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Anticarcinogenic Potential of Ethanol Extract of Indigofera Cordifolia Roth

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Anticarcinogenic Potential of Ethanol Extract of Indigofera Cordifolia Roth Brazilian Journal of Biological Sciences, 2017, v. 4, No. 7, p. 53-66. ISSN 2358-2731 http://dx.doi.org/10.21472/bjbs.040707 Anticarcinogenic potential of ethanol extract of Indigofera cordifolia Roth. (Fabales: Fabaceae) on diethylnitrosamine induced hepatocarcinogenesis in rats Raju Senthil Kumar¹,*, Gopal Murugananthan¹ and Balasubramanian Rajkapoor² ¹Natural Products Laboratory. Swamy Vivekanandha College of Pharmacy. Tiruchengode. Tamilnadu. India. *Email: [email protected]. ²Department of Pharmacology. Faculty of Medicine. Sebha University. Sebha. Libya. Abstract. Indigofera cordifolia Roth. (Fabales: Fabaceae) has been widely used in Indian system of medicine to treat various disorders. Received Earlier studies showed that I. cordifolia (EIC) possess antioxidant, May 17, 2017 free radical scavenging and antitumour activities. The present investigation was designed to evaluate the anticarcinogenic potential Accepted of EIC against diethylnitrosamine (DEN) induced hepatocellular June 24, 2017 carcinoma in male Wistar rats. Hepatocarcinogenesis was induced by a single intraperitoneal administration of DEN (200 mg/kg) and Released the carcinogenic effect was promoted by phenobarbital given through drinking water for 16 successive weeks. EIC at the dose of June 30, 2017 200 and 400 mg/kg/day were administered orally for the entire study Open Acess period. After the end of experimental period, changes in body Full Text Article weight, the weight of liver, relative liver weight, lipid peroxidation, antioxidant, serum hepatic parameters, tumour markers, DNA, RNA and protein content were analysed. Treatment with EIC significantly increased the body weight (P < 0.01-0.001), reduced the liver weight and relative liver weight (P < 0.01-0.001), restored the altered serum hepatic parameters (P < 0.001), down-regulated the serum tumour markers such as alpha-fetoprotein and carcinoembryonic antigen (P < 0.001) when compared to DEN group. EIC treatment restored the antioxidant enzymes and significantly reduced the lipid peroxidation in DEN-treated animals (P < 0.001). EIC treatment also significantly reduced the elevated levels of nucleic acid levels and restored the protein content in liver tissues (P < 0.001). We ORCID investigated the anticarcinogenic potential of EIC against DEN- 0000-0003-1309-3886 induced HCC in rats. Chemoprotective effect of the extract might be Raju Senthil Kumar related with antioxidant, free radical scavenging and reduction of 0000-0001-6472-9173 lipid peroxidation. The results suggested that EIC would be a potent Gopal Murugananthan anticarcinogenic agent inhibiting DEN-induced hepatic carcinoma. 0000-0002-1125-7800 Keywords: Indigofera cordifolia; Diethylnitrosamine; Serum Balasubramanian hepatic parameters; Carcinoembryonic antigen; Alpha-fetoprotein; Rajkapoor Chemoprevention. Introduction around the globe. Increase in life expectation and adopting western diet and Cancer is a worldwide challenge lifestyles, tobacco use and exposure to and remains the largest reason of death carcinogens are the principal factors for ISSN 2358-2731/BJBS-2017-0019/4/7/7/53 Braz. J. Biol. Sci. http://revista.rebibio.net 54 Kumar et al. increasing the burden of cancer in the and prognosis of HCC, chemoprevention developing countries. Hepatocellular has considered being the best approach in carcinoma (HCC) is the second leading lowering the morbidity and death associated cause of cancer mortality throughout the with this disease. Chemoprevention is the world (Jemal et al., 2011). It has been modern strategy to manage primary liver joined with totally different etiologies such cancer. It is the mode of cancer as chronic hepatitis B and C infection and management during which the prevalence alcohol use (Shih et al., 2009; Kumar et al., of the disease are often prevented, delayed 2011). Because of high tolerance of liver, or reversed by the administration of one or HCC is never detected at the early phase more natural and/or synthetic agent. and treatment includes poor prognosis in Epidemiologic evidence have exposed the most of the cases, creating it a very chemopreventive effect of the many important health problem (El-Serag and medicinal plants and their phytochemicals Rudolph, 2007). The relapse rates of HCC in the prevention of the disease. are high and also the survival rate of the Indigofera cordifolia Roth. patient with HCC has not increased in the (Fabales: Fabaceae) it is an Indian past few decades. Surgery, including medicinal plant and has long been used by transplantation, is presently the foremost ethnic people and native medical successful treatment for this frightful practitioners to treat diseases like