An Immunohistochemical Investigation of The Effect of Sambucus Nigra on Chymase-, Tryptase- and Ghrelin- Positive Cells in Rat Lung

Öz

Sambucus nigra (S.nigra) is used in the treatment of many diseases and disorders thanks to its antioxidant, anticarcinogenic, immunostimulating, antiallergic, antiviral, and antibacterial properties. Ghrelin has anti-inflammatory effect on oxidative damage in various organs and cell types. The aim of this study was to immunohistochemically examine the chymase, tryptase, and ghrelin in rat lung after S. nigra administration. A total of 16 male rats were used in the study. The rats were assigned to two groups, control and S. nigra. 1st control group (n=8): No application was made. 2nd S. nigra group (n=8): S. nigra extract was administered at 15 mg/kg by oral gavage for 14 days. Tryptase, chymase, and ghrelin-positive cells were found in the lung tissue in a spindle-shaped, round, or oval shape. When the groups were evaluated within themselves, a significant increase in the number of tryptase, chymase, and ghrelin positive cells was observed in the S. nigra treated group. This study showed that S. nigra, which has an immunomodulatory and antioxidant effect, increases the expression of chymase-, tryptase- and ghrelin-positive cells in the lungs. Additionally, based on our findings, it can be said that mast cells can produce, store and release ghrelin.

Anahtar Kelimeler

• Chymase
• ghrelin
• lung
• Sambucus nigra
• tryptase

Sıçan Akciğerinde Sambucus Nigra’nın Kimaz, Triptaz ve Ghrelin Pozitif Hücrelerin Üzerindeki Etkisinin İmmünohistokimyasal Olarak İncelenmesi

Öz

Sambucus nigra (S. nigra) antioksidan, antikanserojenik, immün sistemi uyarıcı, antialerjik, antiviral ve antibakteriyel özellikleri sayesinde birçok hastalık ve rahatsızlığın tedavisinde kullanılmaktadır. Ghrelin, çeşitli organlarda ve hücre tiplerinde oksidatif hasar üzerinde anti-inflamatuar etkilere sahiptir. Bu çalışmanın amacı, S. nigra uygulamasından sonra sıçan akciğerinde kimaz, triptaz ve ghrelin immünopozitif hücrelerinin immünohistokimyasal olarak incelenmesidir. Çalışmada toplam 16 erkek sıçan kullanıldı. Sıçanlar kontrol ve S. nigra olmak üzere iki gruba ayrıldı. 1. kontrol grubu (n=8): herhangi bir uygulama yapılmadı. 2. S. nigra grubu (n=8): S. nigra ekstresi 14 gün süreyle oral gavaj yoluyla 15 mg/kg dozunda uygulandı. Akciğer dokusunda yuvarlak, oval veya mekik şeklinde triptaz, kimaz ve ghrelin pozitif hücreler bulundu. İki grup kendi içinde değerlendirildiğinde S. nigra uygulanan grupta triptaz, kimaz ve ghrelin pozitif hücre sayısında önemli artış gözlendi. Bu çalışma, immünomodülatör ve antioksidan etkiye sahip olan S. nigra'nın akciğerde kimaz, triptaz ve ghrelin pozitif hücrelerin ekspresyonunu arttırdığını göstermiştir. Ayrıca mast hücrelerinin diğer bağışıklık hücreleri gibi ghrelin üretebildiği, depolayabildiği ve serbest bırakabildiği açıklanmaya çalışılmıştır.

