Selda İldan ÇALIM, Oya SAYIN, Seren Gülşen GÜRGEN, Ayşe Tuç YÜCEL, Ferihan ÇETİN

Sıçanlarda Gebelik Süresince Yeşil Çay Tüketiminin Maternal ve Neonatal Hepatositlerde Sitokeratin-18 Üzerindeki Etkisi

Amaç: Bu çalışmada, gebelikleri süresince yeşil çay ekstraktıyla beslenen sıçanların ve yavrularının karaciğer dokularında sSitokeratin-18 (SK-18) düzey ve ekspresyonunu değerlendirmek amaçlanmıştır. Yöntem: On sekiz adet Wistar albino gebe sıçan, iki gruba ayrıldı: kontrol grubu ve (oral gavaj ile 50 mg/kg yeşil çay ekstraktı verilenuygulanan) yeşil çay grubu. Yirmi bir günlük gebelikten sonra, her iki gruptaki anne sıçanların ve doğdukları ilk gün yavruların karaciğer dokuları çıkarıldı. Bu doku örneklerinde SK-18 ekspresyonu ve düzeyi immünohistokimyasal olarak ve enzime bağlı immünosorbent analiz (ELISA) ile değerlendirildi. Bulgular: İki grupta da maternal dokularda santral venlerin çevresindeki hepatositlerin hücre zarları yakınında kuvvetli SK-18 immünoreaksiyonu gözlendi. Kontrol grubu yenidoğan dokularında santral ven çevresindeki hepatositlerde zayıf SK-18 immünoreaksiyonu gözlenirken, yeşil çay grubunda hepatositlerin hücre zarı yakınında oldukça kuvvetli SK-18 immünoreaksiyonu gözlendi. Biyokimyasal incelemede de, maternal SK-18 düzeyleri her iki grupta da yüksek olup birbirinden istatistiksel olarak anlamlı farklılık göstermezken, neonatal SK-18 düzeyleri kontrol grubuna kıyasla yeşil çay grubunda anlamlı biçimde daha yüksekti. Sonuç: İmmünohistokimya ve ELISA sonuçlarımız gebelik süresince maternal yeşil çay tüketiminin yenidoğan karaciğerinde hücre hasarına neden olabileceğini düşündürmektedir.

The Effects of Green Tea Consumption during Pregnancy on the Cytokeratin-18 in Maternal and Neonatal Hepatocytes in Rats

Aim: In this study, we aimed to evaluate the cCytokeratin-18 (CK-18) levels and expression in liver tissues of rats that were fed with green tea extract during pregnancy and their pups. Methods: Eighteen pregnant Wistar albino rats were divided into two groups: the control group and the green tea group (orally gavaged with 50 mg/kg of green tea extract). After 21- days of gestation, liver tissues were removed from the mother rats and their pups on the first postnatal day in both groups. The CK-18 levels and expression in these tissue samples were evaluated immunohistochemically and by use of enzyme-linked immunosorbent assay (ELISA). Results: Strong CK-18 immunoreaction was observed near the cell membranes of hepatocytes around the central veins in maternal liver tissues in both groups. While a weak CK-18 immunoreaction was observed in hepatocytes around the central veins in the control group neonatal tissues, a strong CK-18 immunoreaction was observed near the cell membranes of hepatocytes in the green tea group. Biochemically also, while maternal CK-18 levels were high in both groups with no statistically significant difference, neonatal CK-18 levels were significantly higher in the green tea group than in the control group. Conclusion: Our immunohistochemical and ELISA results suggest that maternal consumption of green tea during pregnancy may lead to cell injury in the neonatal liver.

