RETİNOİK ASİT UYGULANAN SIÇANLARIN PLASENTALARINDA MMP-2 VE MMP-13 İMMÜNREAKTİVİTESİNİN İNCELENMESİ

Year 2010, Volume: 73 Issue: 2, 46 - 51, 10.11.2011

Tuğba Kotil Leyla Tapul

Abstract

Amaç: Matriks metalloproteinazlar (MMP) hücrelerarası matriks yıkımında rol alarak implantasyonun gerçekleşmesine yardımcı olan enzimlerdir. Retinoik asit (RA); MMP üretimi üzerine etkisi olduğu düşünülen, trofoblast invazyonunda rolü olan bir moleküldür. Çalışmamızda retinoik asit uygulanan gebe sıçanların plasentalarında MMP-2 ve MMP-13 immünreaktivitesinin incelenmesi amaçlandı. Gereç ve Yöntem: Çalışmada kontrol (8 adet) ve deney (8 adet) gruplarında toplam 16 adet dişi Spraque Dawley türü sıçan kullanıldı. Sıçanlar gebe bırakılarak deney grubundakilere gastrulasyon döneminde 3 gün süre ile 40mg/kg/gün 13-cis retinoik asit oral yolla uygulandı. Gebeliğin orta döneminde (10-14 gün) plasentalardan alınan kesitler immünhistokimyasal olarak Streptavidin- Biyotin yöntemi kullanılarak rabbit poliklonal MMP-13 antikoru ve mouse monoklonal MMP-2 antikoru ile işaretlendi. Bulgular: Kontrol grubuna ait plasentalarda MMP-2 immünreaktivitesi trofoblastik dev hücrelerde kuvvetli pozitif (+++), koriyo-amniyotik zarda ise orta derecede pozitif (++) bulundu. RA uygulanan grupta trofoblastik dev hücrelerde MMP-2 immünreaktivitesi saptanmadı, buna karşın koriyo-amniotik zarda kuvvetli pozitiflik mevcuttu. MMP-13 immünreaktivitesi ise kontrol grubu dev hücrelerinde ve korio-amniyotik zarda orta derecede (++) pozitifti. RA grubunda dev hücrelerde değişiklik görülmezken koriyo-amniyotik zarda kuvvetli pozitif (+++) işaretlenme mevcuttu. Sonuç: Eksojen olarak uygulanan retinoik asitin trofoblastik dev hücrelerde ve desiduada MMP-2 sentezini baskılayarak implantasyonu olumsuz etkileyebileceği ve koriyo-amniyotik zarda ise MMP-2 ve MMP-13‟ü arttırmak suretiyle membran stabilizasyonunu bozabileceği, bunlara bağlı olarak hayatta kalımı engelleyebileceği kanaatine varıldı.

