İnsan Umblikal Ven Endotel Hücre Kültüründe TNF- lfa İle Oluşturulmuş İn Vitro Greft Versus Host Hastalığı Modelinde Adrenomedullinin Sitoprotektif Etkisi

Year 2014, Volume: 11 Issue: 3, 210 - 216, 15.12.2014

Fatih Kurnaz Yasemin Torun Çiğdem Pala Esma Kaya Serdar Şıvgın Leylagül Kaynar , Bülent Eser Fatih Şahin Ahmet Öztürk Mustafa Çetin

Abstract

Giriş: Allojeneik kök hücre nakli (KHN) özellikle hematolojik malignitelerin tedavisinde önemli bir yer
tutmaktadır. Greft versus host hastalığı (GVHH) allojeneik KHN'nin en sık görülen komplikasyonlarından
biridir. Nakil sonrası GVHH'da temel patolojik mekanizmalardan birinin inflamatuar yolak üzerinden
endotel hücre hasarı olduğu düşünülmektedir. İnflamatuar yolak üzerinden koruyucu etkisi olan
adrenomedullin (AMD)'nin TNF-α ile oluşturulmuş in-vitro GVHH modelindeki sitoprotektif etkisi
araştırıldı. 

Methods: İnsan umblikal veninden elde edilen endotel hücreleri TNF-α (grup1) ve TNF-α+AMD (grup2) ile
iki saat inkübe edildi ve endotel hücre kaynaklı mikropartiküller (CD62, CD146, CD31, CD51/61) ve
apoptozis belirteci olan Anneksin-Völçüldü.
Bulgular: Her iki grupta endotelyal mikropartikül seviyeleri arasında fark yoktu. AMD sonrası bazı
ölçümlerde artış tespit edilirken bazı ölçümlerde düşüş tespit edildi (p>0,05).
Sonuç: In vitro akut GVHH modelinde AMD'in koruyucu etkisi olup olmadığıni tespit etmek için çok
ölçümlü ve büyük çalışmalara ihtiyaç vardır. 

Keywords

Adrenomedullin, greft versus host hastalığı, mikropartikül

Effects Of Adrenomedullin On Endothelial Microparticle Release In TNF-Αlpha Stimulated In-Vitro Graft Versus Host Disease Model

Abstract

Background: Allogeneic stem cell transplantation (SCT) is an important treatment modality used in
hematological malignant disorders. Graft-versus-host disease (GVHD) is the main complication after
allogeneic SCT. It is suggested that endothelial cell damage has an important role in development of GVHD.
In this study we investigated the effect of adrenomedullin (AMD) in TNF-α stimulated in-vitro GVHD model
since AMD has a protective effect on cells via the regulation inflammatory pathway.
Materials and Methods: Umbilical cord derived endothelial cells were incubated with TNF-α for 2 hours in
group1 and with TNF-α+AMD in group2 in order to make in-vitro GVHD model and see the effect of AMD.
Microparticles (CD62, CD146, CD31, CD51/61) originated from endothelial cells that are the markers of
injury or apoptosis were measured.
Results: We found no significant difference between the microparticle levels in both groups. Although our
hypothesis was to find decrement in microparticle levels in gruop2 there were increases and decreases in
microparticle levels in both groups (p>0,05).
Conclusion: To detect whether there is a protective effect of AMD on development of in-vitro acute GVHD
model extensive studies with multiple measurements are needed. 

