Preliminary Research of Main Components of Dll4/ Notch-VEGF Signaling Pathway Under High-Glucose Stimulation in vitro

Abstract

Purpose

To establish a high-glucose (HG) stressed cell model and study the expression of main components of the Dll4/Notch-VEGF signaling pathway under high-glucose stimulation.

Methods

A model of HG-conditioned cells (human umbilical vein endothelial cells, HUVECs) was first established, and then the expression of Dll4, Notch1, Notch4 and VEGF in HG-stressed cells with or without Notch pathway blockage was analyzed by RT-PCR and Western blot. To observe cell migration, we also evaluated the Transwell assay.

Results

HUVECs stimulated with 30mmol/L HG was selected as a cell model. RT-PCR and Western blot results showed that HG stimulation induced the expression of Dll4, Notch1 and VEGF and downregulated Notch4. The expressions were reversed after Notch pathway blockage; meanwhile, the blockage of Notch pathway inhibited cell migration under HG condition.

Conclusion

The function of Notch4 in responses to HG stimulation deserves further researching. Combination therapy by blocking Dll4/Notch and VEGF pathways may provide us with a new way for anti-neovascularization.

Keywords:

• Dll4
• Notch1
• Notch4
• VEGF
• endothelial cells

Acknowledgments

We thank the ophthalmic laboratory of Chengdu University of Traditional Chinese Medicine for technical assistance. Na Gao and Linghui Xiao are co-first authors for this study.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This work was supported by the research program of Health and Family Planning Commission of Sichuan Province, China (17PJ534).