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Toxicity Studies on Leaf Extracts of Alternanthera Brasiliana (L.) Kuntze and Alternanthera Bettzickiana (Regel) Voss

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Toxicity Studies on Leaf Extracts of Alternanthera Brasiliana (L.) Kuntze and Alternanthera Bettzickiana (Regel) Voss Journal of Applied Pharmaceutical Science Vol. 8(10), pp 082-089, October, 2018 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2018.81011 ISSN 2231-3354 Toxicity Studies on Leaf Extracts of Alternanthera brasiliana (L.) Kuntze and Alternanthera bettzickiana (Regel) Voss Kasthuri O R1*, Ramesh B2 1Assistant Professor, Department of Biochemistry, Navarasam Arts and Science College for Women, Arachalur, Erode 638101, Tamilnadu, India. 2Associate Professor, Department of Biochemistry, PSG College of Arts and Science, Coimbatore 641014, Tamilnadu, India. ARTICLE INFO ABSTRACT Article history: Herbal medicine is the source for the search of many novel therapeutic compounds in developing countries. Before Received on: 03/06/2018 used as medicine, drugs from plant origin must be ensured as safe. Objective: The present work focused to study the Accepted on: 13/08/2018 in vivo and in vitro toxicity effects of hydroethanolic leaf extracts of Alternanthera brasiliana (A. brasiliana) and Available online: 31/10/2018 Alternanthera bettzickiana (A. bettzickiana). Methods: Sub-acute toxicity studies of hydroethanolic leaf extracts of A. brasiliana and A. bettzickiana were carried out in vivo on mice. 11 groups of albino mice were treated with five different doses of the hydroethanolic leaf extracts of A. brasiliana and A. bettzickiana orally for 14 days. General appearance Key words: and behavior were observed for 14 consecutive days. Effect on hematological parameters and histopathological Cytotoxicity, LD , 50 changes were also monitored. Cytotoxicity was assessed by observing its toxicity in vitro against DLA cell line by hematology, MTT assay, using MTT assay. Results: Sub-acute toxicity studies results showed that up to the tested dose of 2000 mg/kg bwt in histopathology. both extract treatments, throughout the 14 days of treatment, the extract does not produced any toxicity symptoms. MTT assay indicated that both leaf extracts exhibited significant concentration-dependent in vitro cytotoxic activity against DLA cell line. Hydroethanolic leaf extract of A. bettzickiana exhibited more potent cytotoxic effects on DLA cells than A. brasiliana extract. Conclusion: No toxicity related symptoms were observed in different doses of leaf extract treated mice groups. So the LD50 values of the tested leaf extracts were more than 2000 mg/kg bwt. INTRODUCTION Since 1950, a vast number of plant-derived agents Cancer is considered to be one of the most dreaded are used to treat cancer (Kinzler and Vogelstein, 2002). Plants diseases. Uncontrolled cell proliferation or metastasis of abnormal contain a large profile of secondary metabolites that are mainly cells in the body results in cancer (Kaufman and Chabner, 1996). responsible for their cytotoxic activity. In the development of Cancer is found to be one of the major factor causing mortality. anti-cancer agents, the isolation of vincristine and vinblastine More than one-third of the world’s population is affected by cancer. from vinca and podophyllotoxins from Podophyllumhexandrum It accounts for more than 20% of all deaths. Causes of cancer are considered as milestones (Newman et al., 2003). Various include tobacco, viral infection, chemicals, radiation, environmental traditional herbal medicines are used by a majority of the people factors, and dietary factors (Lemkebthomas et al., 2008). At in developing countries to treat a number of diseases and ailments present, the common treatment strategies in cancer are the use of (Liu, 2011). Herbal based medications are often considered to chemotherapeutic agents, surgery, and radiation. These treatments be safe as they are natural and free from side effects Lopes( et are not fully effective and cause many side effects. Hence, there is al., 2000). The increase in the popularity of herbal remedies a great interest to develop safe and low cost anticancer agents from and the limited number of scientific works on their safety and natural sources (Moongkarndi et al., 2004). efficacy, toxicity and adverse effects related to herbal remedies are widely recognized (Saad et al., 2006). There are growing *Corresponding Author evidence that support the toxicity of herbal medicines towards Kasthuri OR, Assistant Professor of Biochemistry, W/O Baraneedaran, their users. Though various studies of the pharmacological 138 Paruvachi, Bhavani Erode(dt)-638312, Tamilnadu, India. potential of medicinal plants have been carried out in the past, E-mail: kasthure @ gmail.com © 2018 Kasthuri OR and Ramesh B. