Histochemical investigations of the effects of Cyclamen graecum extracts on the rat lower gastrointestinal system

Abstract

Aim: It is known that saponins, triterpenoid or steroidal glycosides, have effects on gastrointestinal system (GIS) and mucus. Cyclamen graecum, a tuberous plant belonging to the Primulaceae family, contains cyclamin as the main saponin. In the present study, it was aimed to investigate the effects of C. graecum tubers ethanolic extracts at the concentrations of 0.1 and 0.3 g/L on the rat lower GIS histomorphology and mucus intensity. Material-Method: In this study, rats were allocated to 3 groups, each comprising 5 rats [Group I: control group, physiologic saline was administered; Group II: Experimental group, 0.1 g/L(aq) C. graecum extract was administered; Group III: Experimental group, 0.3 g/L(aq) C. graecum extract was administered]. C. graecum tubers ethanolic extracts were given orally to the rats during 10 weeks. At the end of the experimental period, Hematoxylin and Eosin (H&E), Periodic acid-Schiff (PAS), Alcian Blue (AB) pH 1 and AB pH 2.5 histochemical staining techniques were employed on the tissue samples of small and large intestines in order to demonstrate the histomorphological structures, the neutral, sulphated and acidic mucosubstances, respectively. Results: In both experimental groups, small and large intestine histomorphology, and AB and PAS positive reaction intensities of neutral, acidic, and sulphated mucosubstances were similar to control group. Conclusion: It can be suggested that the C. graecum ethanolic extract has no effects on the rat lower GIS histomorphology and mucus intensity at the doses administered.

Keywords

• Cyclamen graecum, lower gastrointestinal system, mucus, histochemistry

Cyclamen graecum ekstraktlarının sıçan alt gastrointestinal sistem üzerindeki etkilerinin histokimyasal olarak araştırılması

Abstract

Amaç: Steroidal veya triterpenoid glikozitler olan saponinlerin, gastrointestinal sistem (GİS) ve mukus üzerinde etkilerinin olduğu bilinmektedir. Primulaceae familyasına ait yumrulu bir bitki olan Cyclamen graecum, başlıca saponin olarak siklamini içerir. Bu çalışmada, C. graecum yumrularının 0,1 ve 0,3 g/l konsantrasyonlarındaki etanolik ekstraktlarının, sıçan alt GİS histomorfolojisi ve mukus yoğunluğu üzerindeki etkilerinin araştırılması amaçlanmıştır. Materyal-Metot: Çalışmada, her biri 5 sıçan içeren 3 grup [Grup I: Fizyolojik su verilen kontrol grubu, Grup II: 0,1 g/l (suda) C. graecum ekstraktı verilen deney grubu, Grup III: 0,3 g/l (suda) C. graecum ekstraktı verilen deney grubu] oluşturulmuştur. C. graecum etanolik ekstraktları 10 hafta süresince hayvanlara oral yolla verilmiştir. Deney sonunda, ince ve kalın bağırsak doku örneklerine, histomorfolojik yapıyı göstermek için Hematoksilen-Eosin (H&E), nötral mukus maddesi için Periyodik Asit Schiff (PAS), sülfatlı mukus maddesi için Alsiyan Mavisi (AB) pH 1 ve asidik mukus maddesi için AB pH 2,5 histokimyasal boyama yöntemleri uygulanmıştır. Bulgular: Her iki deney grubunda, ince ve kalın bağırsak histomorfolojisi ile nötral, asidik ve sülfatlı mukus maddesi AB ve PAS pozitif reaksiyon yoğunlukları kontrol grubu ile benzerlik göstermiştir. Sonuç: C. graecum etanolik ekstraktının uygulanan konsantrasyonlarının, sıçan alt GİS histomorfolojisi ve mukus yoğunluğu üzerinde bir değişikliğe neden olmadığı söylenebilir.

Keywords

• Cyclamen graecum, alt gastrointestinal sistem, mukus, histokimya

References

1. Allen A, Hutton DA, Pearson JP, Sellers LA. Mucus glycoprotein structure, gel formation and gastrointestinal mucus function. Ciba Found Symp. 1984; 109: 137-156.
2. Juge N. Microbial adhesins to gastrointestinal mucus. Trends Microbiol 2012; 20(1):30-39.
3. Mackie R, Round AN, Rigby NM, Macierzanka A. The role of the mucus barrier in digestion. Food Dig 2012; 3(1-3): 8-15.
4. Hansson GC. Role of mucus layers in gut infection and inflammation. Curr Opin Microbiol 2012; 15(1): 57-62.
5. Morrissey SM, Ward PM, Jayaraj AP, Tovey FI, Clark CG. Histochemical changes in mucus in duodenal ulceration. Gut 1983; 24: 909-913.
6. Nikumbh RD, Nikumbh DB, Umarji BN. Mucin histochemical study of the colon in normal and malignant lesions. Int J Health Sci Res. 2012; 2(7): 20-32.
7. Gomes J, Magalhães A, Carvalho AS, Hernandez GE, Papp SL, Head SR, Michel V, David L, Gärtner F, Touati E, Reis CA. Glycophenotypic alterations induced by Pteridium aquilinum in mice gastric mucosa: synergistic effect with Helicobacter pylori infection. 2012; PLoS ONE 7(6): e38353. doi:10.1371/journal.pone. 0038353.
8. Singh P, Guha D. Aegle marmelos enhances gastric mucosal protection: relevance for NSAIDS-induced gastric mucosal injury. Al Ame en J Med Sci 2012; 5(3): 243-255.

