Fötal Gelişim Süresince Sığır Karaciğerindeki Sitokeratin 8 ve Sitokeratin 18 Proteinlerinin Dağılımı

Yıl 2022, Cilt: 11 Sayı: 2, 225 - 231, 30.12.2022

Uğur Topaloğlu

https://doi.org/10.31196/huvfd.1193894

Öz

Embriyonik gelişimin kontrolünde kritik rol alan sitokeratinler (CK), embriyogenezis esnasında epitel hücre gelişiminin değişen aşamalarında farklı keratinler şeklinde ekspresse edilir. CK8 ve CK18 proteinleri; çeşitli parankimatöz epitel başta olmak üzere basit epitel hücrelerinin primer keratin çifti olarak bilinmektedirler. Karaciğer; embriyonal dönemde kan yapımı ve kan hacmi regülasyonu, protein sentezi, bağışıklık sistemine katkı, büyüme sinyal yollarının endokrin kontrolü, metabolitleri depolama, safra salgısı ve detoksifikasyon gibi çok sayıda fizyolojik rollere sahiptir. Bu çalışma; CK8 ve CK18 proteinlerinin sığır fötal karaciğer hücrelerindeki dağılım ve lokalizasyonlarının immunohistokimyasal yolla belirlenmesi amacıyla yapılmıştır. Çalışmada özel kesimhanelerden temin edilen 27 adet sığır fetüsü kullanıldı. Yaşları belirlenen fetüsler; gebeliğin birinci (69-89 günlük / 9 fetüs), ikinci (99-178 / 9 fetüs) ve üçüncü (190-269 / 9 fetüs) dönemlerine ait olacak şekilde gruplandırıldı. Fetüslerden alınan karaciğer örnekleri 18 saat boyunca %10’luk formol-alkolde tespit edildi. Rutin histolojik prosedürlerden sonra elde edilen kesitlere immunohistokimyasal boyamalar yapıldı. Boyama sonucunda; CK8 ve CK18’in gebelik süresince safra kanalı epitel hücrelerinde çok güçlü seviyede ekspresse olduğu görüldü. Bununla birlikte hepatositlerde ise CK8 ve CK18’in gebelik dönemlerine göre değişen yoğunluklarda ekspresyon gösterdikleri belirlendi. Böylece CK8 ve CK18'in; sığır karaciğer gelişiminin kontrolü, hepatositlerin ve safra kanal epitel hücrelerinin bölünmesi, çoğalması ve farklılaşmaları gibi birçok role sahip olabilecekleri düşünüldü.

Anahtar Kelimeler

Fötal karaciğer, İmmunohistokimya, Sığır, Sitokeratinler, CK8, CK18, Bovine, Cytokeratins, Fetal liver, Immunohistochemistry

Teşekkür

Bu çalışmadaki bulguların değerlendirilmesinde desteklerini esirgemeyen Doç. Dr. Mehmet Erdem AKBALIK’a teşekkürlerimi sunarım.

Distribution of Cytokeratin 8 and Cytokeratin 18 Proteins in Bovine Liver During Fetal Development

Öz

Cytokeratins (CK), which play a critical role in the control of embryonic development, are expressed as different keratins at varying stages of epithelial cell development during embryogenesis. The CK8 and CK18 proteins are the primary keratin pair of simple epithelial cells, especially of various parenchymatous epithelium. The liver has many physiological roles, such as blood production and blood volume regulation, protein synthesis, contribution to the immune system, endocrine control of growth signaling pathways, metabolite storage, bile secretion, and detoxification during the embryonal period. This study was conducted to determine the distribution and localization of CK8 and CK18 proteins in bovine fetal liver cells by immunohistochemical method. In the study, 27 bovine fetuses obtained from private slaughterhouses were used. The fetuses whose ages were determined; were grouped as belonging to the first (69-89 days / 9 fetuses), second (99-178 days / 9 fetuses) and third (190-269 days/ 9 fetuses) periods of pregnancy. Then, liver tissue samples were taken from the fetuses and fixed in 10% formol-alcohol for 18 hours. The tissue samples were then subjected to routine histological procedures and immunohistochemistry staining. As a result of staining, it was observed that CK8 and CK18 were strongly expressed in bile duct epithelial cells during pregnancy. However, it has been determined that CK8 and CK18 are expressed in hepatocytes with varying densities according to the pregnancy periods. Thus, it was thought that CK8 and CK18 might have many roles, such as controlling the development of the bovine liver, division, proliferation, and differentiation of hepatocytes and bile duct epithelial cells.

