva Veda, Charaka, Sushruta etc. Over 50% of all modern clinicaldrugs are of natural product Amaranthus spinosus Linn. (Amaranthaceae) origin (Stuffness, 1982) and natural products play an is commonly known as “KateWali Chaulai important rolein drug development programs in the (Kanatabhajii)” in Hindi, and is used as avegetable pharmaceutical industry (Baker, 1995). and cultivated throughout India, Sri Lanka, andmany other tropical countries. In Ayurveda (Indian Herbal drugs havegained importance in recent traditionalsystem of medicine) the plant is used as a years because of their efficacy and cost effectiveness. digestive,laxative, diuretic, stomachic, and antipyretic, About 80% of individuals from developed countries to improveappetite, biliousness, blood diseases, use traditional medicine, derived from medicinal burning sensation,leprosy, bronchitis, rat bite, piles, plants. Medicinal plants have been used as an and leucorrhea, whilethe boiled leaves and root are exemplary source for centuries as an alternative given to children as a laxative,emollient, and poultice IJRPB 2(1) www.ijrpb.com January-February 2014 Page 1058 Dinesh et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online) for abscesses, boils, and burns (Kirtikar & Basu, Leaves were washed with sterile water, dried in shade, 2001). The leaves are usedto treat rheumatic pain, finely powdered & stored in air tight bottles. stomach-ache, eczema, gastroenteritis,gallbladder Preparation of plant extract: 25 g of air-dried inflammation, boils, abscesses, snakebite, colic powder of Amaranthus Spinosus leaves was immersed menorrhagia, and arthritis. Amaranthus spinosus is in 100 mL of Methanol in a conical flask. It was also used as an anti-inflammatory, antimalarial, incubated at room temperature for 48 hour at 150 rpm antibacterial,antimicrobial, antidiuretic, and antiviral in an orbital shaker. The suspension was filtered and agent and in hepatic disorders (Olajide, 2004; concentrated to dryness at 40oC in hot air oven. The Stintzing, 2004 ). extract was dissolved in 0.25% Dimethyl Sulphoxide A. spinosus were identified as amaranthine, (DMSO, Merck) to a concentration of 100mg/mL. Isoamaranthine, Hydroxycinnamates, Quercetin, and Cell lines: Cells lines representing the most common Kaempferol Glycosides (Hilou, 2006; Rastogi & human cancers (WHO, 2006) were obtained from the Mehrotra, 1999; Srinivasan, 2005; Stintzing, 2004). King Institute of Preventive Medicine, Chennai. These The plant has a high concentration of antioxidant included the breast adenocarcinoma (MCF-7), the components, high nutritive value due to the presence human Liver Adenocarcinoma (HEP G2) , Human of fiber and proteins, and a high concentration of Laryngeal cancer cell lines (HEp2) , African Green essential amino acids, especially lysine (Cao, 1996; Monkey Kidney Cell lines (VERO ) and the colon Odhav, 2007). adenocarcinoma (HT-29). Scientific studies indicate that the promising Phytochemical analysis of plant extracts: phytochemicals can be developed from the medicinal Phytochemical screening of the extracts was carried plants for many health problems. Moreover, the herbal out according to the methods described by (Jacob et drugs have gained importance in recent years because al., 2011) for the detection of active components like of their efficacy and cost effectiveness. These drugs saponins, tannins, alkaloids, phlobatanins, etc. are invariably single plant extracts or fractions thereof or mixtures of fractions/extracts from different plants, a) Alkaloids- 1 ml of 1% HCl was added to 3 ml of which have been carefully, standardized for their the extract in a test tube. The mixture was then heated safety and efficacy. Amaranthus spinosus Linn. for 20 min, cooled and filtered about 2 drops of (Amaranthaceae) commonly known as Spiny Mayer’s reagent to1 ml of the extract. A creamy amaranth or Pig weed, is an annual or perennial herb, precipitate was an indication of the presence of native to tropical America and found throughout India alkaloids. as a weed in cultivated as well as fallow lands. b) Tannins- 1 ml of freshly prepared 10% KOH was Though whole plant is used as laxative, the root are added to 1 ml of the extract. A dirty white precipitate regarded as highly specific for colic by Hindu showed the presence of tannins. physicians (Sivarajan & Balachandran 1994) and in Madagascar they are considered as laxative (Kirtikar d) Saponins- Frothing test: 2 ml of the extract was & Basu 1987).Traditionally boiled leaves and roots of vigorously shaken in the test tube for 2 min. No Amaranthus spinosus are given to children as laxative. frothing was observed. However the drug is also used traditionally as diuretic, e) Flavonoids- 1 ml of 10% NaOH was added to 3 ml antidiabetic, antipyretic, anti-snake venum, of the extract antileprotic, and anti-gonorrheal (Kirtikar & Basu 1987). In Malaysia, Amaranthus spinosus is used as Cell viability assay: The antitumor assay was an expectorant and to relieve breathing in acute performed on Human laryngeal epithiloma cells bronchitis. Some tribes in India apply A. spinosus to (HEp2) obtained from King Institute of Preventive induce abortion. The A. spinosus is reported for its Medicine, Chennai, India with non-toxic dose of the anti-inflammatory properties, effect on hematology, plant extract and its dilutions. The cells were grown in immunomodulatory activity, antiandrogenic activity, a 24-wellplate in Eagle’s Minimum Essential Medium anthelmintic properties and effect on biochemical (Hi Media) supplemented with 10% fetal bovine changes in epididymis. serum (Gibco Laboratories) and antibiotics (streptomycin, penicillin-G, kanamycin, Amphotericin MATERIALS AND METHODS B). About 1 mL cell suspension (105 cells/mL) was Plant material: Amaranthus Spinosus leaves were seeded in each well and incubated at 37º C for 48 hour obtained
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