Anahtar Kelimeler

• Akciğer
• ghrelin
• kimaz
• Sambugus nigra
• triptaz

Kaynakça

1. Akalu Y, Molla MD, Dessie G, Ayelign B. Physiological effect of ghrelin on body systems. Int J Endocrinol. 2020; 25:1385138. doi: 10.1155/2020/1385138.
2. Baatar D, Patel K, Taub DD. The effects of ghrelin on inflammation and the immune system. Mol Cell Endocrinol. 2011; 340(1): 44-58. doi: 10.1016/j.mce.2011.04.019.
3. Bidian C, Mitrea DR, Tatomir C, Perdeschrepler M, Lazăr C, Chetan I, Bolfa P, David L, Clichici S, Filip GA, Mureșan M, Micle O. Vıtıs Vınıfera L. And Sambucus Nıgra L. extracts attenuate oxıdatıve stress and ınflammatıon ın femoral ıschemıa. Farmacia. 2021; 69:1. doi.org/10.31925/farmacia.2021.1.8.
4. Chen J, Liu XJ, Shuo QL, Li SQ, Luo FM. Ghrelin attenuates lipopolysaccharide-induced acute lung injury through no pathway. Med Sci Monitor. 2008; 14(7): 141-146.
5. Chowen JA, Argente J. Ghrelin: A Link Between Energy Homeostasis and the Immune System. Endocrinology. 2017; 158(7): 2077-2081. doi: 10.1210/en.2017-00350.
6. Çetinel S, Çanıllıoğlu YE, Çikler E, Sener G, Ercan F. Leukotriene D4 receptor antagonist montelukast alleviates protamine sulphate-induced changes in rat urinary bladder. BJU Int. 2011; 107(8): 1320-1325. doi: 10.1111/j.1464-410X.2010.09532.x.
7. Da Silva EZM, Jamur MC, Oliver C. Mast cell function: a new vision of an old cell. J Histochem Cytochem. 2014; 62(10): 698-738. doi: 10.1369/0022155414545334.
8. Dell'Italia LJ, Husain A. Dissecting the role of chymase in angiotensin II formation and heart and blood vessel diseases. Curr Opin Cardiol. 2002; 17(4): 374-379. doi: 10.1097/00001573-200207000-00009.

