Cissampelos capensis and Pleiocarpa pycantha protect against N-nitrosodiethylamine-induced hepatotoxicity

  • Solomon E. Owumi
  • Saran O Nwozor
  • C O Igboekwe
  • Olubunmi A. Akinwumi


Pleiocarpa pycantha (PP) and Cissampellos capensis(CC) showed potential phytoprotection in vitro. We then investigated this effects against N-diethylnitrosamine (DEN) induced hepatiotoxicity and renal function in rat (n=5), treated as follows for 60 days: (1) Control (2) DEN, (3) DEN+PP (4) DEN+CC (5) DEN+PP+CC (6) PP, (7) CC only and group (8) PP+CC. Rats were sacrificed and organs harvested for Biochemical and histological examination. DEN-induced hepatic tumors were counted and measured, liver sections processed for histopathology. Furthermore, markers of oxidative stress, hepatotoxicity and renal function including superoxide dismutase, aspartate transaminase, and creatinine were assessed in serum, in addition to α-fetoprotein and blood glucose levels. DEN treatment resulted in weight loss; elevated serum levels of hepatic transaminases, α-fetoprotein and induction of T2-4 (tumour count) in experimental animals. Tumour incidences were reduced to T1-2, T1-2 and T1 in the presence of PP, CC and PP+CC co-treatment respectively. Histological investigation revealed increased growth in hepatic stellate cells and early fibrosis in DEN only treated group, which were partly reversed by PP and CC. Significant increase in blood glucose level was observed in DEN treated animals compared to PP and CC treated groups. In addition, DEN treated group were low in cellular antioxidant activity relative to other groups, which was elevated in the presence of PP and CC. Our findings indicated that PP and CC co-treatment alleviated DEN-induced hepatic tumorigenesis in exposed rats, reinforcing earlier in vitro findings. PP and CC phytoprotective effect maybe relevant in protecting the liver against chemical carcinogens.



Abdel-Monein A, Ahmed, O.M., Fahim, H.I., Zaky, M.Y. Ameliorative Effects of Quercetin and Naringenin on Diethylnitrosamine/2-acetyl aminoflourene-Induced Nephrotoxicity in Male Wistar Rats. American Journal of Biochemistry. 2016;6(5):113 -21.
Afzal M, Kazmi I, Khan R, Rana P, Kumar V, Al-Abbasi FA, et al. Thiamine potentiates chemoprotective effects of ibuprofen in DEN induced hepatic cancer via alteration of oxidative stress and inflammatory mechanism. Arch Biochem Biophys. 2017;623-624:58-63.
Aggarwal BB, Prasad S, Reuter S, Kannappan R, Yadev VR, Park B, et al. Identification of novel anti-inflammatory agents from Ayurvedic medicine for prevention of chronic diseases: "reverse pharmacology" and "bedside to bench" approach. Curr Drug Targets. 2011;12(11):1595-653.
Al-Rejaie SS, Aleisa AM, Al-Yahya AA, Bakheet SA, Alsheikh A, Fatani AG, et al. Progression of diethylnitrosamine-induced hepatic carcinogenesis in carnitine-depleted rats. World J Gastroenterol. 2009;15(11):1373-80.
Ames BN. Dietary carcinogens and anticarcinogens. Oxygen radicals and degenerative diseases. Science. 1983;221(4617):1256-64.
Astrog PG, S.; Berges, R.; Suschetet, M. No evidence for an inhibitory effect of beta-carotene or of canthaxanthin on the initiation of liver preneoplastic foci by diethylnitrosamine in the rat. Nutr Cancer. 1996;25:27 - 34.
Barbosa-Filho JMC-C, E.V.L.; Gray, A.I. Alkaloids of the Menispermaceae. Cordell GA, editor. Illinois: Academic Press; 2000.
Bartsch H, Hietanen E, Malaveille C. Carcinogenic nitrosamines: free radical aspects of their action. Free Radic Biol Med. 1989;7(6):637-44.
Basu A, Nguyen A, Betts NM, Lyons TJ. Strawberry as a functional food: an evidence-based review. Critical reviews in food science and nutrition. 2014;54(6):790-806.
Beutler E, Blume KG, Kaplan JC, Lohr GW, Ramot B, Valentine WN. International Committee for Standardization in Haematology: recommended methods for red-cell enzyme analysis. Br J Haematol. 1977;35(2):331-40.
Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and antioxidant defense. World Allergy Organ J. 2012;5(1):9-19.
Bode AM, Dong Z. Signal transduction and molecular targets of selected flavonoids. Antioxid Redox Signal. 2013;19(2):163-80.
Bostrom CE, Gerde P, Hanberg A, Jernstrom B, Johansson C, Kyrklund T, et al. Cancer risk assessment, indicators, and guidelines for polycyclic aromatic hydrocarbons in the ambient air. Environ Health Perspect. 2002;110 Suppl 3:451-88.
Brown JL. N-nitrosamines. Occup Med. 1999;14:839 -48.
Burkhill HM. The useful plants of the West Tropical Africa. 2nd ed. United Kingdom: Royal Botanical Gardens, Kew; 1985.
Chen YJ, Myracle AD, Wallig MA, Jeffery EH. Dietary broccoli protects against fatty liver development but not against progression of liver cancer in mice pretreated with diethylnitrosamine. J Funct Foods. 2016;24:57-62.
Chhabra SK, Souliotis VL, Kyrtopoulos SA, Anderson LM. Nitrosamines, alcohol, and gastrointestinal tract cancer: recent epidemiology and experimentation. In Vivo. 1996;10(3):265-84.
Chung MY, Lim TG, Lee KW. Molecular mechanisms of chemopreventive phytochemicals against gastroenterological cancer development. World J Gastroenterol. 2013;19(7):984-93.
De Wet H, Van Wyk, B.E.. An ethnobotanical survey of southern African Menispermaceae. S Afr J of Botany. 2008;74:2-9.
De Wet H. VHFR, Van Wyk B.-E. Alkaloids of Antizoma miersiana (Menispermaceae). Biochem Syst Ecol,. 2005;33:799-807.
De-Freitas MR D-AJ, Da-Cunha EVL, Barbosa-Filho JM, Gray AI. Milonine, an 8,14-dihydromorphinandienone alkaloid from leaves of Cissampelos sympodialis. Phytochemistry. 1995;40(5):1553 -15555.
Devic S. Warburg Effect - a Consequence or the Cause of Carcinogenesis? J Cancer. 2016;7(7):817-22.
El-Shahat M, El-Abd S, Alkafafy M, El-Khatib G. Potential chemoprevention of diethylnitrosamine-induced hepatocarcinogenesis in rats: myrrh (Commiphora molmol) vs. turmeric (Curcuma longa). Acta Histochem. 2012;114(5):421-8.
Ferrari N, Tosetti F, De Flora S, Donatelli F, Sogno I, Noonan DM, et al. Diet-derived phytochemicals: from cancer chemoprevention to cardio-oncological prevention. Curr Drug Targets. 2011;12(13):1909-24.
Gorman AA, Dastoor, N.J., Manfred, H., von Philipsborn, W., Renner, U., Schmid, H. . Alkaloids. CXXXII. Constitution of two new type dimeric indole alkaloids, pycnanthine and pleiomutinine. . Helv Chim Acta. 1969;52:33-55.
Grimm C, Hofstetter G, Aust S, Mutz-Dehbalaie I, Bruch M, Heinze G, et al. Association of gamma-glutamyltransferase with severity of disease at diagnosis and prognosis of ovarian cancer. Br J Cancer. 2013;109(3):610-4.
Gupta RK, Patel AK, Shah N, Chaudhary AK, Jha UK, Yadav UC, et al. Oxidative stress and antioxidants in disease and cancer: a review. Asian Pac J Cancer Prev. 2014;15(11):4405-9.
He WZ, Guo GF, Yin CX, Jiang C, Wang F, Qiu HJ, et al. Gamma-glutamyl transpeptidase level is a novel adverse prognostic indicator in human metastatic colorectal cancer. Colorectal Dis. 2013;15(8):e443-52.
Heindryckx F, Colle I, Van Vlierberghe H. Experimental mouse models for hepatocellular carcinoma research. Int J Exp Pathol. 2009;90(4):367-86.
Heinemann G, Vogt W. The determination of bilirubin with a new enzymatic method (Dri-STAT bilirubin) using the Hitachi 704 selective analyzer. Journal of clinical chemistry and clinical biochemistry Zeitschrift fur klinische Chemie und klinische Biochemie. 1988;26(6):391-7.
