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Hepatoprotective, Anti-Inflammatory and Antioxidant Activities Of View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Bulletin of Faculty of Pharmacy, Cairo University (2016) 54, 209–217 Cairo University Bulletin of Faculty of Pharmacy, Cairo University www.elsevier.com/locate/bfopcu www.sciencedirect.com ORIGINAL ARTICLE Hepatoprotective, anti-inflammatory and antioxidant activities of Flacourtia montana J. Grah leaf extract in male Wistar rats Chinchu Joshy, P.A. Thahimon, R. Arun Kumar, Betty Carla, Christudas Sunil * St. Joseph’s College of Pharmacy, Cherthala, Kerala, India Received 16 February 2016; revised 6 May 2016; accepted 5 June 2016 Available online 21 June 2016 KEYWORDS Abstract Flacourtia montana and its related species have been used traditionally for the treatment Hepatoprotective; of various diseases. The present study evaluated the hepatoprotective, anti-inflammatory and Anti-inflammatory; antioxidant activities of F. montana methanolic extract. The hepatoprotective effect of F. montana Antioxidant; was evaluated against paracetamol induced hepatotoxicity in Wistar rats. Administration of parac- Flacourtia montana; etamol (2 g/kg) showed a significant biochemical and histological deterioration in the liver of exper- Carrageenan imental animals. Pretreatment with F. montana (200 and 400 mg/kg b.wt. p.o) significantly (P 6 0.001) reduced the elevated levels of serum enzymes like serum glutamic-oxaloacetic transam- inase (AST), serum glutamic-pyruvic transaminase (ALT), alkaline phosphatase (ALP) and reversed the hepatic damage in the liver which evidenced the hepatoprotective activity. The anti- inflammatory activity of F. montana was evaluated by carrageenan-induced paw edema and cotton pellet-induced granuloma models. F. montana (200 and 400 mg/kg) showed a significant (P 6 0.001) reduction in rat paw edema with 76.39% and 80.32%, respectively induced by carrageenan against the reference anti-inflammatory drug ibuprofen (10 mg/kg) (83.10%). Oral administration of F. montana (200 and 400 mg/kg) also significantly (P 6 0.001) reduced the granuloma mass formation in cotton pellet granuloma method. The reducing power and hydrogen peroxide radical scavenging were increased at increasing doses of F. montana. The results of the present study demonstrate that the methanolic extract of F. montana possess hepatoprotective, anti-inflammatory and antioxidant activities. Ó 2016 Publishing services provided by Elsevier B.V. on behalf of Faculty of Pharmacy, Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc- nd/4.0/). 1. Introduction * Corresponding author at: Department of Pharmacology, St. Joseph’s College of Pharmacy, Cherthala, Kerala, India. Tel.: +91 Hepatotoxicity refers to liver dysfunction or liver damage that 9789688425. is associated with an overload of drugs or xenobiotics such as E-mail address: [email protected] (C. Sunil). acetaminophen, cadmium chloride, ethanol, carbon tetrachlo- 1,2 Peer review under responsibility of Faculty of Pharmacy, Cairo ride (CCl4) and allyl alcohols. The chemicals that cause liver University. injury are called hepatotoxins or hepatotoxicants. Hepatotox- http://dx.doi.org/10.1016/j.bfopcu.2016.06.001 1110-0931 Ó 2016 Publishing services provided by Elsevier B.V. on behalf of Faculty of Pharmacy, Cairo University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). 210 C. Joshy et al. icants are exogenous compounds of clinical relevance and may has not been studied extensively both for its pharmacological include overdoses of certain medicinal drugs, industrial chem- as well as phytochemical properties. But related species of F. icals, and natural chemicals like microcystins, herbal remedies montana, Flacourtia jangomas, Flacourtia sepiaria, Flacourtia and dietary supplements.3,4 Certain drugs may cause liver inermis and Flacourtia ramontchi have been reported with var- injury when introduced even within the therapeutic ranges. ious pharmacological activities like antibacterial, antidiabetic, Hepatotoxicity may result not only from direct toxicity of anti-inflammatory and hepatoprotective.19–21 Hence this study the primary compound but also from a reactive metabolite was designed to evaluate the hepatoprotective, anti- or from an immunologically-mediated response affecting hep- inflammatory and antioxidant activities of F. montana leaves atocytes, biliary epithelial cells and/or liver vasculature.5,6 methanolic extract. Hepatotoxic response is expressed in the form of characteristic patterns of cytolethality in specific zones of the acinus. Its 2. Materials and methods related symptoms may include jaundice or icterus appearance causing yellowing of the skin, eyes and mucous membranes 2.1. Plant material due