9. Dwyer DF, Barrett NA, Austen KF. Expression profiling of constitutive mast cells reveals a unique identity within the immune system. Nat Immunol. 2016; 17(7):878-887. doi: 10.1038/ni.3445.
10. Duymuş HG, Göger F, Başer KHC. In vitro antioxidant properties and anthocyanin compositions of elderberry extracts. Food Chem. 2014; 155: 112-119. doi: 10.1016/j.foodchem.2014.01.028.
11. Ertuğrul T, Tutuncu Ş, Ozdemır B, Delıce N. Possible effect of thymoquinone on mast cell number and chymase, IL-4 and IFN-γ expression in rat spleen. Med Weter 2021; 77(10): 484-490. doi.org/10.21521/mw.6580.
12. Fazio A, Plastina P, Meijerink J, Witkamp RF, Gabriele B. Comparative analyses of seeds of wild fruits of Rubus and Sambucus species from southern Italy: fatty acid composition of the oil, total phenolic content, antioxidant and anti-inflammatory properties of the methanolic extracts. Food Chem. 2013; 140(4): 817-824. doi: 10.1016/j.foodchem.2012.11.010.
13. Gnanapavan S, Kola B, Bustin SA, Morris DG, McGee P, Fairclough P, Bhattacharya S, Carpenter R, Grossman AB, Korbonits M. The tissue distribution of the mRNA of ghrelin and subtypes of its receptor, GHS-R, in humans. J Clin Endocrinol Metab. 2002; 87(6): 2988. doi: 10.1210/jcem.87.6.8739.
14. Hamada H, Terai M, Kimura H, Hirano K, Oana S, Niimi H. Increased expression of mast cell chymase in the lungs of patients with congenital heart disease associated with early pulmonary vascular disease. Am J Respir Crit Care Med. 1999; 160(4): 1303-1308. doi: 10.1164/ajrccm.160.4.9810058.
15. Hawkins J, Baker C, Cherry L, Dunne E. Black elderberry (Sambucus nigra) supplementation effectively treats upper respiratory symptoms: a metaanalysis of randomized, controlled clinical trials. Complement Ther Med. 2019; 42:3 61-65. doi.org/10.1016/j.ctim.2018.12.004.
16. Hayıroğlu AE, Karaca T, Demirtaş S. Streptozotosin ile deneysel diyabet oluşturulan sıçanlarda östrus siklusunun değişik evrelerinde ovaryum ve uterus dokularında mast hücrelerinin dağılımının histokimyasal ve immünohistokimyasal olarak incelenmesi. Kafkas J Med Sci. 2016; 6(1):29–37. doi: 10.5505/kjms.2016.30074.
17. Hirata K, Sugama Y, Ikura Y, Ohsawa M, Inoue Y, Yamamoto S, Kitaichi M, Ueda M. Enhanced mast cell chymase expression in human idiopathic interstitial pneumonia. Int J Mol Med. 2007; 19(4): 565-570.
18. Ivanova K, Stefanov I, Ivanova I, Ananiev J, Gulubova M. Ghrelın expressıon in mast cells of infant lung with respıratory distress syndrome. Acta Med. Bulg. 2021; 48: 40-45. doi.org/10.2478/amb-2021-0006.
19. Ivanova IG, Stefanov IS. Tryptase- and ghrelin positive mast cells in the interalveolar septa of rat’s lung. Bulg J Vet Med. 2021; 24(4): 469-477.
20. Kojima M, Kangawa K. Ghrelin: structure and function. Physiol Rev. 2005; 85(2): 495-522. doi: 10.1152/physrev.00012.2004.
21. Kosanovic D, Luitel H, Dahal BK, Cornitescu T, Janssen W, Danser AH, Garrelds IM, De Mey JGR, Fazzi G, Schiffers P, Iglarz M, Fischli W, Ghofrani HA, Weissmann N, Grimminger F, Seeger W, Reiss I, Schermuly RT. Chymase: a multifunctional player in pulmonary hypertension associated with lung fibrosis. Eur Respir J. 2015; 46(4): 1084-1094. doi: 10.1183/09031936.00018215.
22. Li M, Yang K, Wang X, Xu X, Zhu L, Wang H. Mast cells infiltration and decreased E-cadherin expression in ketamine-induced cystitis. Toxicol Rep. 2014; 8(2): 205-209. doi: 10.1016/j.toxrep.2014.12.003.
23. Metcalfe DD, Baram D, Mekori YA. Mast cells. Physiol Rev. 1997; 77(4):1033-1079. doi: 10.1152/physrev.1997.77.4.1033.
24. Miyazaki M, Takai S, Jin D, Muramatsu M. Pathological roles of angiotensin II produced by mast cell chymase and the effects of chymase inhibition in animal models. Pharmacol Ther. 2006; 112(3):668-676. doi: 10.1016/j.pharmthera.2006.05.008.
25. Mitani Y, Ueda M, Maruyama K, Shimpo H, Kojima A, Matsumura M, Aoki K, Sakurai M. Mast cell chymase in pulmonary hypertension Thorax. 1999; 54(1): 88-90. doi: 10.1136/thx.54.1.88.
26. Mukai K, Tsai M, Saito H, Galli SJ. Mast cells as sources of cytokines, chemokines, and growth factors. Immunol Rev. 2018; 282(1): 121-150. doi: 10.1111/imr.12634.
27. Nakazato M, Murakami N, Date Y, Kojima M, Matsuo H, Kangawa K, Matsukura S. A rolefor ghrelin in the central regulation of feeding. Nature. 2001; 409(6817):194-8. doi: 10.1038/35051587.
28. Narin N, Çetin E. Effect of ghrelin administration on some hematological parameters in rats. J Health Sci. 2010; 19(3): 202-208.
29. Oniszczuk A, Olech M, Oniszczuk T, Wojtunik-Kulesza K, Wójtowicz A. Extraction methods, LC-ESI-MS/MS analysis of phenolic compounds and antiradical properties of functional food enriched with elderberry flowers or fruits. Arabian Journal of Chemistry. 2016; 12(8): 4719-4730. doi.org/10.1016/j.arabjc.2016.09.003.