disease. rheumatism, arthritis, inflammation, The liver is the major organ that neoplasm and liver diseases. Literature involves in the metabolism of many survey revealed that crude methanol and xenobiotics; hence, it is more liable to aqueous extracts of seeds of this plant has carcinogenic insult. Diethylnitrosamine potent antioxidant, radical scavenging and (DEN) is a potent environment carcinogen wound healing activities (Tambe et al., that is entered through the food chain. DEN 2010; Sarin and Upman, 2011; Khatri et al., is synthesised endogenously and located in 2013). Earlier studies on this plant in our occupational settings, tobacco smoke, laboratory revealed that the ethanol extract processed meat, alcoholic beverages, of entire plant possesses potent antioxidant, agricultural chemicals, cosmetics and radical scavenging and antineoplastic pharmaceutical substances. It is made from properties in transplantable tumour models metabolism of certain medication. It causes (Thangavel et al., 2014; Thangavel et al., proliferative, chronic and cancerous lesions 2015). Based on the results obtained from in the liver. It will alkylate DNA with itself our previous works, an attempt was made to being converted to the extremely reactive check the anticarcinogenic impact of molecule by CYP450 dependent ethanol extract of I. cordifolia (EIC) against oxygenases and generates reactive oxygen DEN-induced phenobarbitone promoted species (ROS) inflicting oxidative stress. It hepatocarcinogenesis in rat model. also induces chromosomal aberrations, micronuclei and chromatid exchanges in the Materials and methods liver and these mutations evoked by DEN are responsible for the event of carcinoma Chemicals (Sivaramakrishnan et al., 2008; Mandal et Diethylnitrosamine (DEN) and al., 2008; Jagadeesh et al., 2009). The rat phenobarbitone were purchased from model of DEN-induced carcinoma is the Sigma Chemicals Co. (St. Louis, MO, foremost accepted and widely used USA). Carboxy methyl cellulose (CMC) experimental model to study the was procured from Loba Chemie (Mumbai, chemopreventive and chemotherapeutic India). All other chemicals used for the potential of many drugs. Human liver experiments were of analytical grade. metabolises DEN similar to that of rat liver with relevance such as genetic alteration, Animals morphology and gene expression. Male Wistar rats (150-160 g) were Considering the narrow treatment options purchased from Venkateshwara Enterprises, Braz. J. Biol. Sci., 2017, v. 4, No. 7, p. 53-66. Anticarcinogenic potential of Indigofera cordifolia 55 Bangalore, India. They were maintained in 400 mg/kg body weight (Thangavel et al., standard microlon boxes and were given 2015), by oral route for sixteen consecutive standard laboratory diet and water ad weeks on five days per week. At the end of libitum. The study was conducted after experiments, blood was collected from obtaining Institutional Animal Ethical overnight fasted animals by retro-orbital Committee (IAEC) clearance and the plexus method after anaesthetized slightly animals were handled as per the Committee with anaesthetic ether and serum was for the Purpose of Control and Supervision separated out. The animals were sacrificed of Experiments on Animals (CPCSEA) and the liver was immediately removed, guidelines of Good Laboratory Practice. washed with ice-cold saline and subsequently blotted with filter paper, then Plant material and extraction weighed. A small portion of liver was fixed The whole plant of I. cordifolia was in 10% formalin solution for collected from Tiruchengode, Namakkal histopathological studies. District, Tamilnadu, in the month of November 2015. The plant was then Serum biochemical and tumour authenticated by a botanist from Botanical markers estimation Survey of India, Coimbatore and a voucher Serum was analysed for aspartate specimen is preserved in our laboratory for aminotransferase (AST), alanine further reference (Voucher specimen No. P. aminotransferase (ALT), alkaline Chem/SVCP/15-16/IC-0213). The shade phosphatase (ALP), gamma glutamate dried, powdered plant material was transpeptidase (GGTP), total cholesterol extracted with 80% v/v ethanol by and total bilirubin by using commercial kits continuous hot percolation process using (Agappe diagnostics and Ecoline soxhlet apparatus for 72 h. The extract was Diagnostic Kits, India) following the filtered and concentrated to dryness under manufacturer’s instruction. Liver tumour reduced pressure and controlled markers like Alpha-fetoprotein (AFP) and temperature (50 °C-60 °C) in a rotary Carcinoembryonic antigen (CEA) in rat evaporator. A brownish yellow solid mass serum were estimated by using obtained was stored in a vacuum desiccator commercially available ELISA kits (USCN until further use. life science Inc. Wuhan, China) according to the manufacturer’s instruction. A 10% Experimental
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