Mast cells and pro-inflammatory cytokines roles in assessment of grape seeds extract anti-inflammatory activity in rat model of carrageenan-induced paw edema

Document Type: Short Communication


1 Department of Clinical Pathology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt

2 lecturer of clinical pathology,faculty of veterinary medicine,zagazig university

3 MVSc of Clinical Pathology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt


Objective(s): Reactive oxygen species (ROS)-produced oxidative disorders were involved at the pathophysiology of many inflammatory processes via the generation of pro-inflammatory cytokines and antioxidant defense system suppression. Although herbal antioxidants as mono-therapy relief many inflammatory diseases including, autoimmunity rheumatoid arthritis, but as combination therapy with other proven anti-inflammatory drugs in order to decreasing their toxic impacts has not yet been studied clearly, especially against chemical substances that’s induced local inflammation with characteristic edema.
Materials and Methods: Grape seeds extract (GSE) at a concentration of 40 mg/kg B. wt alone or in combination with indomethacin (Indo.) at a dose of 5 mg/Kg B. wt orally given for 10 days prior (gps VI, VII, VIII) or as a single dose after edema induction (gps IX, X, XI) in rat's left hind paw by sub-planter single injection of 0.1 carrageenan: saline solution (1%) (gp. V) to assess the prophylactic and therapeutic anti-inflammatory activities of both through  the estimation of selective inflammatory mediators and oxidative damage-related biomarkers as well as tissue mast cell scoring. Furthermore, both substances were given alone (gps II, III, IV) for their  blood, liver and kidney safety evaluation comparing with negative control rats (gp. I) which kept without medication.
Results: A marked reduction on the inflammatory mediators, edema volume and oxidative byproducts in edema bearing rats' prophylactic and treated with grape seeds extract and indomethacin was observed. Indomethacin found to induce some toxicological impacts which minimized when administered together with GSE.
Conclusion: GSE is a safe antioxidant agent with anti-inflammatory property.


Main Subjects

1. Mueller M, Hobiger S, Jungbauer A. Anti-inflammatory activity of extracts from fruits, herbs and spices. Food Chemist  J 2010; 122: 987–996.

2. Calixto JB, Medeiros R, Fernandes ES, Ferreira J, Cabrini DA, Campos  MM. Kinin B1 receptors: key G-protein-coupled receptors and their role in inflammatory and painful processes. British J Pharmacol 2004; 143:803-818.

3. Chakraborty  P, Das SBNN. A review: Some rhizomatous plants for the treatment of inflammation in West Bengal. World J Pharm and Pharmaceut  Sci 2016; 5: 889-897.

4. Inger  LM, Mart AFJ, van de L, Harald  EV. Non-steroidal anti-Inflammatory drugs: An overview of cardiovascular risks. Pharmaceut  (Basel) 2010; 3: 2146–2162.

5. Gian MA, Paola S, Giada A, Alessandro M, Luca L. Experimentally Induced Pulpal Lesion and Substance P Expression: Effect of Ketoprofen—A Preliminary Study. Internat J Dentis 2016; 2016: 1-5.

6. Abatan  MO, Lateef I, Taiwo VO. Toxic effects of non-steroidal anti-inflammatory agents in rats. Afric  J Biomed  Res 2006; 9:219–223.

7. Taiwo VO, Conteh OL. The rodenticidal effect of indomethacin: Pathogenesis and pathology. Veterin  Arch 2008;78:167-178.

8. Akpamu U,  Otamere HO, Ernest-Nwoke, IO, Ekhator CN, Osifo UC. The protective effect of testosterone on indomethacin induced gastric ulcer in female Sprague Dawley rats. Advan  in Endocrinol  2016; 2016: 1-5.

9. Malathi M, Sudarshana MS, Niranjan  MH. Anti-inflammatory effect of chloroform extract of Flaveria trinervia (Sprengel) C Mora –A medicinal herb. Internat  J  Pharmacol Biolog Sci 2012;4: 218- 221.

