Skip to main content
SpringerLink
Log in

Chemopreventive and antioxidant activity by Smilax zeylanica leaf extract against N-nitrosodiethylamine induced hepatocarcinogenesis in wistar albino rats

  • Research Article
  • Published:
Oriental Pharmacy and Experimental Medicine Aims and scope Submit manuscript

Abstract

The chemopreventive potential of methanol extract of Smilax zeylanica leaves against N-nitrosodiethylamine (NDEA) induced hepatocarcinogenesis was investigated in wistar albino male rats. The animals were divided into six groups. Group I served as normal control group, group II and group III served as extract control received Smilax zeylanica leaf extract 200 mg/kg and 400 mg/kg respectively once daily orally. A single intraperitoneal injection of NDEA (200 mg/kg) to group IV, group V and group VI animals followed by a single dose of carbon tetrachloride (CCl4, 2 ml/kg) 2 weeks later induced hepatocarcinogenesis in rats at the end of experimental period (16 weeks). Starting 1 week prior to NDEA administration, group V and group VI animals received methanol extract of Smilax zeylanica 200 mg/kg and 400 mg/kg respectively once daily orally for 16 weeks. At the end of the experimental period, the body weight, morphology of liver, relative liver weight, the levels of liver injury and liver cancer markers, the levels of lipid peroxidation and the activities of enzymatic and non-enzymatic antioxidants in liver tissue were assessed. Group IV NDEA induced hepatocarcinoma animals showed body weight loss with a significant increase in relative liver weight. The levels of liver injury and liver cancer markers such as alanine transaminase (AST), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), α-feto protein (AFT) and carcinoembryonic antigen (CEA) were substantially increased in group IV untreated animals. Moreover, with reference to lipid peroxidation and antioxidant system, group IV animals showed elevated levels of lipid peroxidation with a subsequent decrease in activities of antioxidant enzymes viz., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), glutathione-S-transferase (GST), glutathione (GSH), vitamin- C and vitamin-E in liver tissue. In contrast, the NDEA induced animals in group V and group VI treated with Smilax zeylanica leaf extract showed a significant improvement in body weight gain at the end of experimental period (16 weeks). A significant decrease in relative liver weight with reduced levels of liver injury and liver cancer markers in serum were observed in treatment groups. Moreover, treatment with Smilax zeylanica leaf extract decreased the extent of lipid peroxidation with concomitant increase in activities of enzymatic and non-enzymatic antioxidants when compared with NDEA induced untreated group IV animals. The histological observations of liver tissue too correlated the biochemical observations. Furthermore, the extract did not produce any deleterious effects in extract alone treated groups. These findings suggest that, the potential chemoprevention of methanol extract of Smilax zeylanica leaves might be due to stabilization and increase in all the components of antioxidant system attributed to antioxidant and free radical scavenging activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Abdala S, Martin-Herrera D, Benjumea D, Perez-Paz P (2008) Diuretic activity of Smilax canariensis, an endemic canary Islend species. J Ethnopharmacol 119:12–16

    Article  CAS  PubMed  Google Scholar 

  • Abelev GI (1971) Alpha-feto protein in association with malignant tumours. Adv Cancer Res 14:295–357

    Article  CAS  PubMed  Google Scholar 

  • Akintonwa DA (1985) The derivation of nitrosamines from some therapeutic amines in the human environment. Ecotoxicol Environ Saf 9:64–70

    Article  CAS  PubMed  Google Scholar 

  • Allen RG (1991) Oxygen-reactive species and antioxidant responses during development: the metabolic paradox of cellular differentiation. Proc Soc Exp Biol Med 196:117–129

    Article  CAS  PubMed  Google Scholar 

  • Ambasta SP (2006) The useful plants of India. NISCAIR, New Delhi, p 578

    Google Scholar 

  • Anita A, Purnima A, Madhavan V, Raju A (2010) In- vitro and in-vivo antioxidant activity studies on the leaves of Smilax zeylanica L. (Smilacaceae). J Pharm Res 3(10):2427–2430