Keywords

Plasenta, retinoik asit, matriks metalloproteinaz-2, matriks metalloproteinaz-13

References

• Alexander CM, Hansell EJ, Behrendtsen O, Flannery ML, Kishnani N, Hawkes SP, Werb Z. Expression and function of matriks metalloproteinases and their inhibitors at the maternal-embryonic boundary during mouse embryo implantation. Development 1996; 122: 1723-1736.
• Braunhut SJ, Moses MA. Retinoids modulate endothelial cell production of matrix-degrading proteases and tissue inhibitors of metalloproteinases (TIMP). The Journal of Biological Chemistry 1994; 269 : 13472-13479.
• Das SK, Yano S, Wang J, Edwards DR, Nagase H, Dey SK. Expression of Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases in the Mouse Uterus During the Peri-Implantation Period. Developmental Genetics 1997; 21:44-54.
• El-Hashash AH, Esbrit P, Kimber SJ. PTHrP promotes murine secondary trophoblast giant cell differentiation through induction of endocycle, upregulation of giant- cell-promoting transcription factors and suppression of other trophoblast cell types. Differentiation 2005; 73: 154–174.
• Fortunato SJ, Menon R. Screening of Novel Matrix Metalloproteinases (MMPs) in Human Fetal Membranes. Journal of Assisted Reproduction and Genetics 2002; 19:483-486.
• Goldman S, Weiss A, Eyali V, Shalev E. Differential activity of gelatinases (matrix metalloproteinases 2 and 9) in the fetal membranes and decidua, associated with labour. Molecular Human Reproduction 2003; 9: 367- 373.
• Hasan S, Nakajima M. Retinoic acid synergizes with cyclic AMP to enhance MMP-2 basal promoter activity. Biochemical and Biophysical Research Communications 1999; 258: 663-667.
• Hendrix MJC, Wood WR, Seftor EA, Lotan D, Nakajima M, Misiorowski RL, SeftorREB, Stetler-Stevenson WG, Bevacqua SJ, Liotta LA, Sobel ME, Lotan R. Retinoic acid inhibition of human melanoma cell invasion through a reconstituted basement membrane and its relation to decreases in the expression of proteolytic enzymes and motility factor receptor. Cancer Res 1990; 50:4121-4130.
• Huang F. Effects of retinoic acid on morula-stage embryo development in mice. Chang Gung Med J 2008;31:44-51.
• Kanai-Azuma M, Kanai Y, Matsuda H, Kurohmaru M, Tachi C, Yazaki K, Hayashi Y. Nerve growth factor 50
• Teesalu T, Masson R, Basset P, Blasi F, Talarıco D. Expression of matriks metalloproteinases during murine chorioallantoic Dynamics 1999; 214: 248-258. maturation. Developmental
• Tremblay GB, Kunath T, Bergeron D, Lapointe L, Champigny C, Bader JA, Rossant J, Giguere V. Diethylstilbestrol differentiation as a ligand of orphan nuclear receptor ERR beta. Genes Dev 2001; 15: 833–838. stem cell
• Uchide K, Ueno H, Inoue M, Sakaı A, Fujimoto N, Okada Y. Matrix metalloproteinase-9 and tensile strength of fetal membranes in uncomplicated labor. Obstet Gynecol 2000; 95: 851-855.
• Ulug U, Goldman S, Ben-Shlomo I, Shalev E. Matrix metalloproteinase (MMP)- 2 and MMP-9 and their inhibitor, TIMP-1, in human term decidua and fetal membranes: the effect of prostaglandin F2α and indomethacin. Molecular Human Reproduction 2001; 7: 1187-1193.
• Walter I, Schönkypl S. Extracellular matrix components and matrix degrading enzymes in the feline placenta during gestation. Placenta 2006; 27: 291-306.
• Weiss A, Goldman S, Shlomo IB, Eyali V, Leibovitz S, Shalev E. Mechanisms of matrix metalloproteinase-9 and matrix metalloproteinase-2 inhibition by N-acetylcysteine in the human term decidua and fetal membranes. Am J Obstet Gynecol 2003; 189:1758-1763.
• Winterhager E, Reuss B, Hellmann P, Spray DC, Gruemmer R. Gap junctionand tissue invasion: A comparison of tumorigenesis and pregnancy. Clin. Exp. Pharmacol. Physiol 1996; 23: 1058-1061.
• Xu P, Alfaidy N, Challis JRG. Expression of matriks metalloproteinase (MMP)-2 and MMP-9 in human placenta and fetal membranes in relation to preterm and term labor. The Journal of Clinical Endocrinology & Metabolism 2002; 87: 1353-1361.
• Xu P, Wang Y, Zhu S, Luo S, Piao Y, Zhuang K. Expression of matrix metalloproteinase-2, -9, and -14, Tissue Inhibitors of metalloproteinase-1, and Matrix proteins in human placenta during the first trimester. Biology of Reproduction 2000; 62: 988-994.
• Yan J, Tanaka S, Oda M, Makino T, Ohgane J, Shiota K. Retinoic acid promotes differentiation of trophoblast stem cells to a giant cell fate. Developmental Biology 2001; 235: 422-432.
• Expression and importance of matrix metalloproteinase 2
• and 9 (MMP-2 and -9) in human trophoblast invasion.
• Reproductive Biology and Endocrinology 2004; 2:59.
• Stephanou A, Handwerger S. Retinoic acid and thyroid hormone regulate placental lactogen expression in human trophoblast cells. Endocrinology 1995; 136:933-938.