Archivum Immunologiae et Therapiae Experimentalis

, Volume 62, Issue 6, pp 511–520 | Cite as

Effect of Diethylcarbamazine Citrate and Setaria equina Excretory–Secretory Material on Rat Hepatocellular Carcinoma

  • Mahmoud Abdel-Latif
  • Thabet Sakran
  • Gamal El-Shahawi
  • Hoda El-Fayoumi
  • Al-Mahy El-Mallah
Original Article


Diethylcarbamazine citrate (DEC) has been known for its efficacy to eradicate bancroftian filariasis in Egypt and other countries in the world. One of the known effects was to decrease the level of circulating filarial antigen in the patient’s serum. The target of this study was to examine the effect of DEC, excretory–secretory (ES) material from the filarial parasite Setaria equina or a combination of both on the status of oxidative stress and pathogenesis of rat hepatocellular carcinoma (HCC) induced by diethylnitrosamine and 2-acetylaminofluorene. This could be tested in vitro using nitroblue tetrazolium reduction test for measuring the level of superoxide anion (O 2 •− ) released from rat peritoneal macrophages. For in vivo test, a single dose before induction of carcinogenesis or continually repeated doses with DEC, ES or DEC + ES was used. Exposure of macrophages to ES could lead to a significant decrease (p < 0.01) in O 2 •− release, while DEC (200 μM) could modulate such effect with significant increase (p < 0.05). Pathogenesis of liver cancer and treatment were evaluated using histological investigation, level of antioxidant and liver function enzymes. Repeated ES doses could increase the activity of antioxidant enzymes, especially the catalase enzyme and show a protective effect on liver architecture. DEC could modulate the later effects when combined with ES. No significant effect on the liver function enzymes after treatment was observed. Nuclear factor κB was found to be localized only in the cytoplasm after single and repeated treatments with ES. This study could indicate the effect of S. equina ES as antioxidant against rat HCC, while DEC could modulate such effect when combined with it.


Diethylcarbamazine citrate Setaria equina Excretory–secretory material Hepatocellular carcinoma 



This work was supported by the Advanced Science Research Institute (ASRI) of Beni-Suef University, Beni-Suef city, Egypt. The authors had no commercial interest in the study.