    Google Scholar 

  • Bansal AK, Trivedi R, Soni GL, Bhatnagar D (2005) Protective role of vitamin E pre-treatment on N-nitrosodiethylamine induced oxidative stress in rat liver. Chem Biol Interact 156:101–111

    Article  CAS  PubMed  Google Scholar 

  • Barker H, Frank O, Angelis B, Feingold S (1951) Plasma tocopherol in man at various times after ingesting free or acetylated tocopherol. Nutr Rep Int 21:531–536

    Google Scholar 

  • Bebe FN, Panemangalore M (2003) Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissues of rats. J Environ Sci Health B38:349–363

    Article  CAS  Google Scholar 

  • Befeler AS, Di Bisceglie AM (2002) Hepatocellular carcinoma: diagnosis and treatment. Gasteroenterology 122(6):1609–1619

    Article  Google Scholar 

  • Bhattacharjee R, Sil PC (2006) The protein fraction of Phyllanthus niruri plays a protective role against acetaminophen induced hepatic disorder via its antioxidant properties. Phytother 20:595–601

    Article  Google Scholar 

  • Bhattracharya S, Chatterjee M (1998) Protective role of Trianthema portulacastrum against diethylnitrosamine- induced experimental hepatocarcinogenesis. Cancer Lett 129:7–13

    Article  Google Scholar 

  • Bhosale P, Motiwale L, Ignle AD, Gadre RV, Rao KVK (2002) Protective effect of Rhodotorula glutinis NCIM3353 on the development of hepatic preneoplastic lesions. Curr Sci 83(3):303–308

    Google Scholar 

  • Bishayee A, Dhir N (2009) Resveratrol-mediated chemoprevention of diethylnitrosamine-initiated hepatocarcinogenesis: inhibition of cell proliferation and induction of apoptosis. Chem Biol Interact 179(2–3):131–144

    Article  CAS  PubMed  Google Scholar 

  • Bulle F, Mavier P, Zafrani ES, Preaux AM, Lescs MC, Siegrist S, Dhumeaux D, Guellaen G (1990) Mechanism of gamma-glutamyl transpeptidase release in serum during intra hepatic and extra hepatic cholestasis in the rat: a histochemical, biochemical and molecular approach. Hepatology 11:545–550

    Article  CAS  PubMed  Google Scholar 

  • Chaudiere J (1994) Some chemical and biochemical constrains of oxidative stress in living cells. In: Rice-Evans CA, Burdon RH (eds) Free radical damage and its control. Elsevier Science, Amsterdam, pp 25–26

    Chapter  Google Scholar 

  • Das S (1994) Vitamin E in the genesis and prevention of cancer. A review. Acta Oncol 33:615–619

    Article  CAS  PubMed  Google Scholar 

  • David EJ (1999) Special considerations in interpreting liver function tests. Am Family Phy 59:8

    Google Scholar 

  • Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77

    Article  CAS  PubMed  Google Scholar 

  • Farombi EO, Olowg BI, Emerole GO (2000) Effect of three structurally related antimalarial drugs on liver microsomal components and lipid peroxidation in rats. Comp Biochem Physiol 126:217–224

    CAS  Google Scholar 

  • Feo F, Pascale RM, Simile MM, De Miglio MR, Muroni MR, Calvisi D (2000) Genetic alterations in liver carcinogensis; implications for new preventive and therapeutic strategies. Crit Rev Oncog 11:19–62

    Article  CAS  PubMed  Google Scholar 

  • Folch J, Lees M, Solane SGH (1957) A simple method for isolation and purification of total lipids from animal tissues. J Bio Chem 26:497–509

    Google Scholar 

  • Gamble JS (2004) Flora of the Presidency of Madras. Dehradun: Bishen Singh, Mahendra Pal Singh eds, pp. 1518

  • George J (2003) Ascorbic acid concentrations in diethylnitrosamine induced hepatic fibrosis in rats. Clin Chem Acta 335:39–47

    Article  CAS  Google Scholar 

  • Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    CAS  PubMed  Google Scholar 

  • Hai W, Kim C, Song S, Kang C (2001) Study on mechanism of multistep hepatotumorigenesis in rat: development of hepatotumorigenesis. J Vet Sci 2:53–58

