The effects of low dose capsaicin on substance P expression in rat duodenum

Abstract

The aim of the present study was to investigate effects of low dose capsaicin (CAP) on substance P (SP) expression and histomorphometric properties of duodenum. In this study, 21-day-old rats were divided into three groups as control, CAP-treated and vehicle. The control group remained without any treatment. 0.5 mg/kg CAP prepared in a solvent and injected subcutaneously to CAP-treated group and vehicle group was injected with only solvent for 20 days. All rats were weighed daily. At the end of the experiment, tissue samples were collected and Crossman’s triple staining was used for histomorphometric examinations and labelled streptavidin-biotin immunohistochemical technique was used for SP expression. The body weight gain was lower in the CAP treated group but no significant differences between the groups. Low dose CAP did not alter the depth of crypt in the duodenum but the height of the villus was higher than the other group (p<0.05). In the CAP treated group, SP immunoreaction was stronger than the other groups in the surface epithelium, Lieberkühn crypts, in the Brunner’s glands, smooth muscles layer, and the neurons of the myenteric plexus of the duodenum. Finally, low dose CAP, markedly stimulated expression of SP and also induced positively histomorphometric changes in the duodenum. As a result of low dose CAP treatment may play important role in epithelial cell functions and in the regulation of digestive system physiology

Keywords

• Capsaicin
• duodenum
• immunohistochemistry
• rat
• substance P

Düşük doz kapsaisinin rat duodenumunda substans P ekspresyonu üzerine etkileri

Abstract

Sunulan çalışmanın amacı, düşük doz kapsaisinin (CAP) duodenumun histomorfometrik yapısı ve substans P (SP) ekspresyonu üzerine etkisini incelemektir. Çalışmada kullanılan ratlar (21 günlük), kontrol grubu, CAP enjekte edilen grup ve çözücü solüsyon enjekte edilen olmak üzere üç gruba ayrıldı. Kontrol grubuna enjeksiyon yapılmadı. 20 gün boyunca CAP grubuna çözücü solüsyon içerisinde hazırlanan 0.5 mg/kg kapsaisin, taşıyıcı grubuna ise sadece çözücü solüsyon subkutan enjekte edildi. Ratlar her gün tartıldı. Deney sonrası dokular alınarak, doku kesitlerine histomorfometrik incemeler için Crossman’ın üçlü boyaması, SP ekspresyonunu belirlemek için ise işaretlenmiş streptavidin-biotin immunohistokimyasal tekniği uygulandı. CAP uygulanan grupta canlı ağırlık kazancı kontrol gruplarına göre düşüktü, fakat gruplar arasında istatiksel farklılık gözlenmedi. Düşük doz CAP duodenumun kript derinliğini etkilemezken, villus uzunluğu CAP uygulanan grupta diğer gruplara göre daha yüksek belirlendi (p<0.05). SP immunreaksiyonu duodenumun yüzey epitelinde, Lieberkühn kriptlerinde, ve Brunner bezlerinde, diğer gruplara göre CAP grubunda daha şiddetli gözlendi. Sonuç olarak düşük doz CAP, SP ekspresyonunu arttırdı ve duodenumda olumlu histomorfometrik değişiklere sebep oldu. Düşük doz CAP uygulaması sonucu artan SP ekspresyonu sindirim fizyolojisinin düzenlenmesinde önemli rol oynayabilir

Keywords

• Duodenum
• immunohistokimya
• kapsaisin
• rat
• substans P

References

1. 1. Abdel-Salam OME, Debreceni A, Mózsik GY, et al. (1999): Capsaicin-sensitive afferent sensory nerves in modulating gastric mucosal defense against noxious agents. J Physiol, 93, 443-454.
2. 2. Abdel-Salam OME, Szolcsányi J, BarthóL, et al. (1994): Sensory nerve-mediated mechanisms, gastric mucosal damage and its protection: A critical overview. Gastroprotection, 2, 4-12.
3. 3. Ansel JC, Kaynard AH, Armstrong CA, et al. (1996): Skin-nervous system interactions. J Invest Dermatol, 106, 198-204.
4. 4. Brodin E, Rosen A, Schott E, et al. (1994): Effects of sequential removal of rats from a group cage, and of individual housing of rats, on substance P, cholecystokinin and somatostatin levels in the periaqueductal grey and limbic regions. Neuropeptides, 26, 253-60.
5. 5. Burbach GJ, Kim KH, Zivony AS, et al. (2001): The neurosensory tachykinins substance P and neurokinin A directly induce keratinocyte nerve growth factor. J Invest Dermatol, 117, 1075-1082.
6. 6. Caterina MJ, Schumacher MA, Tominaga M, et al. (1997): The capsaicin receptor: A heat activated ion channel in the pain pathway. Nature, 389, 816-824.
7. 7. Chang MM, Leeman SE (1970): Isolation of asialogogic peptide from bovine hypothalamic tissue and its characterization as substance P. J Biol Chem, 245, 4784- 4790.
8. 8. Chard PS, Bleakman D, Savidge JR, et al. (1995): Capsaicin-induced neurotoxicity in cultured dorsal root ganglion neurons: Involvement of calcium-activated proteases. Neuroscience, 65, 1099-1108.

