Expression of ADAMTS13 and PCNA in the placentas of gestational diabetic mothers / Expresión de ADAMTS13 y PCNA en las placentas de madres diabéticas gestacionales

SUMMARY: GDM is linked with overexpression of inflammatory cytokines and increased oxidative stress, leading to endothelial dysfunction and vascular disorder. Weaimed to examine the expression of ADAMTS13 and PCNA in the placentas of gestational diabetes mellitus (GDM) patients to investigate the effects of hypoxia, induced by GDM, on proliferation and extracellular matrix formation in the maternal and fetal placenta cells. A total of 60 placentas were collected from pregnant women admitted to the obstetrics clinic. Thirty of them were diagnosed with GDM, and 30 of them were diagnosed with non-GDM patients. Samples were fixed in 10 % formaldehyde, after routine follow-up, embedded in paraffin wax. Sections of 5 µm were cut stained with Mayer Hematoxylin-Eosin, examined under a light microscope. Sections for immunohistochemical analysis were cut and processed for antigen retrievalin citrate solution. Sections were incubated with ADAMTS13 and PCNA primary antibodies, counterstained with hematoxylin, and evaluate under a light microscope. In histopathological examination, the non-diabetic placentas showed that decidua cells in the maternal region were polygonal with oval nuclei and organized in groups. In the GDM group, there were pyknosis and apoptotic changes in decidua cell nuclei. Vacuolar areas were observed in large cavities in maternal connective tissue. Inflammation and dilatation with congestion were observed in the blood vessels of the villus. In the GDM group, positive ADAMTS13 expression was observed in the decidua cells vascular endothelial cells, and surrounding connective tissue fibroblast cells. In the GDM group, a significant increase in PCNA expression was observed in decidua cells, connective tissue cells and endothelial cells. Functional changes in ADAMTS13 proteases and PCNA were thought to induce maternal and fetal complications by stimulating extracellular matrix development.

RESUMEN: La diabetes gestacional está relacionada con la sobreexpresión de citocinas inflamatorias y aumento del estrés oxidativo, lo que lleva a una disfunción endotelial y un trastorno vascular. Nuestro objetivo fue examinar la expresión de ADAMTS13 y PCNA en las placentas con diabetes mellitus gestacional (DMG) para investigar los efectos de la hipoxia inducida por DMG sobre la proliferación y formación de matriz extracelular en células placentarias maternas y fetales. Se recolectaron un total de 60 placentas de mujeres embarazadas ingresadas a la consulta de obstetricia. Treinta de ellas fueron diagnosticadas con DMG y 30 diagnosticadas sin DMG. Las muestras se fijaron en formaldehído al 10 %, y luego de un seguimiento de rutina, fueron embebidas en parafina. Se cortaron secciones de 5 µm teñidas con hematoxilina-eosina de Mayer, las que fueron examinadas bajo un microscopio óptico. Se cortaron y procesaron las secciones para el análisis inmunohistoquímico para la recuperación de antígeno en solución de citrato. Las secciones se incubaron con anticuerpos primarios ADAMTS13 y PCNA, se contratiñeron con hematoxilina y se evalua- ron con un microscopio óptico. En el examen histopatológico, las placentas no diabéticas mostraron que las células de la decidua en la región materna eran poligonales con núcleos ovalados y organizadas en grupos. En el grupo de DMG, se observó picnosis y cambios apoptóticos en los núcleos de las células de la decidua. Se observaron áreas vacuolares en el tejido conectivo materno. En los vasos sanguíneos de las vellosidades se observó inflamación y dilatación con congestión. En el grupo de DMG, se observó expresión positiva de ADAMTS13 en las células de la decidua, en las células endoteliales vasculares y en los fibroblastos del tejido conectivo circundante. En el grupo de DMG se observó un aumento significativo de la expresión de PCNA en células de la decidua, células de tejido conectivo y en las células endoteliales. Se considera que los cambios funcionales en las proteasas ADAMTS13 y PCNA inducen a complicaciones maternas y fetales al estimular el desarrollo de la matriz extracelular.

Effect of Vitamin D Status on Von Willebrand Factor and ADAMTS13 in Diabetic Patients on Chronic Hemodialysis

Von Willebrand factor (vWF) is a glycoprotein with a crucial role in the formation of platelet thrombi, and ADAMTS13 is the main enzyme responsible for vWF cleavage. Both are important in the relationship between diabetic nephropathy, hypercoagulability, and cardiovascular disease. This study evaluated a potential relationship between vitamin D (vitD) levels, vWF, ADAMTS13 activity, and inflammation in diabetic patients on chronic hemodialysis (HD). Blood samples from 52 diabetic patients on chronic HD were obtained to determine vitD levels, vWF, and ADAMTS13 activity, and inflammatory markers. HD patients were grouped according to 25-hydroxyvitamin D [25(OH) VitD]25 nmol/L (n=36). vWF antigen and vWF activity were elevated in both groups, with an average of 214.3±82.6% and 175.8±72.6%, respectively. Average ADAMTS13 activity was within the normal range in both groups. Blood samples from the vitD <25 nmol/L group showed a positive correlation between c-reactive protein (CRP) and vWF levels (P=0.023; r=0.564; 95% confidence interval=0.095-0.828), with a negative correlation between HbA1c and 25(OH) VitD (P=0.015; r=-0.337; 95% confidence interval=-0.337-0.19). Diabetic patients on chronic HD had elevated vWF levels and activity with no significant change in ADAMTS13 activity. The correlation between CRP and vWF levels in the 25(OH) VitD<25 nmol/L group suggests inflammatory-related endothelial dysfunction in these patients.