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Research Article z Available online at http://www.journalcra.com INTERNATIONAL JOURNAL OF CURRENT RESEARCH International Journal of Current Research Vol. 8, Issue, 09, pp.38899-38904, September, 2016 ISSN: 0975-833X RESEARCH ARTICLE IDENTIFICATION AND ESTIMATION OF PHYTOCHEMICALS AND EVALUATION OF ANTICANCER ACTIVITY OF LAGENARIA SICERARIA LEAVES AND FRUIT EXTRACT *Agrawal, R. C. and Shiwa Mishra Department of Research, Priyamvada Birla Cancer Research Institute, J.R.Birla Road, Satna-485005, Madhya Pradesh, India ARTICLE INFO ABSTRACT Article History: Cancer is one of the leading causes of mortality worldwide. The present study was carried out to evaluate the anti-cancer activity of methanolic extract of Lagenaria siceraria on skin Papilloma Received 22nd June, 2016 Received in revised form model in mice. Lagnaria sciceria leaves and fruit extract against 7, 12 - dimethylbenz (a) anthracene 25th July, 2016 (DMBA) induced papillomagenesis in Swiss albino mice was studied. The methanolic extract of Accepted 17th August, 2016 Lagnaria sciceraria was analyzed for chemopreventive activity. It was evaluated by two stage Published online 30th September, 2016 protocol consisting of initiation with a single topical application of a carcinogen (7, 12 - dimethylbenz (a) anthracene (DMBA) followed by a promoter (croton oil) two times in a week were employed. Key words: A significant reduction in tumor incidence, tumor burden and cumulative number of papillomas was observed, along with a significant increase in average latent period in mice treated topically with Papilloma, Lagnaria sciceraria extract as compared to the control group treated with DMBA and croton oil. The DMBA, Phytochemical analysis of methanolic extract of leaves and fruits of Lagnaria sciceraria showed Croton oil, presence of Alkaloids, triterponoids, flavonoids steroid, glycoside, tannin resin and saponin. However Phytochemical, HPTLC. in methanolic extract of fruits of Lagnaria shows presence of all above compounds except alkaloids. The carbohydrate and protein were present in fruit extract which were absent in leaves sample. The above studies revealed information about the anticancer activity of Lagnaria sciceraria extracts. Copyright©2016, Agrawal and Shiwa Mishra. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Citation: Agrawal, R. C. and Shiwa Mishra, 2016. “Identification and estimation of phytochemicals and evaluation of anticancer activity of Lagenaria siceraria leaves and fruit extract”, International Journal of Current Research, 8, (09), 38899-38904. INTRODUCTION throughout India as an edible vegetable. It is a pubescent or trailing herb, with bottle or dumb-bell shaped fruits. Both of its Cancer is one of the most fatal diseases in human population aerial parts and fruits are commonly consumed as vegetable. and one of the most frequent causes of death worldwide. An Traditionally it is used as medicine in India, China, European extremely promising strategy for cancer prevention today is countries, Brazil, Hawaiian island etc. for its cardiotonic, chemoprevention which is defined as the use of synthetic or general tonic, diuretic, antiproliferative properties (Kirtikar and natural agents (alone or combination) to block the development Basu, 2003). Further, antihepatotoxic, analgesic, anti- of cancer in humans. Plants, vegetables and herbs used in the inflammatory, hypolipidemic, antihyperglycemic and folk and traditional medicine have been accepted currently as antioxidant activities of its fruit extract have been reported one of the main sources of cancer chemoprevention in drug (Deshpande et al., 2008; Deshpande et al., 2007; Ghule et al., discovery and development. Plant derived natural products 2006a, b; Shirwaikar and Sreenivasan, 1996). Lagenaria such as flavonoids, terpenoids alkaloids and steroids have siceraria fruits are good source of Vitamin B complex, received considerable attention due to their diverse ascorbic acid, fibers, proteins, cucurbitacins, saponins, pharmacological properties which include cytotoxic and fucosterols and compesterols, polyphenolics, flavones-C- chemopreventive effects (Abdullaev, 2001; Uddin et al., glycoside (Ghule et al., 2006b; Shirwaikar and Sreenivasan, 2003; Koduru et al., 2006; Zahan et al., 2011; Sodde et al., 1996; Krauze-Baranowska and Cisowski, 1994; Duke, 2011; Kundu Sen et al., 2011). The plant, Lagenaria 1999; Sturm and Stuppner, 2000). Methanol extract of its siceraria (Mol.) Stanley from Cucurbitaceae family, popularly leaves was reported the presence of sterols, polyphenolics, known as bottle gourd (English), has wide occurrence flavonoids, saponin, protein and carbohydrates (Shah and Seth, 2010). A novel protein, Lagenin from its seeds was reported *Corresponding author: Agrawal, R. C. antitumor, immunoprotective and antiproliferative properties Department of Research, Priyamvada Birla Cancer Research Institute, (Wang and Ng, 2000). The present investigation was therefore, J.R.Birla Road, Satna-485005, Madhya Pradesh, India. 