9. 9. Erin N, Clawson GA (2004): Parameters affecting substance P measurement in heart, lung, and skin. Biotechnique, 232, 234-236.
10. 10. Erin N, Zik B, Sarigül M, et al. (2009): The effects of chronic low-dose capsaicin treatment on substance P levels. Regul Pept, 153, 83-87.
11. 11. Faussone-Pellegrini MS, Taddei A, Bizzoco E, et al. (2005): Distribution of the vanilloid (capsaicin) receptor type 1 in the human stomach. Histochem Cell Biol, 124, 61-68.
12. 12. Geppetti P, Trevisani M (2004): Activation and sensitization of the vanilloid receptor: Role in gastrointestinal inflammation and function. Br J Pharmacol, 141, 1313- 1320.
13. 13. Grady EF, Baluk P, Bohm S, et al. (1996): Characterization of antisera specific to NK1, NK2, and NK3 neurokinin receptors and their utilization to localize receptors in the rat gastrointestinal tract. J Neurosci, 16, 6975-6986.
14. 14. Green T, Dockray GJ (1987): Calcitonin gene-related peptide and substance P in afferents to the upper gastrointestinal tract in the rat. Neurosci Lett, 76, 151-156.
15. 15. Hoffmann P, Mazurkiewicz J, Holtmann G, et al. (2002): Capsaicin-sensitive nerve fibres induce epithelial cell proliferation, inflammatory cell immigration and transforming growth factor-alpha expression in the rat colonic mucosa in vivo. Scand J Gastroenterol, 37, 414-422.
16. 16. Holzer P, Sametz W (1986): Gastric mucosal protection against ulcerogenic factors in the rat mediated by capsaicin-sensitive afferent neurons. Gastroenterology, 91, 975-981.
17. 17. Holzer-Petsche U, Seitz H, Lembeck F (1989): Effect of capsaicin on gastric corpus smooth muscle of the rat in vitro. Eur J Pharmacol, 162, 29-36.
18. 18. Holzer P (1991): Capsaicin: Cellular targets, mechanisms of action, and selectivity for thin sensory neurons. Pharmacol Rev, 43, 143-201.
19. 19. Holzer P, Livingston EH, Saria A, et al. (1991): Sensory neurons mediate protective vasodilation in rat gastric mucosa. Am J Physiol, 260, 363-370.
20. 20. Holzer P (1998): Neural emergency system in the stomach. Gastroenterology, 114, 823-839.
21. 21. Höckerfelt U, Franzon L, Forsgren S (2001): Substance P (NK1) receptor in relation to substance P innervation in rat duodenum after irradiation. Regul Pept, 98, 115-126.
22. 22. Huang ZL, Mochizuki T, Watanabe H, et al. (1999): Activation of sensory nerves participates in stress-induced histamine release from mast cells in rat. Neurosci Lett, 270, 181-184.
23. 23. Jackson P, Blythe D (1993): Immunolabelling techniques for light microscopy. 15-43 In: Beesley JE (Ed), Immunocytochemistry. Oxford University Press, New York.
24. 24. Kang JY, Alexander B, Math MV, et al. (1993): The effect of chilli and its pungent ingredient capsaicin on gastrointestinal transit in the rat. J Gastroenterol Hepatol, 8, 513-516.
25. 25. Kang JY, Teng CH, Wee A, et al. (1995): Effect of capsaicin and chilli on ethanol induced gastric mucosal injury in the rat. Gut, 36, 664-669.
26. 26. Kawada T, Hagihara KI, Iwai K (1986): Effects of capsaicin on lipid metabolism in rats fed a high fat diet. Journal Nutr, 116, 1272-1280.
27. 27. Keast JR, Furness JB, Costa M (1985): Distribution of certain peptide containing nerve fibers and endocrine cells in the gastrointestinal mucosa of five mammalian species. J Comp Neurol, 236, 403-422.
28. 28. Kempaiah RK, Srinivasan K (2004): Influence of dietary curcumin, capsaicin and garlic on the antioxidant status of red blood cells and the liver in high fat fed rats. Ann Nutr Metab, 48, 314-320.
29. 29. Kim C, Robert AM, Joshua IG (1995): Differences in the capsaicin-induced dilation of arterioles and venules in rat striated muscle. J Pharmacol Exp Ther, 273, 605-610.
30. 30. Lippe IT, Pabst MA, Holzer P (1989): Intragastric capsaicin enhances rat gastric acid elimination and mucosal blood flow by afferent nerve stimulation. Br J Pharmacol, 96, 91-100.
