The Role Of Notch1 Signaling Pathways Involved In The Angiogenesis Of Human Placenta

Part One To investigate the change of Notch1 Signaling in Hypoxic HUVECs【 Objective 】 To detect the angiogenic ability of HUVECs and the expression of Notch1/Dell4/Jagged1 m RNA and protein in hypoxic groups and normal groups.【Methods】1. The Matrigel network formation assay was used to observe the angiogenic ability of HUVECs in hypoxic groups and the normal groups as control.2. The Real-Time PCR was used to detect the expression of Notch1/Dell4/Jagged1 m RNA in HUVECs in hypoxic groups and the normal groups as control.3. The Western Blot was applied to detect the expression of Notch1/Dell4/Jagged1 in protein level in HUVECs in hypoxic groups and the normal groups as control.【Result】1. The angiogenic ability of HUVECs was seriously reduced in hypoxic groups as compared with control groups. Mainly show as: the broken blood vessels grow in quantity and the integrity of the vascular lumen reduced;2. The relative expressions of Notch1/Dell4/Jagged1 m RNA were obviously reduced in hypoxic groups as compared with control groups(P<0.05).But the expressions of them showed no difference in each time point(P>0.05);3. The relative expressions of Notch1/Dell4 in protein level were obviously reduced in hypoxic groups as a compared with control groups with the extending of time(P<0.05).And the expression of Jagged1 in protein level also significantly reduced in hypoxic groups(P<0.05). But the expression of jagged1 showed no difference in each time point(P>0.05)【Conclusion】 The reduced angiogenic ability of HUVECs and the decreased expressions of Notch1/Dell4/Jagged1 in hypoxic HUVECs suggesting the Notch1 signaling may play an important role in the functional regulation of HUVECs. Maybe the imbalance expression of notch1 signaling will contribute to the angiogenesis of human placenta.Part Two The Effect of Notch1 Signaling on migration capability of HUVECs.【Objective 】To investigate the effect of notch signaling on the migration capability of HUVECs.【Methods】The cell wound scratch assay was used to detect the migration capability of HUVECs after DAPT treatment and Jag1 peptidetreatment.【Result】1. The migration capability of HUVECs was significantly inhibited after treatment with DAPT, an inhibitor of r-secretase, which can effectively block the Notch signaling;2. The migration capability of HUVECs was promoted after treatment with JAG-1, a peptide fragment of Jagged-1, which can be used as an agonist to activate the Notch pathway.【Conclusion】DAPT can significantly inhibit the migration capability of HUVECs, while JAG-1 can promote the migration capability of HUVECs. All these suggest that the Notch1 signaling pathways may play an important role in the functional regulation of HUVECs,which may contribute to the angiogenesis of human placenta.