Publicación: UMBILICAL CORD PLASMA FROM PREECLAMPTIC PREGNANCIES REDUCE BRAIN ENDOTHELIAL CELL MIGRATION
Fecha
2022
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OBJECTIVES: ANALYZE WHETHER UMBILICAL CORD PLASMA FROM PREECLAMPTIC PREGNANCIES (U-PE) REDUCES BRAIN ENDOTHELIAL CELL MIGRATION AS AN UNDERLYING MECHANISM OF IMPAIRED BRAIN ANGIOGENESIS.
METHODS: HUMAN CEREBRAL MICROVASCULAR ENDOTHELIAL CELLS (HCMEC/D3) WERE INCUBATED (12 H, 1% V/V) WITH UMBILICAL CORD PLASMAS FROM NORMOTENSIVE (N=20, U-NP) OR WOMEN WITH PREECLAMPSIA (N=20, U-PE). CELL VIABILITY (MTT ASSAY), CELL PROLIFERATION (DETECTION OF BROMOURIDINE), AND CELL MIGRATION (WOUND HEALING IN VITRO ASSAY) WERE MEASURED. SYNTHESIS AND RELEASE OF VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) AND THE PRESENCE OF ITS RECEPTOR TYPE 2 IN THE INACTIVE (VEGFR2) OR ACTIVE PRO-MIGRATORY PHOSPHORYLATION (Y951-VEGFR2) WERE ANALYZED BY WESTERN BLOT. THE SOLUBLE VEGF RECEPTOR TYPE 1 (OR SFLT-1) WAS MEASURED USING ELISA. ALSO, F-ACTIN POLYMERIZATION/DEPOLYMERIZATION PROTEINS, INCLUDING COFILIN, PHOSPHO-COFILIN, AND ARP2/ARP3 COMPLEX, WERE ANALYZED BY WESTERN BLOT. THE LENGTH AND WIDTH OF F-ACTIN FIBERS (F-ACTIN) WERE EVALUATED USING PHALLOIDIN FLUORESCENCE
RESULTS: COMPARED TO PLASMA FROM U-NP, U-PE SIGNIFICANTLY REDUCED CELL PROLIFERATION (P=0.04) AND CELL MIGRATION (P=0.02) WITHOUT AFFECTING CELL VIABILITY. A REDUCTION IN THE VEGFR2 PROTEIN AMOUNT IN BOTH THE INACTIVE AND Y951 PHOSPHORYLATED ISOFORM (PARTICULARLY IN THE CYTOPLASM) WAS FOUND IN CELLS EXPOSED TO U-PE. THIS EFFECT OF U-PE WAS ASSOCIATED WITH REDUCED COFILIN EXPRESSION AND IN THE WIDTH OF F-ACTIN FIBERS, PARTICULARLY IN THE MIDDLE AND TIP SECTIONS OF THE CELLS. IN CONTRAST, NO SIGNIFICANT CHANGES WERE OBSERVED IN THE SYNTHESIS/RELEASE OF VEGF, NEITHER IN THE PLASMA LEVELS NOR THE RELEASE OF SFLT-1. ALSO, NO CHANGES IN PHOSPHO-COFILIN AND ARP2/ARP3 COMPLEX WERE FOUND IN CELLS EXPOSED TO U-PE.
CONCLUSION: U-PE LEADS TO REDUCED BRAIN ENDOTHELIAL CELL MIGRATION ASSOCIATED WITH DOWNREGULATION OF THE PRO-MIGRATORY PHOSPHORYLATION OF VEGFR2 AND COFILIN LEVELS THAT MAY LEAD TO HARMED POLYMERIZATION OF F-ACTIN. THESE FINDINGS
