Vascular endothelial growth factor (VEGF) and VEGF-receptor expression in placenta of hyperglycemic pregnant women

Term placenta, normoglycemic women. Note the immunolabeling for cytokeratin (red color) in the chorionic villi (v) in the boundary between the maternal and fetal regions indicated by the dashed line in A. In the decidual layer (D) is observed the intensely cytokeratin reactive extravillous tissue (arrowhead, A; evCt, B) and the highlight of the coating of the syncytiotrophoblast (Sc; C) in the villi chorionic. (D) Negative reaction control in which the primary antibody was replaced by non-immune serum. (fc) fetal capillary, (m) mesenchyme, (arrow) in the syncytiotrophoblast cell. The bar in the picture D indicates 400 μm in A and D; and 20 μm in B and C.

Term placentas. Portions of intermediate villi (V) in placentas of pregnant normoglycemic – N (A–D), mildly hyperglycemic – H (E–H), gestational diabetic – GD (I–L) and clinically diabetic – CD (M–P) women. Collagen fibers were stained with picrosirius red (B, F, J and N, conventional light microscopy; C, G, K and O, polarization microscopy) in the stroma of intermediate villi, mainly around vessels. Congo Red staining (D, H, L and P) was observed in vascular cells of capillaries and stromal villi (arrows) as a more intense red color. A, E, I and M – hematoxylin and eosin. The bar in A indicates 500 μm in A–E, G, M–N, P and 800 μm in F, H–L, O.

Term placentas. Portions of terminal villi in placentas of pregnant normoglycemic – N (A–C), mildly hyperglycemic – H (D–F), gestational diabetic – GD (G–I) and clinically diabetic – CD (J–L) women. Staining with picrosirius red (B, E, H and K) shows the presence of collagen in villous stroma (red) whereas staining with Congo Red highlights the presence of reactive cells (arrowhead) in placenta from women with hyperglycemia (F) and clinical diabetes (L). Reactivity in the vascular wall (arrows) was very rare (F). (arrows) villous capillaries. (*) syncytial knots. Note the large number of fetal capillaries in pregnant women with mild hyperglycemia (E). The bar in A indicates 500 μm in A, D, G and J (hematoxylin and eosin) and 250 μm in B–C, E–F, H–I and K–L.

Term placentas. VEGF expression in placentas of pregnant normoglycemic – N (A–C), mildly hyperglycemic – H (D–F), gestational diabetic – GD (G–I) and clinically diabetic – CD (J–L) women. Placental reactive components are indicated by the arrowheads. A, D, G, J and insets show regions of terminal villi whereas B, E, H and K show regions of intermediate villi. In C, F, I, L and inset show the decidua (D) with extravillous cytotrophoblast cells (EVct). (fc) fetal capillaries, (m) mesenchyme, (mv) maternal vessels, (Sc) syncytiotrophoblast, (vv) villous vessels. The bar in F indicates 200 μm in A–C and inset in C; 150 μm in D–F, J–K and inset in A and D; 400 μm in G and H, 40 μm in I and 30 μm in L.

VEGF term placental expression scores according to glycemic status of pregnant women. Strong reactions were observed in the syncytiotrophoblast cytoplasm, in muscle cells and endothelial vessels of intermediate villi, in capillary endothelial cells and in mesenchymal and cytotrophoblastic cells in all groups with exception the women with gestational diabetes.

Term placentas. VEGFR-1 expression in placentas of pregnant normoglycemic – N (A–C), mildly hyperglycemic – H (D–F), gestational diabetic – GD (G–I) and clinically diabetic – CD (J–L) women. Placental reactive components are indicated by arrows and arrowheads and are stained brown. A, D, G, J and inset show regions of terminal villi, whereas B, E, H and K represent regions of intermediate villi. In C, F, I and L note the prominent staining of extravillous cytotrophoblast cells (EVct) in the decidua (D). Staining for VEGFR-1 is present in the syncytiotrophoblast (Sc) in all groups. (EVct, arrowhead) extravillous cytotrophoblast, (m) mesenchyme, (mv) maternal vessels, (v) terminal villi, (vv) villous vessels, (*) unstained vessels. The bar in A indicates 600 μm in A, C, I–J and H; 450 μm in B, G, I and K; 120 μm in F and 150 μm in D–E and L.

VEGFR-1 term placental expression scores according to glycemic status of pregnant women. Intense reactions were observed in the normoglycemic, gestational diabetes and clinical diabetes groups but less intense in the hyperglycemic group.

Term placentas. VEGFR-2 expression in placentas of pregnant normoglycemic – N (A–C), mildly hyperglycemic – H (D–G), gestational diabetic – GD (H–J) and clinically diabetic – CD (K–M) women. Immunolabeling is seen as brown staining. Reactive cells of the extravillous trophoblast are indicated by the arrowheads. (D) basal decidua, (fc) fetal capillaries, (m) mesenchyme, (Sc) syncytiotrophoblast, (v) intermediate villus, (vv) villous capillaries. The bar in A indicates 500 μm in A–B, G–I and K; 300 μm in D–F; 250 μm in C, J, L and the inset in H and 150 μm in M.

VEGFR-2 term placental expression scores according to glycemic status of pregnant women. Intense staining was observed in the syncytiotrophoblast, capillaries, larger vessels and mesenchymal cells in of normoglycemic and hyperglycemic women. No significant reactivity was observed in clinical diabetes and gestational diabetes that was restricted to the syncytiotrophoblast cytoplasm.

Qualitative analyses of Western blotting for VEGF (A), VEGFR-1 (B) and VEGFR-2 (C) in placenta of pregnant normoglycemic (1), gestational diabetic (2), clinically diabetic (3) and mildly hyperglycemic (4) women. The membrane strips shown at the bottom of each blot in A–C correspond to membranes stained with Ponceau red after each transfer.