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Ablation of NF-κB expression by small interference RNA prevents the dysfunction of human umbilical vein endothelial cells induced by high glucose

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Abstract

Diabetes is a major independent risk factor for cardiovascular disease and stroke. High glucose (HG) reduces endothelial cell (EC) proliferation with a concomitant increase in apoptosis. HG also induces the translocation of nuclear factor (NF)-κB in human umbilical vein endothelial cells (HUVECs). However, data regarding the relationship between NF-κB signaling and HG-induced endothelial dysfunction are limited. In the present study, we constructed an NF-κB-targeting RNA interference (RNAi) adenovirus vector and cultured HUVECs in 5.5, 20.5, or 30.5 mM d-glucose or in daily alternating 5.5 or 30.5 mM d-glucose. We assessed the effects of the NF-κB pathway on proliferation under HG conditions by measuring bromodeoxyuridine incorporation and conducting methyl thiazolyltetrazolium assays. We also tested apoptosis by performing flow cytometry and terminal deoxynucleotidyl transferase nick-end labeling assay. The RNAi adenovirus effectively downregulated expression of the p65 protein in HUVECs for more than 6 days. Blockage of the NF-κB pathway with the RNAi adenovirus substantially protected HUVECs from decreased proliferation and reduced cellular apoptosis in HG conditions. These findings may explain how hyperglycemia promotes dysfunction of ECs and could elucidate a potential new target for therapeutic interventions.

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Acknowledgments

This work was supported by Grant C071002 for natural science foundation from Fujian province of China and grant from the Ministry of Health of the People’s Republic of China (WKJ 2005-2-021).

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Authors and Affiliations

  1. Department of Endocrinology, Fujian Provincial Hospital, Fujian Medical University, 92 Huqian Road, Box 4-704, Fuzhou, Fujian, China

    Gang Chen, Xiaoyan Shen, Jin Yao, Feng Chen, Yufang Qiao, Tingting You, Fenghui Lin, Xiaowen Fang, Xin Zou & Lixiang Lin

  2. Department of Molecular Medicine Research Center, Fujian Medical University, Fuzhou, China

    Xu Lin

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  1. Gang Chen
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  2. Xiaoyan Shen
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  3. Jin Yao
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Correspondence to Gang Chen.

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This study had been approved by IRB of Fujian Provincial Hospital and all patients had signed the consent form.

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Chen, G., Shen, X., Yao, J. et al. Ablation of NF-κB expression by small interference RNA prevents the dysfunction of human umbilical vein endothelial cells induced by high glucose. Endocr 35, 63–74 (2009). https://doi.org/10.1007/s12020-008-9120-8

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  • DOI: https://doi.org/10.1007/s12020-008-9120-8

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