Skip to main content
SpringerLink
Log in

Protective effect of D-002, a mixture of beeswax alcohols, against indomethacin-induced gastric ulcers and mechanism of action

  • Original Paper
  • Published:
Journal of Natural Medicines Aims and scope Submit manuscript

Abstract

D-002, a mixture of higher aliphatic beeswax alcohols, produces gastroprotective and antioxidant effects. To investigate the gastroprotective effect of D-002 against indomethacin-induced ulcers, oxidative variables and myeloperoxidase (MPO) activity in the rat gastric mucosa were examined. Rats were randomized into six groups: a negative vehicle control and five indomethacin (50 mg/kg) treated groups, comprising a positive control, three groups treated orally with D-002 (5, 25 and 100 mg/kg) and one group with omeprazole 20 mg/kg intraperitoneally (ip). The contents of malondialdehyde (MDA), protein carbonyl groups (PCG), hydroxyl radical generation and catalase (CAT), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD) and MPO enzyme activities in the rat gastric mucosa were assessed. Indomethacin increased the content of MDA and PCG, the generation of *OH radical and MPO enzyme activity, while it decreased the CAT, GSH-PX and SOD activities as compared to the negative controls. D-002 (5–100 mg/kg) significantly and dose-dependently reduced indomethacin-induced ulceration to 75 %. Also, D-002 decreased the content of MDA and PCG, the generation of hydroxyl radicals and MPO activity as compared to the positive controls. The highest dose of D-002 (100 mg/kg) increased significantly GSH-PX and SOD activities, while all doses used increased CAT activities. Omeprazole 20 mg/kg, the reference drug, reduced significantly the ulcers (93 %), MDA and PCG, the generation of hydroxyl radicals and MPO activity, and increased the CAT, GSH-PX and SOD activities. D-002 treatment produced gastroprotective effects against indomethacin-induced gastric ulceration, which can be related to the reduction of hydroxyl radical generation, lipid peroxidation, protein oxidation and MPO activity, and to the increase of the antioxidant enzymes activities in the rat gastric mucosa.

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

Similar content being viewed by others

References

  1. Ramakrishnan K, Salinas RC (2007) Peptic ulcer disease. Am Fam Physician 76:1005–1012

    PubMed  Google Scholar 

  2. Tasman-Jones C (1986) Pathogenesis of peptic ulcer disease and gastritis: importance of aggressive and cytoprotective factors. Scand J Gastroenterol Suppl 122:1–5

    Article  PubMed  CAS  Google Scholar 

  3. Slomiany BL, Slomiany A (1991) Role of mucus in gastric mucosal protection. J Physiol Pharmacol 42:147–161

    PubMed  CAS  Google Scholar 

  4. Holzer P (2000) Gastroduodenal mucosal defense. Curr Opin Gastroenterol 16:469–478

    Article  PubMed  CAS  Google Scholar 

  5. Allen A, Flemström G (2005) Gastroduodenal mucus bicarbonate barrier: protection against acid and pepsin. Am J Physiol Cell Physiol 288:C1–C19

    PubMed  CAS  Google Scholar 

  6. Laine L, Takeuchi K, Tarnawski A (2008) Gastric mucosal defense and cytoprotection: bench to bedside. Gastroenterology 135:41–60

    Article  PubMed  CAS  Google Scholar 

  7. Wallace JL (2001) Pathogenesis of NSAID-induced gastroduodenal mucosal injury. Best Pract Res Clin Gastroenterol 15:691–703

    Article  PubMed  CAS  Google Scholar 

  8. Whittle BJ (2003) Gastrointestinal effects of nonsteroidal anti-inflammatory drugs. Fundam Clin Pharmacol 17:301–313

    Article  PubMed  CAS  Google Scholar 

  9. Lamarque D (2004) Pathogenesis of gastroduodenal lesions induced by non-steroidal anti-inflammatory drugs. Gastroenterol Clin Biol 28:C18–C26

    Article  PubMed  Google Scholar 

  10. Wallace JL, Keenan CM, Granger DN (1990) Gastric ulceration induced by nonsteroidal anti-inflammatory drugs is a neutrophil-dependent process. Am J Physiol 259:G462–G467