10. Schmitz  HH, Romanczyk L. Chocolate flavonoids: In vitro evidence suggesting a cardiovascular health benefit. (AAAS) Annu   Meet  and Sci  Innov Expos Washing 2000;DC. 17–22.

11. Schewe T, Kuhn  H, Sies H. Flavonoids of Cocoa inhibit recombinant human 5-lipoxygenase. J Nat  Med 2001a; 132: 1825–1829.

12. Schewe T, Sadik  C, Klotz  LO, Yoshimoto  T, Kuhn  H,  Sies  H, Polyphneols of cocoa: Inhibition of mammalian 15-lipoxygenase. J Biolog  Chemis 2001b; 382: 1687–1696.

13. Madkour N K, Ahmed M.  Amelioration of amiodarone-induced lung fibrosis in rats by grape seed extract. J Appl Sci Res 2013; 6:3698-3707.

14. Hanaa HA, Samiha MA, Fatehya MM, Nermeen MF. Biological significance of grape seed extract against lung injury induced by formaldehyde inhalation in rats. Der Pharma Chemica 2015; 7:64-78.

15. Romana  PA, Ceausu  RA, Cimpean AM, Serban T, Raica M, Gaje PN. Mast cells as key players in periodontal disease. Archiv  Biolog Sci 2014;66:801–809.

16. Wen-Guang L, Xiao-Yu Z, Yong-Jie W, Xuan T. Anti-inflammatory effect and mechanism of proanthocyanidins from grape seeds. Acta Pharmacol Sinica 200; 22:1117–1120.

17. Sandra H, Francesca C, Barbara C, Christopher JF. Inhibitors of fatty acid amide hydrolase reduce carrageenan-induced hind paw inflammation in pentobarbital-treated mice: comparison with indomethacin and possible involvement of cannabinoid receptors. Brit J Pharmacol 2005;  146:467–476.

18. Kakoti BB, Pradhan P, Borah S, Mahato K, Kumar M.    Analgesic and anti-Inflammatory activities of the methanolic stem bark extract of Nyctanthes arbor-tristis Linn. J  BioMed  Res  Internat 2013; 6: 826–830.

19. Brouckaert  P, Libert  C, Everaerdt B, Fiers W. Selective  species  specificity  of  tumor  necrosis  factor  for  toxicity  in  the  mouse.  Lymphok  Cytok Rex 1992; 193.

20. Grassi J, Roberge CJ, Frobert  Y. Determination of IL-1α,   IL-1β and IL-2 in biological media using specific enzyme immunometric assay. Immunol Res 1991; 119:125-145.

21. Wong  GG, Clark  SC. Multiple actions of interleukin 6 within a cytokine network. Immunol Today1988; 9:137-139.

22. Tatiya AU, Saluja AK. Further studies on membrane stabilizing, anti-inflammatory and FCA induced arthritic activity of various fractions of bark of Machilus macrantha in rats. Revista Brasileira de Farmacog 2011; 21: 1052-1064.

23. Manohar S, Sivanandhan  K, Michael A, Selvakumaran  R. Studies on asthma and allergic rhinitis among occupational hazardous workers in and around Pollachi. Arch Applied Sci Res J 2012; 4: 2237-2243.

24. Kosersky D, Brown J, Malone M.  Effects of cryogenine on adjuvant-induced arthritis and serum turbidity in rats. J Pharmaceut  Sci 1973; 62:1965–1968.

25. Uzkeser H, Cadirci E, Halici Z, Odabasoglu F, Polat  B,  Yuksel  T N, Zaltin S,  Atalay  F. Anti-Inflammatory and antinociceptive effects of salbutamol on acute and chronic models of inflammation in rats: Involvement of an antioxidant mechanism. Mediators of Inflamm  J 2012;167:382-390.

26. Koller A. Total serum protein. in: Kaplan, LA, Pesce, AJ, eds 1984; 1316-131.

27. Beutler E, Duron O, Kelly BM, Improved method for the determination of blood glutathione. J  Lab  Clin  Med 1963; 61: 882-888.