38900 Agrawal and Shiwa Mishra, Identification and estimation of phytochemicals and evaluation of anticancer activity of Lagenaria siceraria leaves and fruit extract carried out to evaluate anticancer activity of methanol extract maintained under controlled conditions of temperature of L. siceraria leaves and fruit against Skin Papilloma tumor (25±2 °C) and light (12 light: 12 dark). They were fed on model in mice and identification of phytoconstituents present in standard mice feed procured from Golden feeds, New Delhi fruit and leaves of Lagnaria sciceraria. and water was given ad. libitum. One day before the commencement of the experiment, hairs on the interscapular MATERIALS AND METHODS region of the mice were removed using hair removing cream. Phytochemical analysis: Chemicals Preliminary phytochemical screening of the extract was carried 7, 12-dimethylbenz (a) anthracene (DMBA), and Croton oil out using standard methods (Kokate, 1994). were procured from Sigma chemicals Co., St. Louis, U.S.A. and other chemicals were procured locally and were reagent Collection and identification of plant material grade. The plant of L. Siceraria was collected from garden of Procedure agriculture college Sehore (Madhya Pradesh India). The identification of the plant Lagenaria siceraria (family: Experiment was performed as per the method reported by cucutbitacea) was done by Dr. Manoj Tripathi (Voucher Berenblum (1975) and standardized by us (Agrawal et al., Specimen TFRA/AS/S/115), Deendayal Research Institute, 2009). The animals were randomly divided into different Chitrakoot, Satna Madhya Pradesh (India) The project was groups and each group comprised of six animals. Hairs were approved by Institutional Animal Ethical Committee (IAEC). removed with the help of hair removing cream from the dorsal Project no 5 Ref no 670/2251. The experiment was carried out region with proper care in the area of 2cm2 in all the according to the committee for the purpose of control and groups.100 μg DMBA was dissolved in 100μl acetone and was supervision of experiments on animal (CPCSEA) guidelines. given initially and 1% Croton oil was given 2 times a week up to 16 weeks. Skin tumor formation was recorded weekly and Extraction Procedure the papillomas greater than 1mm in diameter and if they persisted two weeks or more were included in to counting of The leaves and fruits of L.Siceraria was dries in shade and total number of papillomas / mouse, Tumor incidence and powdered with mechanical grinder. The powder was passed tumor yield was also calculated. The animals were divided into through sieve no 40 and stored in airtight container for further 8 different groups for each extract as follows: Total No. of studies. About 50 gm of L.Siceraria leaves and fruits powder animals for each group were 6 was kept in petroleum ether to de fat for 1hr and to remove the lipid present in plant material. The powder (L. Siceraria leaves Treatment Groups and fruits) was dried in filter paper. After dry it was poured in separating funnel for extraction using 50% methanol solvent at Group I (Vehicle Control) - 100μl acetone 2 times/ week up room temperature for 24 hr and then filtered. Again the 50% to 16 weeks. methanol solvent was added and allowed to stand for overnight Group II (DMBA alone)-100 μg DMBA was dissolved in and then filtered to concentrated it. The filtrate was kept at 55- 100μl acetone and single application was given. 60’C in water bath. The collected residue was finally Group III (Croton oil alone)-1% Croton oil was applied on transferred into the hot air oven to dry it. About 14 gm crude the skin 2 times a week up to 16 weeks. extract was obtained which was used for the studies. The yield Group IV (DMBA+Croton oil)- 100 μg DMBA was dissolved of methanolic extract was 30%. The determination of Alkaloid in 100μl acetone and single application was given by Harborne (1973) method, Flavonoid by Bohm and Kocipai – afterwards, 1% Croton oil was applied on skin 2 times a Abyazan (1994) method and saponin by Obadani and Ochako week up to 16 weeks. (2001) method was done. Group V L.siceraria leaves extract alone)- L.siceraria leaves Methanolic extract alone was applied (3000mg/kg body HPTLC fingerprint profile wt.) on skin 2 times a week up to 16 weeks. Group VI (L.siceraria fruit extract alone)- L.siceraria fruit HPTLC fingerprinting of methanolic extract of the
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