PDF

___

1. Singh BN, Shankar S, Srivastava RK. Green tea catechin, epigallocatechin-3-gallate (EGCG): mechanisms, perspectives and clinical applications. Biochem Pharmacol. 2011;82(12):1807–21.
2. Bun SS, Bun H, Guédon D, Rosier C, Ollivier E. Effect of green tea extracts on liver functions in Wistar rats. Food Chem Toxicol. 2006;44(7):1108–13.
3. Nakamoto K, Takayama F, Mankura M, Hidaka Y, Egashira T, Ogino T, ve ark. Beneficial effects of fermented green tea extract in a rat model of non-alcoholic steatohepatitis. J Clin Biochem Nutr. 2009;44(3):239–46.
4. Zhi Z, Froh M, Connor HD, Li X, Conzelmann LO, Mason RP, ve ark. Prevention of hepatic ischemia-reperfusion injury by green tea extract. Am J Physiol Gastrointest Liver Physiol. 2002;283:957–64.
5. Zhong Z, Froh M, Lehnert M, Schoonhoven R, Yang L, Lind H, ve ark. Polyphenols from Camellia sinenesis attenuate experimental cholestasis induced liver fibrosis in rats. Am J Physiol Gastrointest Liver Physiol. 2003;285:1004–13.
6. Li YM, Zhang XG, Zhou HL, Chen SH, Zhang Y, Yu CH. Effects of tea polyphenols on hepatic fibrosis in rats with alcoholic liver disease. Hepatobiliary Pancreat Dis Int. 2004;3:577–9.
7. Arteel GE, Uesugi T, Bevan LN, Gäbele E, Wheeler MD, McKim SE, ve ark. Green tea extract protects against early alcohol-induced liver injury in rats. Biol Chem. 2002;383(3– 4):663–70.
8. Chen JH, Tipoe GL, Liong EC, So HSH, Leung KM, Tom WM, ve ark. Green tea polyphenols prevent toxin-induced hepatotoxicity in mice by down-regulating inducible nitric oxide-derived prooxidants. Am J Clin Nutr. 2004;80:742–51.
9. Oz HS, McClain CJ, Nagasawa HT, Ray MB, Villiers WJ, Chen TS. Diverse antioxidants protect against acetaminophen hepatotoxicity. J Biochem Mol Toxicol. 2004;18:361–8.
10. Mazzanti G, Menniti-Ippolito F, Moro PA, Cassetti F, Raschetti R, Santuccio C, ve ark. Hepatotoxicity from green tea: a review of the literature and two unpublished cases. Eur J Clin Pharmacol. 2009;65:331–41.
11. Lambert JD, Kennett MJ, Sang S, Reuhl KR, Ju J, Yang CS. Hepatotoxicity of high oral dose (-)-epigallocatechin-3- gallate in mice. Food Chem Toxicol. 2010;48(1):409–16.
12. Galati G, Lin A, Sultan AM, O’Brien PJ. Cellular and in vivo hepatotoxicity caused by green tea phenolic acids and catechins. Free Radic Biol Med. 2006;15:40(4):570–80.
13. Chow HHS, Cai Y, Hakim IA, Crowell JA, Shahi F, Brooks CA, ve ark. Pharmacokinetics and safety of green tea polyphenols after multiple-dose administration of epigallocatechin gallate and polyphenon E in healthy individuals. Clin Cancer Res. 2003;9(9):3312–9.
14. Chengelis CP, Kirkpatrick JB, Regan KS, Radovsky AE, Beck MJ, Morita O, ve ark. 28-day oral gavage toxicity studies of green tea catechins prepared for beverages in rats. Food Chem Toxicol. 2008;46(3):978–89.
15. Morita O, Kirkpatrick JB, Tamaki Y, Chengelis CP, Beck MJ, Bruner RH. Safety assessment of heat-sterilized green tea catechin preparation: a 6-month repeat-dose study in rats. Food Chem Toxicol. 2009;47(8):1760–70.
16. Wang CC, Chu KO, Chong WS, Li WY, Pang CP, Shum ASW, ve ark. Tea epigallocatechin-3-gallate increases 8-isoprostane level and induces caudal regression in developing rat embryos. Free Radic Biol Med. 2007;43(4):519–27.
17. Correa A, Stolley A, Liu Y. Prenatal tea consumption and risks of anencephaly and spina bifida. Ann Epidemiol. 2000;10(7):476–7.
18. Ismail SA, El-Saadany S, Ziada DH, Zakaria SS, Mayah WW, Elashry H, ve ark. Cytokeratin-18 in diagnosis of HCC in patients with liver cirrhosis. Asian Pac J Cancer Prev. 2017;18(4):1105–11.
19. Yilmaz Y. Systematic review: caspase-cleaved fragments of cytokeratin 18—the promises and challenges of a biomarker for chronic liver disease. Aliment Pharmacol Ther. 2009;30(11–12):1103–9.
20. Abdel-Majeed S, Mohammad A, Shaima AB, Mohammad R, Mousa SA. Inhibition property of green tea extract in relation to reserpine-induced ribosomal strips of rough endoplasmic reticulum (rER) of the rat kidney proximal tubule cells. J Toxicol Sci. 2009;34(6):637–45.
21. Gürgen SG, Sayın O, Cetin F, Tuç Yücel A. Transcutaneous electrical nerve stimulation (TENS) accelerates cutaneous wound healing and inhibits pro-inflammatory cytokines. Inflammation. 2014;37(3):775–84.
22. Gürgen SG, Karakuş AÇ, Çeçen D, Özen G, Koçtürk S. Usage of whey protein may cause liver damage via inflammatory and apoptotic responses. Hum Exp Toxicol. 2015;34(7):769–79.
23. Morita O, Knapp JF, Tamaki Y, Stump DG, Moore JS, Nemec MD. Effects of green tea catechin on embryo/fetal development in rats. Food Chem Toxicol. 2009;47(6):1296–303.
24. Chu KO, Wang CC, Chu CY, Chan KP, Rogers MS, Choy KW, ve ark. Pharmacokinetic studies of green tea catechins in maternal plasma and fetuses in rats. J Pharm Sci. 2006;95:1372–81.
25. Chu KO, Wang CC, Chu CY, Choy KW, Pang CP, Rogers MS. Uptake and distribution of catechins in fetal organs following in utero exposure in rats. Hum Reprod. 2006;22:280–7.
26. Isbrucker RA, Edwards JA, Wolz E, Davidovich A, Bausch J. Safety studies on epigallocatechin gallate (EGCG) preparations. Part 3: teratogenicity and reproductive toxicity studies in rats. Food Chem Toxicol. 2006;44(5):6516–61.