Anahtar Kelimeler

Bovine, Cytokeratins, Fetal liver, Immunohistochemistry, CK8, CK18

Kaynakça

• Akiyoshi H, Inoue AM, 2012: Comparative histological study of hepatic architecture in the three orders amphibian livers. Comp Hepatol, 11 (1), 2. doi:10.1186/1476-5926-11-2.
• Basturk O, Adsay VN (2011). Immunohistology of the pancreas, biliary tract, and liver In: Diagnostic Immunohistochemistry (Third Edition), 541-592, doi.org/10.1016/B978-1-4160-5766-6.00019-4.
• Bateman AC, Hübscher SG, 2010: Cytokeratin expression as an aid to diagnosis in medical liver biopsies. Histopathology, 56, 415–425. doi: 10.1111/j.1365-2559.2009.03391.x.
• Devkota B, Sasaki M, Takahaski KI, Matsuzaki S, Matsui M, Haneda S, Takahashi M, Osawa T, Miyake YI, 2006: postnatal developmental changes in ımmunohistochemical localization of α-smooth muscle actin (SMA) and vimentin in bovine testis. J Reprod Dev, 52 (1), 43-49. doi: 10.1262/jrd.17062. Epub 2005 Nov 17.
• Eyken PV, Sciot R, Paterson A, Callea F, Kew MC, Desmet VJ, 1988: Cytokeratin expression in hepatocellular carcinoma: An immunohistochemical study 1. Hum Pathol, 19 (5), 562-568. doi: 10.1016/s0046-8177(88)80205-3.
• Feng J, Zhu R, Yin Y, Wang S, Zhou L, Lv F, Dawei Zhao D, 2021: Re-recognizing the cellular origin of the primary epithelial tumors of the liver. J Hepatocell Carcinoma, 7(8), 1537-1563. doi: 10.2147/JHC.S334935.
• Gonsebatt ME, Del Razo LM, Cerbon MA, Zuniga O, Sanchez-Pena LC, Ramirez P, 2007: Arsenite induced oxidative damage in mouse liver is associated with increased cytokeratin 18 expression. Arch Toxicol, 81, 619-626. doi:10.1007/s00204-007-0192-7.
• Guldiken N, Usachov V, Levada K, Trautwein C, Ziol M, Nahon P, Strnad P, 2014: Keratins 8 and 18 are type II acute-phase responsive genes overexpressed in human liver disease. Liver Int, 35 (4), 1203-12. http://dx.doi.org/10.1111/liv.12608. 56.
• Ham AW, Cormack DH, 1979: Histology: The pancreas, liver and gallbladder. 8th dn. JB, 694-724, Lippincott Company, Philadelphia and Toronto.
• Harris RM, Synder BG, Meyer RM, 1983: The relationship of bovine crown rump measurement to fetal age. Agri Practice, 1, 16-22.
• Jeong W, Jeong DH, Do SH, Yang HJ, Hong IH, Chang SJ, Yuan DW, Kwak DM, Kim TH, YK, Lee IS, Shik Jeong KS, 2005: Expression of cytokeratins 8 and 18 on mallory body and oval cell in rats during hepatic fibrosis. In vivo, 19, 769-776.
• Kakehashi A, Kato A, Inoue M, Ishii N, Okazaki E, Wei M, Tachibana T, Wanibuchi H, 2010: Cytokeratin 8/18 as a new marker of mouse liver preneoplastic lesions. Toxicol Appl Pharmacol, 242 (1), 47-55. doi: 10.1016/j.taap.2009.09.013.
• Korver S, Bowen J, Pearson K, Gonzalez RJ, French N, Park K, Jenkins R, Goldring C, 2021: The application of cytokeratin-18 as a biomarker for drug-induced liver injury. Archives of Toxicology, 95, 3435-3448. doi:10.1007/s00204-021-03121-0.
• Ku NO, Strnad P, Zhang BH, Tao GZ, Omary BM, 2007: Keratins let liver live: mutations predispose to liver disease and crosslinking generates Mallory-Denk bodies. Hepatology, 46 (5), 1639-1649. doi: 10.1002/hep.21976.
• Kucukoglu O, Guldiken N, Chen Y, Usachov V, El-Heliebi A, Haybaeck J, Denk H, Trautwein C, Strnad P, 2014: High-fat diet triggers Mallory–Denk body formation through misfolding and crosslinking of excess keratin 8. Hepatology, 60(1):169-78. doi: 10.1002/hep.27068.
• Kumar A, Jagannathan N, 2018: Cytokeratin: A Review on Current Concepts. Int J Orofac Biol, 2 (1), 6-11. DOI: 10.4103/ijofb.ijofb_3_18.
• Lawrence YA, Dangott LJ, Rodrigues-Hoffmann A, Steiner JM, Suchodolski JS, Lidbury JA, 2018: Proteomic analysis of liver tissue from dogs with chronic hepatitis. PLoS One, 13 (11), e0208394. doi: 10.1371/journal.pone.0208394.