  1. Bahgat MM, Saad AH, El-Shahawi GA et al (2011) Cross-reaction of antigen preparations from adult and larval stages of the parasite Setaria equina with sera from infected humans with Wuchereria bancrofti. East Mediterr Health J 17:679–686PubMedGoogle Scholar
  2. Bhattacharya C, Singh RN, Misra S et al (1997) Diethylcarbamazine: effect on lysosomal enzymes and acetylcholine in Wuchereria bancrofti infection. Trop Med Int Health 2:686–690PubMedCrossRefGoogle Scholar
  3. Bradford MM (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  4. Chenthamarakshan V, Reddy MV, Harinath BC (1996) Detection of filarial antigen by inhibition enzyme linked immunosorbent assay using fractionated Brugia malayi microfilarial excretory secretory antigen. J Biosci 21:27–34CrossRefGoogle Scholar
  5. Cohen G, Dembiec D, Marcus J (1970) Measurement of catalase activity in tissue extracts. Anal Biochem 34:30–38PubMedCrossRefGoogle Scholar
  6. Coleman SU, Klei TR, French DD (1985) Prevalence of Setaria equina (Nematode: Onchocercidae) in southeastern Louisiana horses. J Parasitol 71:512–513PubMedCrossRefGoogle Scholar
  7. Devi CM, Raj RK (1994) Immune response in mice against hatching associated materials from the filarial parasite Setaria digitata (Von Linstow). Indian J Exp Biol 32:848–853PubMedGoogle Scholar
  8. El-Shahawi GA, Abdel-Latif M, Saad AH et al (2010) Setaria equina: in vivo effect of diethylcarbamazine citrate on microfilariae in albino rats. Exp Parasitol 126:603–610PubMedCrossRefGoogle Scholar
  9. Espinoza E, Pérez-Arellano JL, Vicente B et al (2002) Cytoplasmic signalling pathways in alveolar macrophages involved in the production of nitric oxide after stimulation with excretory/secretory antigens of Toxocara canis. Parasite Immunol 24:535–544PubMedCrossRefGoogle Scholar
  10. Etienne A, Soulard C, Touvay C et al (1985) In vivo anti-allergic and anti-inflammatory effects of drugs that inhibit lipoxygenase activity in vitro. Int J Tissue React 7:459–462PubMedGoogle Scholar
  11. Farahnak A, Golestani A, Eshraghian M (2013) Activity of superoxide dismutase (SOD) enzyme in the excretory–secretory products of Fasciola hepatica and F. gigantica parasites. Iran J Parasitol 8:167–170PubMedCentralPubMedGoogle Scholar
  12. Gayral P, Gueyouche C, Bories C et al (1989) Macrofilaricidal activity of metabolites of diethylcarbamazine. Arzneimittelforschung 39:226–230PubMedGoogle Scholar
  13. González de Mejía E, Ramírez-Mares MV, Arce-Popoca E et al (2004) Inhibition of liver carcinogenesis in Wistar rats by consumption of an aqueous extract from leaves of Ardisia compressa. Food Chem Toxicol 42:509–516PubMedCrossRefGoogle Scholar
  14. Gonzàlez R, Ancheta O, Màrquez M et al (1994) Hepatoprotective effects of diethylcarbamazine in acute liver damage induced by carbon tetrachloride in rats. Zhongguo Yao Li Xue Bao 15:495–497PubMedGoogle Scholar
  15. Guo SP, Wang WL, Zhai YQ et al (2001) Expression of nuclear factor-κB in hepatocellular carcinoma and its relation with the X protein of hepatitis B virus. World J Gastroenterol 7:340–344PubMedGoogle Scholar
  16. Helming L, Böse J, Ehrchen J et al (2005) 1alpha, 25-dihydroxyvitamin D3 is a potent suppressor of interferon gamma-mediated macrophage activation. Blood 106:4351–4358PubMedCrossRefGoogle Scholar
  17. Helmy H, Weil GJ, Ellethy AS et al (2006) Bancroftian filariasis: effect of repeated treatment with diethylcarbamazine and albendazole on microfilaraemia, antigenaemia and antifilarial antibodies. Trans R Soc Trop Med Hyg 100:656–662PubMedCrossRefGoogle Scholar
  18. Hisadome M, Fukuda T, Terasawa M (1990) Effect of cysteine ethylester hydrochloride (Cystanin) on host defense mechanisms (V): potentiation of nitroblue tetrazolium reduction and chemiluminescence in macrophages or leukocytes of mice or rats. Jpn J Pharmacol 53:57–66PubMedCrossRefGoogle Scholar
  19. Hsiao WW, Liao HS, Lin HH et al (2013) Biophysical analysis of astrocytes apoptosis triggered by larval E/S antigen from cerebral toxocarosis-causing pathogen Toxocara canis. Anal Sci 29:885–892PubMedCrossRefGoogle Scholar
  20. Ikegami T, Matsuzaki Y, Fukushima S et al (2005) Suppressive effect of ursodeoxycholic acid on type IIA phospholipase A2 expression in HepG2 cells. Hepatology 41:896–905PubMedCrossRefGoogle Scholar
  21. Ishibashi N, Prokopenko O, Reuhl KR et al (2002) Inflammatory response and glutathione peroxidase in a model of stroke. J Immunol 168:1926–1933PubMedCrossRefGoogle Scholar
  22. Kar M, Mishra D (1976) Catalase, peroxidase and polyphenoloxidase activities during rice leaf senescence. Plant Physiol 57:315–319PubMedCentralPubMedCrossRefGoogle Scholar
  23. Klaunig JE, Kamendulis LM (2004) The role of oxidative stress in carcinogenesis. Annu Rev Pharmacol Toxicol 44:239–267PubMedCrossRefGoogle Scholar
  24. Ko RC, Fan L, Lee DL et al (1994) Changes in host muscles induced by excretory/secretory products of larval Trichinella spiralis and Trichinella pseudospiralis. Parasitology 108(Pt 2):195–205PubMedCrossRefGoogle Scholar
  25. Lammie PJ, Hightower AW, Richards FO Jr (1992) Alterations in filarial antigen-specific immunologic reactivity following treatment with ivermectin and diethylcarbamazine. Am J Trop Med Hyg 46:292–295PubMedGoogle Scholar
  26. Leiro JM, Alvarez E, Arranz JA et al (2003) In vitro effects of mangiferin on superoxide concentrations and expression of the inducible nitric oxide synthase, tumour necrosis factor-alpha and transforming growth factor-beta genes. Biochem Pharmacol 65:1361–1371PubMedCrossRefGoogle Scholar