    Google Scholar 

  • Halsted CH, Villanueva J, Chandler CJ, Stabler SP, Allen RH, Muskhelishvili L, James SJ, Poirier L (1996) Ethanol feeding of micropigs alters methionine Metabolism and increases hepatocellular apoptosis and proliferation. Hepatology 23(3):497–505

    Article  CAS  PubMed  Google Scholar 

  • Horn HD, Burns FH (1978) Assay of glutathione reductase activity. In: Bergmeyer HV (ed) Methods of enzymatic analysis. Academic, New York, pp 142–146

    Google Scholar 

  • IARC (1972) Monograph on the evaluation of carcinogenic risk of chemicals to man, vol 1. International Agency for Research on Cancer, Lyon, pp 107–124

    Google Scholar 

  • Jagadeeswaran R, Thirunavukkarasu C, Nalini R, Gunasekaran S, Sakthisekaran D (2000) In vitro studies on the selective cytotoxic effect of crocetin and quercetin. Fitotherapia 71(4):395–399

    Article  CAS  Google Scholar 

  • Jahan MS, Vani G, Shyamaladevi CS (2011) Anti-carcinogenic effect of Solanum trilobatum in diethylnitrosamine induced and Phenobarbital promoted hepatocarcinogenesis in rats. Asian J Biochem 6(1):74–81

    Google Scholar 

  • Karkar P, Das B, Viswanath PN (1984) Modified spectrophotometer assay of SOD. Ind J Biochem Biophys 95:51–58

    Google Scholar 

  • Katiyar SK, Agarwal R, Mukhtar H (1993) Protective effects of green tea polyphenols administration by oral intubation against chemical carcinogen induced forestomach and pulmonary neoplasia in A/J mice. Cancer Lett 73:167–172

    Article  CAS  PubMed  Google Scholar 

  • Khandelwal KR, Kokate CK (1995) Pratical pharmacognosy, 4th edn. Vallabh Prakashan, New Delhi, p 110

    Google Scholar 

  • Kim J, Park EJ (2002) Cytotoxic anticancer candidates from natural resources. Curr Med Chem Anti-cancer Agents 2:485–537

    Article  CAS  Google Scholar 

  • Kind PRN, King EJJ (1954) Estimation of plasma phosphatase by determination of hydrolysed phenol with antipyrine. J Clin Pathol 7:322–330

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kirtikar KR, Basu BD (1991) Indian Medicinal Plants. Dehra Dun: Bishen Singh, Mahendra Pal Singh, eds, pp. 2496

  • Kovalsky I, Schaff Lapis K, Jeney A (1996) Marker enzymes of rat chemical hepatocarcinogenesis in human liver tumours. Pathol Oncol Res 2:54–55

    Google Scholar 

  • Kweon S, Park KA, Choi H (2003) Chemopreventive effect of garlic powder diet in diethylnitrosamine-induced rat hepatocarcinogenesis. Life Sci 73:2515–2526

    Article  CAS  PubMed  Google Scholar 

  • Liao DJ, Blanck A, Eneroth P, Gustafsson JA, Hällström IP (2001) Diethylnitrosamine causes pituitary damage, disturbs hormone levels and reduces sexual dimorphism of certain liver functions in the rat. Environ Health Perspect 109:943–947

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Liu JG, Zhao HJ, Liu YJ, Wang XL (2006) Effect of selenium-enriched malt on hepatocarcinogenesis, paraneoplastic syndrome and the hormones regulating blood glucose in rats treated by diethylitrosamine. Life Sci 78(20):2315–2321

    Article  CAS  PubMed  Google Scholar 

  • Loeppky RN (1994) Nitrosamine and nitroso compound chemistry and biochemistry. In: ACS Symposium Series 553, American Chemical Society, Washington, DC, pp. 1–12

  • Macnab GM, Urbanowicz JM, Kew MC (1978) Carcinoembryonic antigen in hepatocellular cancer. Br J Cancer 38:51–54

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Malloy HT, Evelyn KA (1937) The determination of bilirubin with the photometric colorimeter. J Biol Chem 119:481–490

    CAS  Google Scholar 

  • Mann J (2002) Natural products in cancer chemotherapy: past, present and future. Nat Rev Cancer 2:143–148

    Article  CAS  PubMed  Google Scholar 

  • Murray RK, Granner DK, Mayes PA (2003) Rodwell VW. Harper’s Illustrated Biochemistry, 26th ed, The McGraw-Hill Companies Inc.