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-Share- AlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/). Kasthuri and Ramesh / Journal of Applied Pharmaceutical Science 8 (10); 2018: 082-089 083 works investigating their potential toxicities are very limited 2000 mg/kg bwt, orally once a day for 14 days. 11 groups of mice (Wojcikowski et al., 2004). were used to study the acute toxicity and each group consisting of No drug should be used clinically without its clinical 3 mice as per the Institutional Animal Ethical Committee (IAEC) trials and toxicity studies (Anisuzzaman et al., 2001). Sub-acute Proposal No: SVCOP/IAEC/013/2016-17 dt 10.03.2017. oral toxicity studies of herbal medicines are essential to identify the safety and the determination of dose level that could be used EXPERIMENTAL GROUPS subsequently. It also helps in the investigation of the therapeutic Group I: Normal control mice index of drugs and xenobiotics (Rang et al., 2001). Group II: Mice fed orally with 100 mg/kg bwt of A. A. brasiliana and A. bettzickiana were herbaceous plants brasiliana leaf extract for 14 days. belonging to the family Amaranthaceae. Amaranthaceae family Group III: Mice fed orally with 250 mg/kg bwt of A. consists of 64 genera and about 800 species, represented by herbs brasiliana leaf extract for 14 days. and shrubs. This cosmopolitan family is most abundant in tropical Group IV: Mice fed orally with 500 mg/kg bwt of A. regions of Africa, India, and America (Hussien, 2005). The leaves brasiliana leaf extract for 14 days. of A. brasiliana are most widely used for therapeutic applications. Group V: Mice fed orally with 1000 mg/kg bwt of A. The phytochemical constituents reported in this plant are flavonol brasiliana leaf extract for 14 days. glycosides (3-O-robinobioside derivatives of kaempferol and Group VI: Mice fed orally with 2000 mg/kg bwt of A. quercetin), vitamins (riboflavin and niacin), betacianin and steroids brasiliana leaf extract for 14 days. such as β-sitosterol (Kumar et al., 2011). The whole plant of A. Group VII: Mice fed orally with 100 mg/kg bwt of A. bettzickiana was useful in nourishing and purifying the blood, as a bettzickiana leaf extract for 14 days. soft laxative, as antipyretic, as a galactagogue and also had wound Group VIII: Mice fed orally with 250 mg/kg bwt of A. healing property. Studies in the liver of ovariectomized mice bettzickiana leaf extract for 14 days. showed the potential of A. bettzickiana in improving superoxide Group IX: Mice fed orally with 500 mg/kg bwt of A. dismutase and catalase activities (Suphanthip et al., 2013). bettzickiana leaf extract for 14 days. In our preliminary study, phytochemical constituents and Group X: Mice fed orally with 1000 mg/kg bwt of A. in vitro antioxidant potentials of A. brasiliana and A. bettzickiana bettzickiana leaf extract for 14 days. were carried out (Kasthuri and Ramesh, 2018). The current work, Group XI: Mice fed orally with 2000 mg/kg bwt of A. sub-acute toxicity study is formulated to find out the safe dose level bettzickiana leaf extract for 14 days. and LD50 values of hydroethanolic leaf extracts of A. brasiliana On the evaluation of acute toxicity, all groups of mice and A. bettzickiana in swiss albino mice model. The leaf extracts treated separately with hydroethanolic leaf extracts of A. brasiliana were also screened in vitro for cytotoxic activity against DLA cell and A. bettzickiana were monitored during the entire study line using MTT assay. period for the signs and symptoms of toxicity and/or mortality, behavioral alterations, food and water intake and changes in body MATERIALS AND METHODS weight. Blood samples were collected after 24 hrs of the last dose of hydroethanolic leaf extracts for the analysis of hematological Plant materials and extraction parameters. On day 14, the mice in each group were sacrificed by The healthy leaves of A. brasiliana and A. bettzickiana cervical dislocation, a small portion of the liver sample was fixed were collected from SKM Siddha and Ayurvedha, Erode. The plants in 10% formalin for the analysis of histopathological architecture. were identified and authenticated at Botanical Survey of India, Coimbatore with voucher number BSI/SRC/5/23/2015/Tech/100 Estimation of hematological profile for Alternanthera brasiliana (L). Kuntze – AMARANTHACEAE The hematological parameters such as hemoglobin, and BSI/SRC/5/23/2015/Tech/101 for Alternanthera bettzickiana PCV, WBC, RBC, and platelets were estimated. The whole (Regel) Voss – AMARANTHACEAE. The leaves of A. brasiliana blood sample was analyzed using SYSMEX Xs – 800i automatic and A. bettzickiana were separately washed, shade dried and was hematology analyzer. coarsely powdered using a mechanical grinder. The shade dried coarsely powdered leaf samples of (500 g) A. brasiliana and In vitro cytotoxicity - MTT assay A. bettzickiana were extracted with hydroethanol by using hot MTT assay was employed to study the in vitro continuous percolation process (Soxhlet). The leaf extracts were cytotoxicity of hydroethanolic leaf extracts of A. brasiliana and concentrated by using a rotary vacuum evaporator (Buchi) at 50°C, A. bettzickiana. dried in a vacuum dessicator and stored at –20°C till further use. MTT assay Sub-acute toxicity studies MTT[3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazol- The in vivo sub-acute toxicity studies of Alternanthera ium Bromide] assay has been used widely for testing the in vitro leaf extracts were conducted according to OECD guideline 423 chemosensitivity of tumor cell lines.
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