28. Nishikimi  M, Appaji  N, Yagi K. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem  Biophys  Res  Commun 1972; 46: 849-854.

29. Mihara M, Uchiyama M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal. Biochem 1978; 86: 271-278. 

30. Reitman S, Frankel S. Colorimetric method for determination of serum transaminase activity. Americ  J  Clin Pathol 1957; 28:56- 68.

31. Henry T J. Clinical chemistry: Principles and techniques. 2nd Ed., Brit  J  Hematol 1974; 13:68–70.

32. Doumas BT, Bayso DD, Carter RJ, Peters T, Schaffer  R. Determination of serum albumin. Clinica Chimica Act 1981; 27:1642.

33. Fawcett  JK , Scott JE. A rapid and precise method for the determination of urea. J Clin  Pathol 1960; 13:156.

34. Kind  PRN, King  EG. Estimation of plasma phosphatase by determination of hydrolyzed phenol with amino antipyrine. J Clin  Pathol 1954; 7:322-326.

35. Lillie  RD. Fixation In: Histopathologic technic and practical histochemistry. 3rd Ed., New York: McGraw Hill,1965; 32–60.

36. Bancroft JD, Gamble M. Theory and practice of histological techniques.4th Ed., Churchill, Livingston,2008.

37. SPSS  Statistical package for social science, computer software, Ver 16. London, UK: SPSS Company, 2008.

38. Sosa  S, Balick  M, Arvigo  R, Esposito  R, Pizza  C,  Altinier G. A screening of the topical anti-inflammatory activity of some central american plants. J Ethanopharmac 2002;  8:211-215.

39. Boakye-Gyasi E, Woode  E, Ainooson GK, Obiri DD, Ansah  C, Duwejua M, Donkoh A. Anti-Inflammatory and antipyretic effects of an ethanolic extract of Palisota hirsuta K. Schumroots. Afric  J Pharm  Pharmacol 2008; 2:191–199.

40. Angaji  SA, Mousavi SF, Babapour E. RNAa: A few key points. Ann  Biolog Res 2012; 3:3968-3977.

41. Terra X, Montagut G, Bustos M, Llopiz N, Ardèvol A, Bladé C, Fernández-Larrea  J, Pujadas  G, Salvadó  J, Arola  L, Blay M. Grape-seed procyanidins prevent low-GSPde inflammation by modulating cytokine expression in rats fed a high-fat diet. J Naut  and Biochem 2009; 20:210–218.

42. Abbas AM, Sakr HF. Effect of selenium and grape seed extract on indomethacin-induced gastric ulcers in rats. J Physiol  Biochem 2013;69: 527-537.

43. Li  W G, Zhang  XY, Wu YJ, Tian X. Anti-inflammatory effect and mechanism of proanthocyanidins from grape seeds. Act  Pharmacolo  Sinica 2001; 22:1117–1120. 

44. Young-Hua DH, Feng RZ, Li Q, Wei Q, Yin ZQ, Zhou L J, Tao  C, Jia RY. Anti-inflammatory activity of leaf essential oil from Cinnamomum longepaniculatum (Gamble) N. Chao. Intern J Clin Experim Med 2014;7:12-20.

45. Kumar S, Boehm J, Lee JC. MAP kinases: Key signalling molecules as therapeutic targets for inflammatory diseases. Nat Rev   Drug Discov 2003; 2:717–726.

46. Nishanthini D, Jesudoss PAC, Aravinda KB, Isabella T. Anti-inflammatory effect of aqueous extract of Emblica officinalis in animal models. Intern J Advan Res 2015; 3:671-677.

47. Ho SC, Hwang LS, Shen YJ, Lin CC. Suppressive effect of a proanthocyanidin-rich extract from longan (Dimocarpus longan Lour.) flowers on nitric oxide production in LPS-stimulated macrophage cells. J Agricult  Food Chemis 2007; 226:10664-10670.