J. B. Dictionary of Natural Products on CD-ROM. . Chapman Hall, Hampden Data Services, Ltd: CRC Press; 2006.
Jen KL YS. Hepatotoxic actions of dietary amines. Toxicology and Ecotoxicology News. 1994;1:82-6.
Keawpradub N, Houghton PJ, Eno-Amooquaye E, Burke PJ. Activity of extracts and alkaloids of thai Alstonia species against human lung cancer cell lines. Planta Med. 1997;63(2):97-101.
Kim WR, Flamm SL, Di Bisceglie AM, Bodenheimer HC, Public Policy Committee of the American Association for the Study of Liver D. Serum activity of alanine aminotransferase (ALT) as an indicator of health and disease. Hepatology. 2008;47(4):1363-70.
Komakech R, Kang Y, Lee JH, Omujal F. A Review of the Potential of Phytochemicals from Prunus africana (Hook f.) Kalkman Stem Bark for Chemoprevention and Chemotherapy of Prostate Cancer. Evid Based Complement Alternat Med. 2017;2017:3014019.
Krinsky NI. Effects of carotenoids on cells. Molecular aspects of medicine. 1993;14(3):241-6.
Kumagai A. [Gout--its pathogenesis]. Kango Gijutsu. 1971;17(9):31-7.
Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: an overview. ScientificWorldJournal. 2013;2013:162750.
Liebler DC. Antioxidant reactions of carotenoids. Ann N Y Acad Sci. 1993;691:20-31.
Linnewiel-Hermoni K, Khanin M, Danilenko M, Zango G, Amosi Y, Levy J, et al. The anti-cancer effects of carotenoids and other phytonutrients resides in their combined activity. Arch Biochem Biophys. 2015;572:28-35.
Lucier G, ; Hook, G.E.R. Dietary Mutagens. Special Issue, Environ Health Perspect. 1986:67 - 72.
Luo WP, Fang YJ, Lu MS, Zhong X, Chen YM, Zhang CX. High consumption of vegetable and fruit colour groups is inversely associated with the risk of colorectal cancer: a case-control study. Br J Nutr. 2015;113(7):1129-38.
Magee PN. Nitrosamines and human cancer: introduction and overview. Eur J Cancer Prev. 1996;5 Suppl 1:7-10.
Manson MM. Cancer prevention -- the potential for diet to modulate molecular signalling. Trends Mol Med. 2003;9(1):11-8.
Milner JA, McDonald SS, Anderson DE, Greenwald P. Molecular targets for nutrients involved with cancer prevention. Nutr Cancer. 2001;41(1-2):1-16.
Misra HP, Fridovich I. The univalent reduction of oxygen by reduced flavins and quinones. J Biol Chem. 1972;247(1):188-92.
Murakami A, Ohigashi H, Koshimizu K. Anti-tumor promotion with food phytochemicals: a strategy for cancer chemoprevention. Biosci Biotechnol Biochem. 1996;60(1):1-8.
Odin AP. Vitamins as antimutagens: advantages and some possible mechanisms of antimutagenic action. Mutat Res 1997;386(1):39 - 67.
Park SY, Ollberding NJ, Woolcott CG, Wilkens LR, Henderson BE, Kolonel LN. Fruit and vegetable intakes are associated with lower risk of bladder cancer among women in the Multiethnic Cohort Study. J Nutr. 2013;143(8):1283-92.
Parola M, Robino G. Oxidative stress-related molecules and liver fibrosis. J Hepatol. 2001;35(2):297-306.
Peterson K. 'Natural' cancer prevention trial halted. Science. 1996;271(5248):441.
Peto R, Doll R, Buckley JD, Sporn MB. Can dietary beta-carotene materially reduce human cancer rates? Nature. 1981;290(5803):201-8.
Pflaum T, Hausler T, Baumung C, Ackermann S, Kuballa T, Rehm J, et al. Carcinogenic compounds in alcoholic beverages: an update. Arch Toxicol. 2016.
Phaniendra A, Jestadi DB, Periyasamy L. Free radicals: properties, sources, targets, and their implication in various diseases. Indian J Clin Biochem. 2015;30(1):11-26.
Pitot HC, Dragan YP. The multistage nature of chemically induced hepatocarcinogenesis in the rat. Drug Metab Rev. 1994;26(1-2):209-20.