  27. Li CK, Chung YY, Ko RC (1999) The distribution of excretory/secretory antigens during the muscle phase of Trichinella spiralis and T. pseudospiralis infections. Parasitol Res 85:993–998PubMedCrossRefGoogle Scholar
  28. Maizels RM, Denham DA (1992) Diethylcarbamazine (DEC): immunopharmacological interactions of an anti-filarial drug. Parasitology 105(Suppl):S49–S60PubMedCrossRefGoogle Scholar
  29. Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–674PubMedCrossRefGoogle Scholar
  30. Marzok MA, Desouky AR (2009) Ocular infection of donkeys (Equus asinus) with Setaria equina. Trop Anim Health Prod 41:859–863PubMedCrossRefGoogle Scholar
  31. Mauriz JL, Linares P, Macias RI et al (2003) TNP-470 inhibits oxidative stress, nitric oxide production and nuclear factor kappa B activation in a rat model of hepatocellular carcinoma. Free Radic Res 37:841–848PubMedCrossRefGoogle Scholar
  32. Menna C, Olivieri F, Catalano A et al (2010) Lipoxygenase inhibitors for cancer prevention: promises and risks. Curr Pharm Des 16:725–733PubMedCrossRefGoogle Scholar
  33. Mitsui Y, Takamura N, Fujimaki Y et al (1996) Development of a competitive enzyme-linked immunosorbent assay for diethylcarbamazine. Trop Med Int Health 1:528–534PubMedCrossRefGoogle Scholar
  34. Mohan S, Pradeep CG, Abhilash Kumar R et al (2001) The adult-specific ubiquinone Q(8) functions as an antioxidant in the filarial parasite, Setaria digitata. Biochem Biophys Res Commun 288:949–953PubMedCrossRefGoogle Scholar
  35. Mustafa H, Srivastava N, Kaushal DC et al (1996) Analysis and potential of excretory–secretory antigens of Setaria cervi for immunodiagnosis of human filariasis. Indian J Exp Biol 34:508–512PubMedGoogle Scholar
  36. Muthian G, Pradeep CG, Sargapradeep K et al (2006) Setaria digitata secreted filarial lipids modulate IL-12 signaling through JAK-STAT pathway leading to the development of Th1 response. Exp Parasitol 114:193–203PubMedCrossRefGoogle Scholar
  37. Nielsen NO, Simonsen PE, Dalgaard P et al (2007) Effect of diethylcarbamazine on HIV load, CD4%, and CD4/CD8 ratio in HIV-infected adult Tanzanians with or without lymphatic filariasis: randomized double-blind and placebo-controlled cross-over trial. Am J Trop Med Hyg 77:507–513PubMedGoogle Scholar
  38. Novoselov SV, Calvisi DF, Labunskyy VM et al (2005) Selenoprotein deficiency and high levels of selenium compounds can effectively inhibit hepatocarcinogenesis in transgenic mice. Oncogene 24:8003–8011PubMedCrossRefGoogle Scholar
  39. Pal BK, Kulkarni S, Bhandari Y et al (2006) Lymphatic filariasis: possible pathophysiological nexus with oxidative stress. Trans R Soc Trop Med Hyg 100:650–655PubMedCrossRefGoogle Scholar
  40. Pedersen BK, Bygbjerg IC, Svenson M (1987) Increase in natural killer cell activity during diethylcarbamazine treatment of patients with filariasis. Acta Trop 44:353–355PubMedGoogle Scholar
  41. Preuss HG, Jarrell ST, Scheckenbach R et al (1998) Comparative effect chromium vanadium and Gymnema sylvestre on sugar-induced blood pressure elevation in SHR. J Am Coll Nutr 17:116–123PubMedCrossRefGoogle Scholar
  42. Prince PR, Madhumathi J, Anugraha G et al (2013) Tandem antioxidant enzymes confer synergistic protective responses in experimental filariasis. J Helminthol. doi: 10.1017/S0022149X13000333 PubMedGoogle Scholar
  43. Raju K, Jambulingam P, Sabesan S et al (2010) Lymphatic filariasis in India: epidemiology and control measures. J Postgrad Med 56:232–238PubMedCrossRefGoogle Scholar
  44. Santos Rocha SW, Silva BS, Gomes FO et al (2012) Effect of diethylcarbamazine on chronic hepatic inflammation induced by alcohol in C57BL/6 mice. Eur J Pharmacol 689:194–203PubMedCrossRefGoogle Scholar
  45. Seki S, Nakashima H, Nakashima M et al (2011) Antitumor immunity produced by the liver Kupffer cells, NK cells, NKT cells, and CD8 CD122 T cells. Clin Dev Immunol 2011:868345PubMedCentralPubMedCrossRefGoogle Scholar
  46. Selkirk ME, Smith VP, Thomas GR et al (1998) Resistance of filarial nematode parasites to oxidative stress. Int J Parasitol 28:1315–1332PubMedCrossRefGoogle Scholar
  47. Singh A, Rathaur S (2005) Identification and characterization of a selenium-dependent glutathione peroxidase in Setaria cervi. Biochem Biophys Res Commun 331:1069–1074PubMedCrossRefGoogle Scholar
  48. Sutton RH, Atwell RB, Boreham PF (1985) Liver changes, following diethylcarbamazine administration, in microfilaremic dogs infected with Dirofilaria immitis. Vet Pathol 22:177–183PubMedCrossRefGoogle Scholar
  49. Thilagavathy AH, Prabha B, Raj RK (1990) Excretory secretory antigens of filarial parasite Setaria digitata. Indian J Exp Biol 28:291–292PubMedGoogle Scholar
  50. Tunon MJ, Sanchez-Campos S, Gutierrez B et al (2003) Effects of FK506 and rapamycin on generation of reactive oxygen species, nitric oxide production and nuclear factor kappaB activation in rat hepatocytes. Biochem Pharmacol 66:439–445PubMedCrossRefGoogle Scholar

Copyright information

© L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014

Authors and Affiliations

  • Mahmoud Abdel-Latif
    • 1
  • Thabet Sakran
    • 1
  • Gamal El-Shahawi
    • 1
  • Hoda El-Fayoumi
    • 1
  • Al-Mahy El-Mallah
    • 1
  1. 1.Zoology Department, Faculty of ScienceBeni-Suef UniversityBeni-SuefEgypt

Personalised recommendations