to high level of bilirubin in the extracellular fluid, pruritus, severe abdominal pain, nausea or vomiting, weakness, severe fatigue, continuous bleeding, skin rashes, generalized itching, The plant material was collected during the month of January, swelling of the feet and or legs, abnormal and rapid weight 2015, from Alappuzha district in Kerala, India. The plant was gain in a short period of time, dark urine and light color stool. taxonomically identified and authenticated by Dr. Kiranraj M. The liver disorders are one of the serious health problems, S. Assistant Professor of Botany S. N College, Cherthala. A throughout the world. Hepatoprotection or antihepatotoxicity voucher specimen was deposited at the Herbarium of Depart- is the ability to prevent damage to the liver.7,8 Hepatoprotec- ment of Botany with accession No. SNCCH089 for future tive effect was studied against chemicals and drug induced hep- reference. atotoxicity in rats. Alcohol, CCl4, galactosamine, paracetamol, isoniazid rifampicin, antibiotics, peroxidised oil, aflatoxin have 2.2. Preparation of methanolic extract of F. montana leaves been used as hepatic agents.9 Inflammation is a part of the complex biological response Leaves were washed thoroughly with water to remove the soil of body tissues to harmful stimuli, such as pathogens, dam- particles, shade dried and grounded. 1 kg of powder was aged cells, or irritants.10 It is a protective response that soaked in methanol sequentially for 72 h at room temperature. involves immune cells, blood vessels, and molecular mediators. Plant debris were removed from the extract by filtration and The purpose of inflammation is to eliminate the initial cause of defatted with petroleum ether and the filtrate was concentrated cell injury, clear out necrotic cells and tissues damaged from under reduced pressure using a rotary evaporator at 40 °C the original insult and the inflammatory process, and to initi- until extraction solvent was completely dried. The yield of ate tissue repair.11 Inflammation is beneficial when needed, methanol extract was 6.1 w/v. The extract was stored in the but it is disastrous when chronic. Chronic inflammation gener- refrigerator at 4 °C for further use. The extract was dissolved ates a constant supply of free radicals that overwhelm our in normal saline and used for animal studies. antioxidant defenses and damage DNA, aging us and causing disease of every description. Inflammation can also be defined 2.3. Experimental animals as body reaction against injury or irritant.12 Microorganisms, physical agents, chemicals, inappropriate immunological Healthy male Wistar rats (150–180 g) were obtained from the responses, and tissue death are the factors that can stimulate Animal house of Gov. Veterinary College, Mannuthy. They inflammation. Infectious agents such as viruses and bacteria were housed in polypropylene cages in a conducive environ- are some of the most common stimuli of inflammation.13 mental situation i.e. temperature (22 ± 2 °C), humidity (45 Since the beginning of human civilization, medicinal plants ±5°C) and 12 h of dark and light cycle. The animals were have been used by mankind for its therapeutic value. Nature fed with normal laboratory chow standard pellet diet. The ani- has been a source of medicinal agents for thousands of years mals were allowed to acclimatize for 7 days before commenc- and an impressive number of modern drugs have been isolated ing the experiments. All the studies were conducted in from natural sources.14 The plant-based, traditional medicine accordance with the animal ethics committee of the institute systems continue to play an essential role in health care, with (IAEC, proposal number SJCP/IAEC/21/03/15/01) St. about 80% of the world’s inhabitants relying mainly on tradi- Joseph’s college of pharmacy, Cherthala. tional medicines for their primary health care.15 The medicinal properties of plants could be based on the antioxidant, antimi- crobial antipyretic effects of the phytochemicals in them.16 2.4. Acute toxicity study According to World Health Organization, medicinal plants would be the best source to obtain a variety of drugs. There- To evaluate the toxicity of F. montana extract the acute toxic- fore, such plants should be investigated to better understand ity study was performed based on Irwin test.22 Four groups of their properties, safety and efficacy.17 fasted healthy rats (six per group) were orally administered the Flacourtia montana J. Grah is a thorny tree that belongs to extract at a dose of 1–4 g/kg; the control group was given dis- the family flacourtiaceae. They are found in the evergreen and tilled water. The rats were observed for 1 h continuously and semi-evergreen
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