Pradeep K, Raj Mohan CV, Gobianand K, Karthikeyan S. Protective effect of Cassia fistula Linn. on diethylnitrosamine induced hepatocellular damage and oxidative stress in ethanol pretreated rats. Biol Res. 2010;43(1):113-25.
Rashid S, Ali N, Nafees S, Hasan SK, Sultana S. Amelioration of renal carcinogenesis by bee propolis: a chemo preventive approach. Toxicol Int. 2013;20(3):227-34.
Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol. 1957;28(1):56-63.
Ribeiro FA, Gomes de Moura CF, Aguiar O, Jr., de Oliveira F, Spadari RC, Oliveira NR, et al. The chemopreventive activity of apple against carcinogenesis: antioxidant activity and cell cycle control. Eur J Cancer Prev. 2014;23(5):477-80.
Rossi T, Gallo C, Bassani B, Canali S, Albini A, Bruno A. Drink your prevention: beverages with cancer preventive phytochemicals. Polskie Archiwum Medycyny Wewnetrznej. 2014;124(12):713-22.
Rotruck JT, Pope AL, Ganther HE, Hoekstra WG. Prevention of oxidative damage to rat erythrocytes by dietary selenium. J Nutr. 1972;102(5):689-96.
Rouhollahi E, Moghadamtousi SZ, Al-Henhena N, Kunasegaran T, Hasanpourghadi M, Looi CY, et al. The chemopreventive potential of Curcuma purpurascens rhizome in reducing azoxymethane-induced aberrant crypt foci in rats. Drug Des Devel Ther. 2015;9:3911-22.
Satyanarayana K. Chemical examination of Scoparia dulcis (Linn): Part I. . Journal of Indian Chemical Society. 1969;46:765 - 6.
Shu LHPF. Alkylation of cellular macromolecules and target specificity of carcinogenic nitrosodialkylamines: metabolic activation by cytochromes P450 2B1 and 2E1. Carcinogenesis. 1997;18(4):801 - 10.
Sreepriya M, Bali G. Chemopreventive effects of embelin and curcumin against N-nitrosodiethylamine/phenobarbital-induced hepatocarcinogenesis in Wistar rats. Fitoterapia. 2005;76(6):549-55.
Sullivan BP, Meyer TJ, Stershic MT, Keefer LK. Acceleration of N-nitrosation reactions by electrophiles. IARC Sci Publ. 1991(105):370-4.
Surh YJ, Lee RC, Park KK, Mayne ST, Liem A, Miller JA. Chemoprotective effects of capsaicin and diallyl sulfide against mutagenesis or tumorigenesis by vinyl carbamate and N-nitrosodimethylamine. Carcinogenesis. 1995;16(10):2467-71.
Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nature reviews Cancer. 2003;3(10):768-80.
Trendowski M. Recent Advances in the Development of Antineoplastic Agents Derived from Natural Products. Drugs. 2015;75(17):1993 - 2016.
Van Wyk B.-E. GN. People's Plants: A guide to useful plants of southern Africa. Pretoria: Briza Publications; 2000.
Verna L, Whysner J, Williams GM. N-nitrosodiethylamine mechanistic data and risk assessment: bioactivation, DNA-adduct formation, mutagenicity, and tumor initiation. Pharmacology & therapeutics. 1996;71(1-2):57-81.
Watt J.M. B-BMG. The Medicinal and Poisonous Plants of Southern and Eastern Africa. 2nd ed. London: Livingstone; 1962.
Wu J, ; Lai, C,; Tsai, M,; Ho, C,; Wang, Y. Chemopreventive effect of natural dietary compounds on xenobiotic-induced toxicity. Journal of Food and Drug Analysis. 2016;25(1):176 - 86.
Xu M, Zhao Q, Shao D, Liu H, Qi J, Qin C. Chenodeoxycholic Acid Derivative HS-1200 Inhibits Hepatocarcinogenesis and Improves Liver Function in Diethylnitrosamine-Exposed Rats by Downregulating MTH1. Biomed Res Int. 2017;2017:1465912.
Yu MW ZY, Blaner WS, Santella RM. Influence of vitamins A, C, and E and beta-carotene on aflatoxin B1 binding to DNA in woodchuck hepatocytes. 1994;73(3):596 - 604.
Carcinogenesis/Oncology/Chemoprevention/Toxicology/Biomarkers/Oxidative Stress