    PubMed  CAS  Google Scholar 

  11. Takeuchi K, Okada M, Ebara S, Osano H (1990) Increased microvascular permeability and lesion formation during gastric hypermotility caused by indomethacin and 2-deoxy-d-glucose in the rat. J Clin Gastroenterol 12:S76–S84

    Article  PubMed  Google Scholar 

  12. Pohle T, Brzozowski T, Becker JC, Van Der Voort IR, Markmann A, Konturek SJ, Moniczewski A, Domschke W, Konturek JW (2001) Role of reactive oxygen metabolites in aspirin-induced gastric damage in humans: gastroprotection by vitamin C. Aliment Pharmacol Ther 15:677–687

    Article  PubMed  CAS  Google Scholar 

  13. Yajima H, Yamao J, Fukui H, Takakura Y (2007) Up-to-date information on gastric mucosal lesions from long-term NSAID therapy in orthopedic outpatients: a study using logistic regression analysis. J Orthop Sci 12:341–346

    Article  PubMed  CAS  Google Scholar 

  14. Dhikav V, Singh S, Pande S, Chawla A, Anand KS (2003) Non-steroidal drug-induced gastrointestinal toxicity: mechanisms and management. J Indian Acad Clin Med 4:315–322

    Google Scholar 

  15. Yoshikawa T, Naito Y, Kishi A, Tomii T, Kaneko T, Iinuma S, Ichikawa H, Yasuda M, Takahashi S, Kondo M (1993) Role of active oxygen, lipid peroxidation, and antioxidants in the pathogenesis of gastric mucosal injury induced by indomethacin in rats. Gut 34:732–737

    Article  PubMed  CAS  Google Scholar 

  16. Naito Y, Yoshikawa T (2006) Oxidative stress involvement and gene expression in indomethacin-induced gastropathy. Redox Rep 11:243–253

    Article  PubMed  CAS  Google Scholar 

  17. Ganguly K, Kundu P, Banerjee A, Reiter RJ, Swarnakar S (2006) Hydrogen peroxide-mediated downregulation of matrix metalloprotease-2 in indomethacin-induced acute gastric ulceration is blocked by melatonin and other antioxidants. Free Radic Biol Med 41:911–925

    Article  PubMed  CAS  Google Scholar 

  18. Mizoguchi H, Ogawa Y, Kanatsu K, Tanaka A, Kato S, Takeuchi K (2001) Protective effect of rebamipide on indomethacin-induced intestinal damage in rats. J Gastroenterol Hepatol 16:1112–1119

    Article  PubMed  CAS  Google Scholar 

  19. Erkin B, Dokmeci D, Altaner S, Turan FN (2006) Gastroprotective effect of l-carnitine on indomethacin-induced gastric mucosal injury in rats: a preliminary study. Folia Med (Plovdiv) 48:86–89

    Google Scholar 

  20. Banerjee D, Maity B, Bauri AK, Bandyopadhyay SK, Chattopadhyay S (2007) Gastroprotective properties of Myristica malabarica against indometacin-induced stomach ulceration: a mechanistic exploration. J Pharm Pharmacol 59:1555–1565

    Article  PubMed  Google Scholar 

  21. Karakus B, Odabasoglu F, Cakir A, Halici Z, Bayir Y, Halici M, Aslan A, Suleyman H (2009) The effects of methanol extract of Lobaria pulmonaria, a lichen species, on indometacin-induced gastric mucosal damage, oxidative stress and neutrophil infiltration. Phytother Res 23:635–639

    Article  PubMed  Google Scholar 

  22. Carbajal D, Molina V, Valdés S, Arruzazabala ML, Más R (1995) Anti-ulcer activity of higher primary alcohols of beeswax. J Pharm Pharmacol 47:731–733

    Article  PubMed  CAS  Google Scholar 

  23. Carbajal D, Molina V, Valdés S, Arruzazabala ML, Rodeiro I, Más R, Magraner J (1996) Possible cytoprotective mechanism in rats of D-002, an anti-ulcerogenic product isolated from beeswax. J Pharm Pharmacol 48:858–860