48. Mohammad TM, Ali AH, Bahareh N, Seyyed  AM, Anahita R, Behnam G. A study of the mechanisms underlying the anti-inflammatory effect of ellagic acid in carrageenan-induced paw edema in rats. Ind J Pharmacol 2015; 3:292-298.

49. Lu XL, Feng-Lai Y, Fei-Hu C. Acid-sensing ion channels 3: a potential therapeutic target for pain treatment in arthritis.  Molecul  Biol  Report 2009; 37:3233-3238.

50. Abbas SS, Schaalan MF, Bahgat AK, El-Denshary ES. Possible potentiation by certain antioxidants of the anti-inflammatory effects of diclofenac in rats. Scient World J2014; 2:1-9. 

51. Lopez-Posadas  R, Ballester  I, Abadia-Molina  AC, Suarez MD, Zarzuelo A, Martinez- Augustin O, Sanchez MF. Effect of flavonoids on rat splenocytes, a structure-activity relationship study. Biochem Pharmacol 2008;76:495–506.

52. Saba  ZH, Kamaruddin MY, Suzana  M, Yasmin A, Mohd  Y. Gelam honey inhibits the production of pro-inflammatory, mediators NO, PGE2, TNF-α, and IL-6 in carrageenan-induced acute paw edema in rats. Alternat  Med  J 2012;10:96-136.

53. Piliero SJ, Colombo  C. Action of anti-inflammatory drugs on the lysozyme activity and “turbidity” of serum from rats with adjuvant arthritis or endocrine deficiency. J Pharmacol Exper Therap 1969; 165:294–299.

54. Weissmann  G. The role of lysosomes in inflammation and disease. Annu Rev Med 1967; 18:97-112.

55. Ziskoven  C, Jäger M, Kircher J, Patzer T, Bloch W, Brixius K, Krauspe R. Physiology and pathophysiology of nitrosative and oxidative stress in osteoarthritic joint destruction. Canad J Physiol Pharmacol 2011; 89:455-466.

56. Ho  E, Karimi  GK, Liu CC, Bhindi  R, Figtree GA.  Biological markers of oxidative stress: Applications to cardiovascular research and practice. Red  Biol 2013;1:483-491.

57. Saeidnia S, Abdollahi M. Toxicological and pharmacological concerns on oxidative stress and related diseases. Toxicol Appl Pharmacol 2013; 273:442–455.

58. Mohsin N, Mourad G, Faure M, Szawarc I, Bringer J. Metabolic syndrome performs better than the individual factors in predicting renal graft outcome. Transplant Proceed  J 2013;  45:3517-3519.

59. Bishayee A, Chatterjee M. Hepatoprotective activity of carrot (Daucus carota L.) against carbon tetrachloride intoxication in mouse liver. J Ethnopharmacol 1995;7:69-74.

60. Mariappan V, Amutha D, Shanthi S. Anti-inflammatory effect of Tarenna Asiatica (L) in carrageenan inducing inflammation. International J Pharm Pharmaceut Sci 2012;4:344-347.

61. Olaleye  SB, Farombi  EO. Attenuation of indomethacin- and HCl/ethanol-induced oxidative gastric mucosa damage in rats by kolaviron, a natural biflavonoid of Garcinia kola seed. Phytother Res J 2006;20:14-20.

62. Sehirli O, Ozel Y, Dulundu E, Topaloglu U, Ercan  F. Grape seed extract treatment reduces hepatic ischemia-reperfusion injury in rats. Phytother Res J 2008; 22: 43-48.

63. Shan Y, Ye X, Xin H. Effect of the grape seed proanthocyanidin extract on the free radical and energy metabolism indicators during the movement. Scient  Res  Essay 2010: 5:148-153.

64. Somaia  AN, Ayah  MHG, Ezz ESE, Abeer AS, Mona GK, Kawkab  AA. Protective effect of grape seed extract and/or silymarin against thioacetamide-induced hepatic fibrosis in rats. Liver J 2015;4:178.