Keywords

Adrenomedullin, graft versus host disease, microparticle

References

• 1) Pihusch V, Rank A, Steber R, Pihusch M, Pihusch R, Toth B, et al. Endothelial cell-derived microparticles in allogeneic hematopoietic stem cell recipients. Transplantation. 2006 27;81(10):1405-9. 2) Dickinson AM, Wang XN, Sviland L, VythDreese FA, Jackson GH, Schumacher TN, et al. In situ dissection of the graft-versus-host activities of cytotoxic T-cells specific for minor histocompatibility antigens. Nat Med 2002;8(4):410-14. 3) Woywodt A, Scheer J, Hambach L, Buchholz S, Ganser A, Haller H, et al. Circulating endothelial cells as a m a r k e r o f e n d o t h e l i a l d a m a g e i n allogeneichematopoietic stem cell transplantation. Blood 2004;103(9):3603-5. 4) Antin JH, Ferrara JL. Cytokine dysregulation and a c u t e g r a ft- v e rs u s h o st d is e a s e . Bl o o d 1992;80(12):2964-68. 5) Jimenez JJ, Jy W, Mauro LM, Soderland C, Horstman LL, Ahn YS. Endothelial cells release phenothypically and quantitavely distinct microparticles in activation and apoptosis. Thromb Res 2003;109(4):175-80. 6) Holler E, Kolb HJ, Möller A, Kempeni J, Liesenfeld S, Pechumer H, et al. Increased serum levels of tumor necrosis factor alpha precede major complications of bone marrow transplantation. Blood 1990;75(4):1011-16. 7) Isumi Y, Minamino N, Kubo A, Nishimoto N, Yoshizaki K, Yoshioka M, et al. Adrenomedullin stimulates interleukin-6 production in Swiss 3T3 cells. Biochem Biophys Res Commun 1998;244(2):325-31. 8) Chini EN, Chini CC, Bolliger C, Jougasaki M, Grande JP, Burnett JC, et al. Cytoprotective effects of adrenomedullin in glomerular cell injury: central role of cAMP signaling pathway. Kidney Int 1997:52(4):917- 25. 9) Elsasser TH, Kahl S. Adrenomedullin has multiple roles in disease stress: development and remission of the inflammatory response. Microsc Res Tech 2002;57(2):120-29. 10) Ueda S, Nishio K, Minamino N, Kubo A, Akai Y, Kangawa K, et al. Increased plasma levels of adrenomedullin in patients with systemic inflammatory response syndrome. Am J Respir Crit Care Med 1999;160(1):132-36. 11) Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins: Identification bymorphologic criteria. J Clin Invest 1973;52(11):2745-56. 12) Matsuda Y, Hara J, Osugi Y, Tokimasa S, Fujisaki H, Takai K, et al. Serum levels of soluble adhesion molecules in stem cell transplantation-related c omp li c a ti o n s. Bo n e Ma rr ow Tr a n s p l a n t. 2001;27(9):977-82. 13) Ferrara JL, Levine JE, Reddy P, Holler E. Graftversus-host disease. Lancet. 2009;373(9674):1550-61. 14) Salat C, Holler E, Kolb HJ, Pihusch R, Reinhardt B, Hiller E. Endothelial cell markers in bone marrow transplant recipients with and without acute graft-versushost disease. Bone Marrow Transplant 1997;19(9):909- 14. 15) Diamant M, Tushuizen ME, Sturk A, Nieuwland R. Cellular microparticles: new players in the field of vascular disease? Eur J Clin Invest. 2004;34(6):392-401. 16) Dursun I, Poyrazoglu HM, Gunduz Z, Ulger H, Yıkılmaz A, Dusunsel R, et al. The relationship between circulating endothelial microparticles and arterial stiffness and atherosclerosis in children with chronic kidney dis e a s e . Nephrol Di a l Tr anspl ant. 2009;24(8):2511-18. 17) González-Quintero VH, Smarkusky LP, Jiménez JJ, Mauro LM, Jy W, Hortsman LL, et al. Elevated plasma endothelial microparticles in preeclampsia. Am J Obstet Gynecol 2003;189(2):589-93. 18) Mehta JL, Szwedo J. Circulating endothelial progenitor cells, microparticles and vascular disease. J Hypertens. 2010;28(8):1611-13. Hagi-Pavli E 19) , Farthing PM, Kapas S. Stimulation of adhesion molecule expression in human endothelial cells (HUVEC) by adrenomedullin and corticotrophin. Am J Physiol Cell Physiol. 2004;286(2):239-46. 20) Sekiya M, Funahashi H, Tsukamura K, Imai T, Hayakawa A, Kiuchi T, et al. Intracellular signaling in the induction of apoptosis in a human breast cell line by water extract of Mekabu. Int J Clin Oncol 2005;10:(2)122-26. 21) Cines DB, Pollak ES, Buck CA, Loscalzo J, Zimmerman GA, McEver RP, et al. Endothelial cells in physiology and in the pathophysiology of vascular disorders. Blood 1998;91(10):3527-61. 22) Oberhammer F, Fritsch G, Schmied M, Pavelka M, Printz D, Purchio T, et al. Condensation of the chromatin at the membrane of an apoptotic nucleus is not associated with activation of an endonuclease J Cell Sci 1993;104(2):317- 26 23) Shimosawa T, Shibagaki Y, Ishibashi K, Kitamura K, Kangawa K, Kato S, et al. Adrenomedullin, an endogenous peptide, counteracts cardiovascular damage. Circulation 2002;105(1):106-11. 24) Wong LY, Cheung BM, Li YY, Tang F. Adrenomedullin is both proinflammatory and antiinflammatory: its effects on gene expression and secretion of cytokines and macrophage migration inhibitory factor in NR8383 macrophage cell line. Endocrinology 2005;146(3):1321–27. 25) Yang S, Zhou M, Fowler DE, Wang P. Mechanisms of the beneficial effect of adrenomedullin and adrenomedullin binding protein-1 in sepsis: downregulation of proinflammatory cytokines. Crit Care Med 2002;30(12):2729–35. 26) Socié G, Blazar BR. Acute graft-versus-host disease: from the bench to the bedside. Blood. 2009;114(20):4327- 36. Hamid SA 27) , Baxter GF. Adrenomedullin: regulator of systemic and cardiac homeostasis in acute myocardial infarction. Pharmacol Ther. 2005;105(2):95-112. 28) Paraskevas F. Clinical Flow Cytometri. In: Greer JP, Foester J, Rodgers GM, Paraskevas F, Glader B, Arber DA, Means RT (eds), Wintrobe's clinical Hematology (12th ed.) Lippincott Williams & Wilkins 2009, pp. 21-49. 29) Peterson RA, Krull DL, Butler L. Applications of laser scanning cytometry in immunohistochemistry and routine histopathology. Toxicol Pathol. 2008;36(1): 117–32. 30) Lewis LJ, Hoak JC, Maca RD, Fry GL. Replication of human endothelial cells in culture. Science 1973;181(4098):453-54.