  • Nadkarni KM (1976) Indian Materica Medica. Bombay Popular Prakashan, Bombay, p 1145

    Google Scholar 

  • Nwanjo HU, Oze GO (2007) Oxidative imbalance and non-enzymic antioxidant status in pulmonary tuberculosis infected. Pak J Nutr 6:590–592

    Article  Google Scholar 

  • OECD (2000) Guidance document on acute oral toxicity 425. Environmental health and safety monograph series on testing assessment. NO. 24

  • Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358

    Article  CAS  PubMed  Google Scholar 

  • Omaye ST, Turnbull JD, Sauberlich HE (1979) Selected methods for the determination of ascorbic acid in animal cells, tissues and fluids. Methods Enzymol 62:1–11

    Google Scholar 

  • Oommachan, Masih SK (1991) Ethnobotanical and conservational aspects of medicinal plants of Madhya Pradesh. Indian J Pure Appl Biol 6(1):39–44

    Google Scholar 

  • Parola M, Robino G (2001) Oxidative stress-related molecules and liver fibrosis. J Hepatol 35:297–306

    Article  CAS  PubMed  Google Scholar 

  • Rajesh V, Perumal P, Sundarrajan T (2010) Antidiabetic activity of methanol extract of Smilax zeylanica Linn in streptozotocin induced diabetic rats. Internet J Pharmacol 6(1)

  • Ramakrishnan G, Ragahavendran HR, Vinodkumar R, Devaki T (2006) Suppression of N-nitrosodiethylamine induced hepatocarcinogenesis by silymarin in rats. Chem Biol Interact 161:104–114

    Article  CAS  PubMed  Google Scholar 

  • Ramakrishnan G, Augustine TA, Jagan S, Vinodhkumar R, Devaki T (2007) Effect of silymarin on N-nitrosodiethylamine induced hepatocarcinogenesis in rats. Exp Oncol 29(1):39–44

    CAS  PubMed  Google Scholar 

  • Ramaswamy SN, Radhakrishna Rao M, Govindappa (2001) Flora of Shimoga district Karnataka. Prasaranga, Mysore, p 619

    Google Scholar 

  • Reitman S, Frankel SA (1957) Coloirmetric method for determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases. Am J Clin Pathol 28:56–63

    CAS  PubMed  Google Scholar 

  • Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973) Selenium: biochemical role as a component of glutathione peroxidase. Science 179:588–590

    Article  CAS  PubMed  Google Scholar 

  • Saldhana CJ, Wicolson DH (1976) Flora of Hassan district of Karnataka. Amerind Pub Co Pvt Ltd, New Delhi, p 804

    Google Scholar 

  • Sallie R, Tredger JM, Willam R (1991) Drug and the liver. Biopharm Drug Dispos 12:251–259

    Article  CAS  PubMed  Google Scholar 

  • Santapau H, Henry AN (1976) A dictionary of the flowering plants in India (reprint). CSIR, New Delhi, p 58

    Google Scholar 

  • Sell S, Beckar FF (1978) Alpha-feto protein. Natl Cancer Inst 60:19–26

    CAS  Google Scholar 

  • Shao B, Guo HZ, Cui YJ, Ye M, Han J, Guo DA (2007) Steroidal saponins from Smilax china and their anti-inflammatory activities. Phytochemistry 68:623–630

    Article  CAS  PubMed  Google Scholar 

  • Shijun L, Tao Y, Yang J-Q, Terry DO, Larry WO (2000) The role of cellular glutathione peroxidase redox regulation in the suppression of tumour cell growth by manganese superoxide dismutase. Cancer Res 60:3927–3939

    Google Scholar 

  • Siegers CP and Younes M (1981). Effect of bioflavonoids on lipid peroxidation induced by glutathione depletion. Pro Int Bioflavonoid Symposium, Munich, FRG, pp. 409