31. 31. Lobb D (2009): Expression and actions of transforming growth factors during human follicular development. Fertil Steril, 92, 1080-1084.
32. 32. Maggi CA, Patacchini R, Giachetti A, et al. (1990): Tachykinin receptors in the circular muscle of the guineapig ileum. Br J Pharmacol, 101, 996-1000.
33. 33. Mathison R, Davison JS (1989): Regulation of epithelial ion transport in the jejunal mucosa of the guinea pig by neurokinins. Life Sci, 45, 1057-1064.
34. 34. Moore BA, Kim D, Vanner S (2000): Neural pathways regulating Brunner's gland secretion in guinea pig duodenum in vitro. Am J Physiol Gastrointest Liver Physiol, 279, 910-917.
35. 35. Naponitaya W (1973): Long-term effects of capsaicin on fat absorption and the growth of the rat. Growth, 37, 269- 279.
36. 36. Ozer A, Zik B, Erdost H, et al. (2006): Histological investigations on the effects of feeding with a diet containing red hot pepper on the reproductive system organs of the cock. Turk J Vet Anim Sci, 30, 7-15.
37. 37. Park JS, Choi MA, Kim BS, et al. (2000): Capsaicin protects against ethanol-induced oxidative injury in the gastric mucosa of rats. Life Sci, 67, 3087-3093.
38. 38. Parsons AM, Seybold VS, Chandan R, et al. (1992): Neurokinin receptors and mucosal ion transport in porcine jejunum. J Pharmacol Exp Ther, 261, 1213-1221.
39. 39. Pothoulakis C, Castagliuolo I, Leeman SE, et al. (1998): Substance P receptor expression in intestinal epithelium in Clostridium difficile toxin A enteritis in rats. Am J Physiol, 275, 68-75.
40. 40. Reid TW, Murphy CJ, Iwahashi CK, et al. (1993): Stimulation of epithelial cell growth by the neuropeptide substance P. J Cell Biochem, 52, 476-485.
41. 41. Renzi D, Pellegrini B, Tonelli F, et al. (2000): Substance P (neurokinin-1) and neurokinin A (neurokinin-2) receptor gene and protein expression in the healthy and inflamed human intestine. Am J Pathol, 157, 1511-1522.
42. 42. Shanahan F (1998): Enteric neuropathophysiology and inflammatory bowel disease. Neurogastroenterol Motil, 10, 185-187.
43. 43. Sharkey KA, Williams RG, Dockray GJ (1984): Sensory substance P innervation of the stomach and pancreas. Demonstration of capsaicin-sensitive sensory neurons in the rat by combined immunohistochemistry and retrograde tracing. Gastroenterology, 87, 914-21.
44. 44. Sokolski KN, Lechago J (1984): Human colonic substance P-producing cells are a separate population from the serotonin-producing enterochromaffin cells. J Histochem Cytochem, 32, 1066-1074.
45. 45. Srinivasan MR, Satyanarayana MN (1989): Effect of capsaicin on skeletal muscle lipoprotein lipase in rats fed high fat diet. Indian J Exp Biol, 27, 910-2.
46. 46. Sundler F, Hakanson R, Larsson LI, et al. (1977): Substance P in the gut: Immunohistochemical and immunocytochemical study of distribution and development. 59–65, In: von Euler US, Pernow B (Eds), Substance P. Raven Press, New York.
47. 47. Szolcsanyi J, Bartho L (2001): Capsaicin-sensitive afferents and their role in gastroprotection: an update. J Physiol, 95, 181-188.
48. 48. Takeuchi K, Ueshima K, Matsumoto J, et al. (1992): Role of capsaicin- sensitive sensory nerves in acid-induced bicarbonate secretion in rat 
stomach. Dig Dis Sci, 37, 737-743.
49. 49. Tutuncu S, Ozfiliz O (2011): Distribution of the vanilloid (capsaicin) receptor type 1 in the capsaicin treated rat ovaries on different sexual development periods. Revue Méd Vét, 162, 460-467.
50. 50. Udupihille M (1993): The effect of capsaicin on the small intestinal absorption of glucose and alanine in the rat. Indian J Physiol Pharmacol, 37, 59-62.
51. 51. Wang JX, Peng KM (2008): Developmental morphology of the small intestine of African ostrich chicks. Poult Sci, 87, 2629-2635.
52. 52. Zik B, Ozguden C, Tutuncu S et al. (2007): Expression of vanilloid receptor-1 in the duodenum of the capsaicin treated rat. Bull Vet Inst Pulawy, 51, 149-153.