• Liao J, Lowthert LA, Ku NO, Fernandez R, Omary MB, 1995: Dynamics of human keratin 18 phosphorylation: Polarized distribution of phosphorylated keratins in simple epithelial tissues. J Cell Biol, 131 (5), 1291‑301. doi: 10.1083/jcb.131.5.1291.
• Lim Y, Ku NO, 2021: Revealing the roles of keratin 8/18-associated signaling proteins involved in the development of hepatocellular carcinoma. Int. J. Mol. Sci., 22, 6401. https://doi.org/10.3390/ijms22126401.
• Michalopoulos GK, 2007: Liver regeneration. J Cell Physiol., 213 (2), 286-300. doi: 10.1002/jcp.21172.
• Moll R, Franke WW, Schiller DL, Geiger B, Krepler R, 1982: The catalog of human cytokeratins: Patterns of expression in normal epithelia, tumors and cultured cells. Cell, 31 (1), 11‑24. doi: 10.1016/0092-8674(82)90400-7.
• Naik KS, Lokanadham S, Gurushanthaiah M, 2020: Different gestational age related histogenesis of human foetal liver. Int J Anat Res, 8(1.3), 7408-11. doi: https://dx.doi.org/10.16965/ijar.2020.115.
• Omary MB, Ku NO, Toivola DM, 2002: Keratins: guardians of the liver. Hepatology, 35: (2), 251-257. doi: 10.1053/jhep.2002.31165.
• Pan X, Hobbs RP, Coulombe PA, 2013: The expanding significance of keratin intermediate filaments in normal and diseased epithelia. Curr Opin Cell Biol., 25 (1), 47-56. doi: 10.1016/j.ceb.2012.10.018.
• Schöniger-Hekele M, Petermann D, Müller C, 2006: Mutation of keratin 8 in patients with liver disease. J Gastroenterol Hepatol, 21 (9), 1466-1469. doi:10.1111/j.1440.
• Shiga A, Shirota K, 2000: Vimentin / Cytokeratin coexpression foci in a well-differentiated canine hepatocellular carcinoma. J. Vet. Med. Sci., 62 (2), 199-202. doi: 10.1292/jvms.62.199.
• Steinert PM, Marekov LN, Fraser RD, Parry DA, 1993: Keratinintermediate filament structure. Crosslinking studies yield quantitative information on molecular dimensions and mechanism of assembly. J Mol Biol, 230 (2), 436‑52. doi: 10.1006/jmbi.1993.1161
• Stosiek P, Kasper M, Karsten U, 1990: Expression of cytokeratin 19 during human liver organogenesis. Liver, 10 (1), 59-63. doi: 10.1111/j.1600-0676.1990.tb00436.x.
• Strnad P, Stumptner C, Zatloukal K, Denk H, 2008: Intermediate filament cytoskeleton of the liver in health and disease. Histochem Cell Biol, 129, 735-749. doi 10.1007/s00418-008-0431-x.
• Strnad P, Paschke S, Jang KH, Ku NO, 2012: Keratins: Markers and modulators of liver disease. Curr Opin Gastroenterol., 28 (3), 209-16. doi: 10.1097/MOG.0b013e3283525cb8.
• Subhaana CF, Talapala VR, Seethamsety S, 2020: Histogenesis of human fetal liver from 12 weeks to 36 weeks of gestation. Int J Res Med Sci., 8 (3), 931-936. doi: http://dx.doi.org/10.18203/2320-6012.ijrms20200757
• Terada T, 2017: Human ductal plate and its derivatives express antigens of cholangiocellular, hepatocellular, hepatic stellate/progenitor cell, stem cell, and neuroendocrine lineages, and proliferative antigens. Exp Biol Med (Maywood), 242 (9), 907-917. doi: 10.1177/1535370216644684.
• Toivola DM, Boor P, Alam C, Strnad P, 2015: Keratins in health and disease. Curr Opin Cell Biol., 32, 73-81. doi: 10.1016/j.ceb.2014.12.008.
• Trefts E, Gannon M, Wasserman DH, 2017: The liver. Curr Biol, 27 (21): 1147-51. doi:10.1016/j.cub.2017.09.019.1746.2006.04392.x.
• Wells MJ, Hatton MW, Hewlett B, Podor TJ, Sheffield WP, Blajchman MA, 1997: Cytokeratin 18 is expressed on the hepatocyte plasma membrane surface and interacts with thrombin-antithrombin complexes. J Biol Chem, 272 (45), 28574-81. doi: 10.1074/jbc.272.45.28574.
• Zatloukal K, Stumptner C, Fuchsbichler A, Fickert P, Lackner C, Trauner M, Denk H, 2004: The keratin cytoskeleton in liver diseases. J Pathol, 204(4), 367-376. doi: 10.1002/path.1649.