    Article  PubMed  CAS  Google Scholar 

  24. Carbajal D, Molina V, Noa M, Valdés S, Arruzazabala ML, Aguilar C, Más R (2000) Effect of D-002 on gastric mucus composition in ethanol-induced ulcer. Pharmacol Res 42:329–332

    Article  PubMed  CAS  Google Scholar 

  25. Molina V, Valdés S, Carbajal D, Arruzazabala ML, Menéndez R, Más R (2001) Antioxidant effect of D-002 on gastric mucosa of rats with experimentally induced injury. J Med Food 4:79–83

    Article  PubMed  CAS  Google Scholar 

  26. Hano O, Illnait J, Más R, Fernández L, Piñol F, Fernández JC (2001) Effects of D-002, a product isolated from beeswax, on duodenal ulcer: a double-blind, placebo-controlled study. Curr Ther Res 62:394–407

    Article  CAS  Google Scholar 

  27. Illnait J, Terry H, Más R, Fernández L, Carbajal D (2005) Effects of D-002, a product isolated from beeswax, on gastric symptoms of patients with osteoarthritis treated with piroxicam: a pilot study. J Med Food 8:63–68

    Article  PubMed  CAS  Google Scholar 

  28. Fernández L, Terry H, Quiñones AM, Díaz B, Hernández ML, Illnait J, Mas R (2008) Effects of Abexol® in middle-aged and older subjects: an open follow-up. Rev CENIC Cien Biol 39:3–8

    Google Scholar 

  29. Rodríguez I, Illnait J, Terry H, Mas R, Fernández L, Fernández JC, Gámez R, Mesa M, Mendoza S, Ruiz D, Cruz Y (2009) Effects of Abexol® (beeswax alcohols) on gastrointestinal symptoms in middle-aged and older subjects. Rev CENIC Cien Biol 40:147–154

    Google Scholar 

  30. Menéndez R, Amor AM, González RM, Jiménez S, Más R (2000) Inhibition of rat microsomal lipid peroxidation by the oral administration of D002. Brazil J Med Biol Res 33:85–90

    Article  Google Scholar 

  31. Pérez Y, González RM, Amor AM, Jiménez S, Menéndez R (2002) D-002 on antioxidant enzymes in liver and brain of rats. Rev CENIC Cien Biol 33:3–5

    Google Scholar 

  32. Mendoza S, Noa M, Pérez Y, Mas R (2007) Preventive effect of D-002, a mixture of long-chain alcohols from beeswax, on the liver damage induced with CCl4 in rats. J Med Food 10:379–383

    Article  PubMed  CAS  Google Scholar 

  33. Menéndez R, Más R, Amor AM, Pérez Y, González RM, Fernández JC, Molina V, Jiménez S (2001) Antioxidant effects of D-002 on the in vitro susceptibility of whole plasma in healthy volunteers. Arch Med Res 32:436–441

    Article  PubMed  Google Scholar 

  34. Menéndez R, Más R, Illnait J, Pérez J, Amor AM, Fernández JC, González RM (2001) Effects of D-002 on lipid peroxidation in older subjects. J Med Food 4:71–77

    Article  PubMed  Google Scholar 

  35. López E, Illnait J, Molina V, Oyárzabal A, Fernández L, Pérez Y, Mas R, Mesa M, Fernández J, Mendoza S, Gómez M, Jiménez S, Ruiz D (2008) Effects of D-002 (beeswax alcohols) on lipid peroxidation in middle-aged and older subjects. Lat Am J Pharm 27:695–703

    Google Scholar 

  36. Rodríguez I, Illnait J, Molina V, Oyarzábal A, Fernández L, Fernández J, Mesa M, Mas R, Mendoza S, Gomez R, Jimenez S, Ruiz D (2010) Comparative antioxidant effects of beeswax alcohols and grape seed extract in healthy persons. Lat Am J Pharm 29:255–262