30. Payne V, Kam PC. Mast cell tryptase: a review of its physiology and clinical significance. Anaesthesia 2004; 59(7): 695-703. doi: 10.1111/j.1365-2044.2004.03757.x.
31. Pejler G, Ronnberg E, Waern I, Wernersson S. Mast cell proteases: multifaceted regulators of inflammatory disease. Blood. 2010; 115(24): 4981-4990. doi: 10.1182/blood-2010-01-257287.
32. Raghay K, Akki R, Bensaid D, Errami M. Ghrelin as an anti-inflammatory and protective agent in ischemia/ reperfusion injury. Peptides. 2020; 124:170226. doi: 10.1016/j.peptides.2019.170226.
33. Sakaguchi M, Takai S, Jin D, Okamoto Y, Muramatsu M, Kim S, Miyazaki M. A specific chymase inhibitor, NK3201, suppresses bleomycin-induced pulmonary fibrosis in hamsters. Eur J Pharmacol. 2004; 493(1-3): 173-176. doi: 10.1016/j.ejphar.2004.04.024.
34. Samrao D, Wang D, Ough F, Lin YG, Liu S, Menesses T, Yessaian A, Turner N, Pejovic T, Mhawech-Fauceglia P. Histologic parameters predictive of disease outcome in women with advanced stage ovarian carcinoma treated with neoadjuvant chemotherapy. Transl Oncol. 2012; 5(6): 469-474. doi: 10.1593/tlo.12265.
35. Schwartz LB, Lewis RA, Seldin D, Austen KF. Acid hydrolases and tryptase from secretory granules of dispersed human lung mast cells. J Immunol. 1981; 126(4): 1290-1294.
36. Schwenke DO, Tokudome T, Shirai M, Hosoda H, Horio T, Kishimoto, Kangawa K. Exogenous ghrelin attenuates the progression of chronic hypoxia-induced pulmonary hypertension in conscious rats. Endocrinology. 2008; 149(1): 237-44. doi: 10.1210/en.2007-0833.
37. Sonneck K, Florian S, Böhm A, Krauth MT, Kondo R, Hauswirth AW, Gleixner KV, Aichberger KJ, Derdak S, Pickl WF, Sperr WR, Schwartz LB, Valent P. Evaluation of biologic activity of tryptase secreted from blast cells in acute myeloid leukemia. Leuk Lymphoma. 2006; 47(5): 897-906. doi: 10.1080/10428190500513652.
38. Stefanov IS, Ananiev JR, Ivanovab KV, Tolekovac AN, Vodenicharovd AP, Gulubova MV. Distribution of ghrelin-positive mast cells in rat stomach. Biotechnol Biotechnol Equip. 2017; 31(4): 774–781. doi.org/10.1080/13102818.2017.1326013.
39. Stefanov I. Mast cell distribution in porcine common bile duct with special reference to ghrelin. Bulg J Vet Med. 2021; (online first). doi: 10.15547/bjvm.2020-0127.
40. Stefanov I, Vodenicharov A, Atanassova P, Hrischev P, Vulkova I, Stoyanov D, Tsandev N, Hristov HA. Mast cell density in domestic swine common hepatic duct. Bulg. J. Vet. Med. 2021; (online first). doi: 10.15547/bjvm.2020-0145.
41. Tomimori Y, Muto T, Saito K, Tanaka T, Maruoka H, Sumida M, Fukami H, Fukuda Y. Involvement of mast cell chymase in bleomycin-induced pulmonary fibrosis in mice. Eur J Pharmacol. 2003; 478(2-3): 179-185. doi: 10.1016/j.ejphar.2003.08.050.
42. True LD. Principles of immunohistochemistry, Gower Medical Publishing, New York. 1990; p. 67-75.
43. Tutuncu S, Ertuğrul T. Immunohistochemical expression of tryptase-chymase and mast cell heterogeneity in capsaicin-treated rat ovaries. IJVAR. 2019; 2(2): 25-31.
44. Tütüncü Ş, Torun AÇ, Ertuğrul T. The role of resveratrol on mast cell and chymase and tryptase expression in blunt-chesttrauma-induced acute lung injury in rats. Turk J Vet Anim Sci. 2020; 44: 1260-1268. doi:10.3906/vet-2005-23.
45. Xia Q, Pang W, Pan H, Zheng Y, Kang JS, Zhu SG. Effects of ghrelin on theproliferation and secretion of splenic T lymphocytes in mice. Regul Pept. 2004; 122(3):173-178. doi: 10.1016/j.regpep.2004.06.016.
46. Uranga JA, Martínez V, Abalo R. Mast cell regulation and ırritable bowel syndrome: effects of food components with potential nutraceutical use. Molecules. 2020; 25(18): 4314. doi: 10.3390/molecules25184314.
47. Van der Velden J, Barker D, Barcham G, Koumoundouros E, Snibson K. Increased mast cell density and airway responses to allergic and non-allergic stimuli in a sheep model of chronic asthma. PLoS One. 2012; 7(5): e37161. doi: 10.1371/journal.pone.0037161.
48. Volante M, Fulcheri E, Allìa E, Cerrato M, Pucci A, Papotti M. Ghrelin expression in fetal, infant, and adult human lung. J Histochem Cytochem. 2002; 50(8): 1013-1021. doi: 10.1177/002215540205000803.
49. Wieland LS, Piechotta V, Feinberg T, Ludeman E, Hutton B, Kanji S, Seely D, Garritty C. Elderberry for prevention and treatment of viral respiratory illnesses: a systematic review. BMC Complement Med Ther. 2021; 21(1): 112. doi: 10.1186/s12906-021-03283-5.
50. Zhao W, Gan X, Su G, Wanling G, Li S, Hei Z, Yang C, Wang H. The interaction between oxidative stress and mast cell activation plays a role in acute lung injuries induced by intestinal ischemia-reperfusion. J Surg Res. 2014; 187 (2): 542-552. doi.org/10.1016/j.jss.2013.10.033.