  • Sikkim H, Mulee B (2000) Lipid peroxidation and antioxidant enzymes in the blood of rats treated with benzo[a]pyrene. Chem Biol Interact 127:139–150

    Article  Google Scholar 

  • Sinha AK (1972) Colorimetric assay of catalase. Anal Biochem 47:389–394

    Article  CAS  PubMed  Google Scholar 

  • Sivaramakrishnan V, Shilpa PNM, Kumar VRP, Devaraj SN (2008) Attenuation of N-nitrosodiethylamine induced hepatocellular carcinogenesis by a novel flavonol - Morin. Chem Biol Interact 171:79–88

    Article  CAS  PubMed  Google Scholar 

  • Soni KB, Lahiri M, Chakradeo P, Bhide SV, Kuttan R (1997) Protective effect of food additives on aflatoxin-induced mutagenicity and hepatocarcinogenicity. Cancer Lett 115:129–133

    Article  CAS  PubMed  Google Scholar 

  • Szas G (1976) Reaction rate method for gamma glutamyl transferase activity in serum. Clin Chem 22:2031–2055

    Google Scholar 

  • Takuji T, Toshihiro K, Toshihiko K, Aijin W, Masumi S, Kiyohisa O, Hideki M (1993) Inhibition of 4-nitroquinoline-1-oxide- induced rat tongue carcinogenesis by the naturally occurring plant phenolics caffeic, ellagic, chlorogenic and ferulic acids. Carcinogenesis 14(7):1321–1325

    Article  Google Scholar 

  • Thorgeirsson SS, Grisham JW (2002) Molecular pathogenesis of human hepatocellular Carcinoma. Nat Genet 31(4):339–346

    Article  CAS  PubMed  Google Scholar 

  • Verna L, Whysner J, Williams GM (1996) N-nitrodiethylamine mechanistic data and risk assessment: bioactivation, DNA-adduct formation, mutagenicity, and tumor intiation. Pharmacol Ther 71:57–81

    Article  CAS  PubMed  Google Scholar 

  • Wang XJ, Feng P, Wen ZY (1996) Study on invitro and invivo anticancer action of compound Smilax china L. Chin J Pathophysiol 12:614–1614

    Google Scholar 

  • Waris G, Ahsan H (2006) Reactive oxygen species: role in the development of cancer and various chronic conditions. J Carcinogen 5:14

    Article  Google Scholar 

  • Wattenberg LW, Estensen RD (1996) Chemopreventive effects of myo-inositol and dexamethasone on benzo[a]pyrene and 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone-induced pulmonary carcinogenesis in female A/J mice. Cancer Res 56:5132–5135

    CAS  PubMed  Google Scholar 

  • Wiseman H (1996) Dietary influences on membrane function: importance in protection against oxidative damage and disease. J Nutr Biochem 7:2–15

    Article  CAS  Google Scholar 

  • Wooten IDP (1964) Micro-analysis in medical biochemistry, 4th edn. J and A Churchill Ltd, London, pp 138–140

    Google Scholar 

  • Wroblewski F (1959) The clinical significance of transaminase activities of serum. Am J Med 27:911–923

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are thankful to the management of JKK Nattraja College of Pharmacy, Komarapalayam, Tamil Nadu, INDIA for providing necessary facilities to carry out the research work.

Author information

Authors and Affiliations

  1. Department of Pharmacology, JKK Nattraja College of Pharmacy, Natarajapuram, Komarapalayam, 638183, Namakkal District, Tamil Nadu, India

    Venugopalan Rajesh & Perumal Perumal

Authors
  1. Venugopalan Rajesh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Perumal Perumal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Venugopalan Rajesh.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rajesh, V., Perumal, P. Chemopreventive and antioxidant activity by Smilax zeylanica leaf extract against N-nitrosodiethylamine induced hepatocarcinogenesis in wistar albino rats. Orient Pharm Exp Med 14, 111–126 (2014). https://doi.org/10.1007/s13596-013-0125-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13596-013-0125-3

Keywords

Search

Navigation