    Google Scholar 

  37. Ravelo Y, Molina V, Carbajal D, Fernández L, Fernández JC, Arruzazabala ML, Más R (2011) Evaluation of anti-inflammatory and antinociceptive effects of D-002 (beeswax alcohols). J Nat Med 65:330–335

    Article  PubMed  CAS  Google Scholar 

  38. Biswas K, Bandyopadhyay U, Chattopadhyay I, Varadaraj A, Ali E, Banerjee RK (2003) A novel antioxidant and antiapoptotic role of omeprazole to block gastric ulcer through scavenging of hydroxyl radical. J Biol Chem 278:10993–11001

    Article  PubMed  CAS  Google Scholar 

  39. Markwell MA, Haas SM, Bieber LL, Tolbert NE (1978) A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87:206–210

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  41. Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Methods Enzymol 233:357–363

    Article  PubMed  CAS  Google Scholar 

  42. Gutteridge JM, Halliwell B (1988) The deoxyribose assay: an assay both for ‘free’ hydroxyl radical and for site-specific hydroxyl radical production. Biochem J 253:932–933

    PubMed  CAS  Google Scholar 

  43. Burdon RH, Knippenberg PH (eds) (1991) Laboratory techniques in biochemistry and molecular biology, vol 22. Techniques in free radical research, vol 3. Elsevier, Amsterdam, pp 50–55

  44. Paglia DE, Valentine WN (1987) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70:158–169

    Google Scholar 

  45. Aebi H (1974) Catalase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Chemie, Weinheim, pp 673–684

    Chapter  Google Scholar 

  46. Worthington Biochemical Corporation (1972) Worthington enzyme manual. Freehold, New Jersey, pp 43–45

    Google Scholar 

  47. Suleyman H, Albayrak A, Bilici M, Cadirci E, Halici Z (2010) Different mechanisms in formation and prevention of indomethacin-induced gastric ulcers. Inflammation 33(4):224–234

    Article  PubMed  CAS  Google Scholar 

  48. Bayir Y, Odabasoglu F, Cakir A, Aslan A, Suleyman H, Halici M, Kazaz C (2006) The inhibition of gastric mucosal lesion, oxidative stress and neutrophil-infiltration in rats by the lichen constituent diffractaic acid. Phytomedicine 13:584–590

    Article  PubMed  CAS  Google Scholar 

  49. Odabasoglu F, Halici Z, Cakir A, Halici M, Aygun H, Suleyman H, Cadirci E, Atalay F (2008) Beneficial effects of vegetable oils (corn, olive and sunflower oils) and alpha-tocopherol on anti-inflammatory and gastrointestinal profiles of indomethacin in rats. Eur J Pharmacol 591:300–306. doi:10.1016/j.ejphar.2008.06.075

    Article  PubMed  CAS  Google Scholar 

  50. Yeo M, Kwak MS, Kim DK, Chung IS, Moon BS, Song KS, Hahm K-B (2006) The novel acid pump antagonists for anti-secretory actions with their peculiar applications beyond acid suppression. J Clin Biochem Nutr 38:1–8

    Article  CAS  Google Scholar 

  51. Simon WA, Sturm E, Hartmann HJ, Weser U (2006) Hydroxyl radical scavenging reactivity of proton pump inhibitors. Biochem Pharmacol 71:1337–1341

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre of Natural Products, National Centre for Scientific Research (CNIC), Ave 25 and 158, Cubanacán, P.O. Box 6880, Havana, Cuba

    Yohani Pérez, Ambar Oyárzabal, Rosa Mas, Vivian Molina & Sonia Jiménez

Authors
  1. Yohani Pérez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ambar Oyárzabal
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rosa Mas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vivian Molina
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sonia Jiménez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yohani Pérez.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pérez, Y., Oyárzabal, A., Mas, R. et al. Protective effect of D-002, a mixture of beeswax alcohols, against indomethacin-induced gastric ulcers and mechanism of action. J Nat Med 67, 182–189 (2013). https://doi.org/10.1007/s11418-012-0670-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11418-012-0670-y

Keywords

Search

Navigation