65. Mahaveer G, Vikas S, Shreesh O, Jagriti  B, Dharmveer  SA. Anti-Inflammatory effect of Emblica officinalis in rodent  models of acute and chronic inflammation: Involvement of possible mechanisms. Intern  J  Inflamm 2014;1-6.

66. da Silva  EZM, Maria CJ , Constance  O. Mast cell function: A  new vision of an old cell. J  Histochem  Cytochem 2014; 62: 698–738.

67. Garbacki N, Tits M, Angenot L, Damas J. Inhibitory effects of proanthocyanidins from Ribes nigrum leaves on carrageenan acute inflammatory reactions induced in rats. BMC Pharmacol. J 2004; 4:25.

68. Jothi  S, Vetriselvan S, Gayathiri S, Ishwin  S, Shereenjeet  G, Devi CH, Yaashini A. Comparative evaluation of anti-inflammatory activity of extract of Curcuma longa and standard drug in carrageenan induced paw edema model using albino Wister rats. Internat  J Biolog  Pharmaceut Res 2012;3:538-544.

69. Walter  HH. Nonsteroidal anti-Inflammatory drugs and the kidney.  Pharmaceut 2010; 3:2291-2321

70. Marta K, Zadurska M, Cessak G, Fiedor P. Analysis of effect of non-steroidal anti-inflammatory drugs on teeth and oral tissues during orthodontic treatment. Report based on literature review. Acta Pol  Pharm 2013; 70:573-577.

71. Silva  RM, Beserra AM, Calegari PI, Souza MC, Dos Santos  RA, Lima JC, Balogun SO. Gastroprotective and ulcer-healing mechanisms of ellagic acid in experimental rats. J Agricult  Food Chemist 2011; 59: 6957–6965.

72. Yanarates  O, Guven  A,  Sizlan  A,  Uysal  B, Akgul O, Atim A, Ozcan A, Korkmaz A, Kurt E. Ameliorative effects of  proanthocyanidin on renal ischemia/reperfusion injury. Ren Fail J 2008; 30:931–938.

73. Ahmed AAE, Fatani AJ. Protective effect of grape seeds proanthocyanidins against naphthalene-induced hepatotoxicity in rats. Saud. Pharmaceut  J 2007; 15:38-47.

74. Ahmad  F, Khan G H. Study of aging and hepatoprotective activity of Vitis vinifera L. seeds in albino rats. Asian Pacific J Tropic Biomed 2012; 2:1770-1774.

75. Adedapo AA, Aiyelotan O. Effect of chronic administration of indomethacin on hematological parameters in rats. African J  Biomed Res 2001; 4:159 – 160.

76. Said  SE, Ayman EE, Hosam-Eldin HO, Hamdi MY. Amelioration of indomethacin systemic toxicity by gum arabic administration in adult albino rats. Intern. J.Med. Plant.& Alternat. Med 2014; 2:032-046.

77. Yamakoshi J, Saito M, Kataoka S, Kikuchi M. Safety evaluation of proanthocyanidin-rich extract from grape seeds. Food  Chem Toxicol  J 2002; 40:599–607.

78. Çetin A, Kaynar L, Kocyigit I, Hacioglu SK, Saraymen R, Ozturk A, Sari I, Sagdic O. Role of grape seed extract on methotrexate induced oxidative stress in rat liver. Americ J Chin Med 2008; 36:861-872.

79.Weiner ML, Nuber D, Blakemore WR, Harriman JF, Cohen SM. A 90-day dietary study of kappa carrageenan with emphasis on the gastrointestinal tract. Food  Chem Toxicol   J 2006; 45: 98-106.

80. Chis  IC, Ungureanu MI, Marton A, Simedrea  R, Muresan  A, Postescu ID, Decea N. Antioxidant effects of a grape seed extract in a rat model of diabetes mellitus. Diab Vascul Dis Res 2009;6: 200-204.

81. Coles  EH. Veterinary Clinical Pathology, 4th Ed., Phil- adelphia, PA: W.B. Saunders Company.1986.