9. Keskin N, Ili P, Sahin B. Histochemical demonstration of mucosubstances in the mouse gastrointestinal tract treated with Origanum hypericifolium O. Schwartz and P.H. Davis extract. Afr J Biotechnol 2012; 11(10): 2436-2444.
10. Wina E, Muetzel S, Becker K. The impact of saponins or saponin-containing plant materials on ruminant production: a review. J Agric Food Chem 2005; 53(21): 8093-8105.
11. Osbourn A. Saponins and plant defence-a soap story. Trends Plant Sci 1996; 1(1): 1-9.
12. Akbel E, Karabağ F. Saponinler ve reproduktif etkileri. Türk Bilimsel Derlemeler Dergisi 2012; 5(1): 25-29.
13. http://nasodrensinusitis.blogspot.com/2012/03/cyclamen- europaeum-extract.html.
14. Pfaar O, Mullol J, Anders C, Hörmann K, Klimek L. Cyclamen europaeum nasal spray, a novel phytotherapeutic product for the management of acute rhinosinusitis: a randomized double-blind, placebo-controlled trial. Rhinology 2012; 50(1): 37-44.
15. Harvala C, Hylands PJ. Saponins from Cyclamen hederifolium and C. graecum. Planta Med 1978; 33(2): 180- 184.
16. Feresin GE, Tapia AA, Bustos DA. Antibacterial activity of some medicinal plants from San Juan, Argentina. Fitoterapia 2000; 71(4): 429-432.
17. Sainte-Marie GA. Paraffin embedding technique for studies employing immunofluorescence. J Histochem Cytochem 1962; 250-256.
18. Harris HF. A new method of ripening haematoxylin. Micr Bull (Philadelphia). 1898; 47.
19. McManus JFA. Histological and histochemical uses of periodic acid. Stain Tech. 1948; 23: 99-108.
20. Lev R, Spicer SS. Specific staining of sulphate groups with alcian blue at low pH. J Histochem Cytochem 1964; 12: 309.
21. Mowry RW. Alcian blue techniques for the histochemical study of acidic carbohydrates. J Histochem Cytochem 1956; 4: 407-408.
22. Gee JM, Johnson IT. Interactions between hemolytic saponins, bile salts and small intestinal mucosa in the rat. J Nutr 1988; 118(11): 1391-1397.
23. Bureau DP, Harris AM, Cho CY. The effects of purified alcohol extracts from soy products on feed intake and growth of chinook salmon (Oncorhynchus tshawytscha) and rainbow trout (Oncorhynchus mykiss). Aquaculture 1998; 161(1-4): 27-43.
24. Iwashita Y, Yamamoto T, Furuita H, Sugita T, Suzuki N. Influence of certain soybean antinutritional factors supplemented to a casein-based semipurified diet on intestinal and liver morphology in fingerling rainbow trout Oncorhynchus mykiss. Fish Sci 2008; 74(5): 1075-1082.
25. Akinpelu BA, Oyedapo OO, Iwalewa EO, Shode F. Biochemical and histopathological profile of toxicity induced by saponin fraction of Erythrophleum suaveolens (Guill. & Perri.) bark extract. Phytopharmacology 2012; 3(1): 38-53.
26. Osuigwe DI, Obiekezie AI, Onuoha GC. Effects of jackbean seed meal on the intestinal mucosa of juvenile Heterobranchus longifilis. Afr J Biotechnol 2006; 5(13): 1294-1298.
27. Badombena-Wanta DB, Metowogo K, Tettegah M, Lawson-Evi P, Eklu-Gadegbeku K, Aklikokou AK, Gbeassor M. Comparative anti-ulcers effect of hydroalcoholic extract of leaves and root of Calotropis procera Ait (Asclepiadaceae). Res J Pharm Biol Chem Sci 2013; 4(3): 1205-1212.
28. Owoyele BV, Gbago AF, Ashaolu OS. Gastroprotective effects of aqueous extract of unripe Carica papaya fruit in rats. Pac J Med Sci 2013; 11(2): 3-11.
29. Marhuenda E, Martin MJ, Alarcon Lastra CDL. Antiulcerogenic activity of aescine in different experimental models. Phytother Res 1993; 7: 13-16.
30. Marhuenda E, Alarcón de la Lastra C, Martín MJ. Antisecretory and gastroprotective effects of aescine in rats. Gen Pharmacol-Vasc S 1994; 25(6): 1213-1219.
31. Lee S, Kim MG, Ko SK, Kim HK, Leem KH, Kim YJ. Protective effect of ginsenoside Re on acute gastric mucosal lesion induced by compound 48/80. J Ginseng Res 2014; 38(2): 89-96.