July 2019
Volume 60, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2019
MicroRNA-29b-3p alleviates hyperglycemia induced endothelial cell apoptosis via inhibition of DAB2IP/ASK1/JNK signaling pathway
Author Affiliations & Notes
  • Yong Zeng
    Aier school of ophthalmology, Central south university, Changsha, Hunan, China
  • Zekai Cui
    Aier eye institute, Changsha, Hunan, China
  • Jiansu Chen
    Aier school of ophthalmology, Central south university, Changsha, Hunan, China
    Aier eye institute, Changsha, Hunan, China
  • Shibo Tang
    Aier school of ophthalmology, Central south university, Changsha, Hunan, China
    Aier eye institute, Changsha, Hunan, China
  • Footnotes
    Commercial Relationships   Yong Zeng, None; Zekai Cui, None; Jiansu Chen, None; Shibo Tang, None
  • Footnotes
    Support  National Natural Scientific Fund of China 81570876
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 2655. doi:
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      Yong Zeng, Zekai Cui, Jiansu Chen, Shibo Tang; MicroRNA-29b-3p alleviates hyperglycemia induced endothelial cell apoptosis via inhibition of DAB2IP/ASK1/JNK signaling pathway. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2655.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Diabetic peripheral angiopathy is a main complication of diabetic mellitus. Recent studies have implicated microRNAs in vascular endothelium dysfunction. In this study we aim to investigate the endothelial cell protection of microRNA-29b-3p (miR-29b-3p) by targeting DAB2IP/ASK1/JNK signaling pathway in hyperglycemia condition.

Methods : Human umbilical vein endothelial cells (HUVECs) were transfected with miR-29b-3p mimic, inhibitor or its negative control. Then they exposed to 5.5 mmol/L glucose (normal glucose) and 30 mmol/L glucose (high glucose) culture media. Cell viability, wound healing and apoptosis were measured by CCK8, scratch assay and flow cytometery. MicroRNA of HUVECs was isolated and reversed into cDNA by stem-loop method. While their large RNA was extracted and reversed into cDNA by traditional method. Gene and protein expression were assayed by RT-qPCR and western blotting separately.

Results : DAB2IP and its downstream signal pathway were upregulated in HUVECs with high glucose condition, thus decreased cell viability and migration as well as promoted apoptosis (Figure 1). miR-29b-3p mimic or its inhibitor were successfully transfected into HUVECs. Upregulated miR-29b-3p in cultured cells decreased the expression of DAB2IP/ASK1/JNK as well as enhanced cell viability, migration and alleviated apoptosis. In contrast, HUVECs transfected with miR-29b-3p inhibitor increased the expression of DAB2IP/ASK1/JNK as well as decreased cell viability, migration and promoted apoptosis.

Conclusions : MiR-29b-3p enables to downregulate the expression of DAB2IP effectively. It blocks the pathway of DAB2IP/ASK1/JNK and enhances the cell viability and migration. It also alleviates the apoptosis in hyperglycemia HUVECs model. MiR-29b-3p is a potential endothelial and blood barrier protective factor.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

<span lang="EN-US" style="font-family:calibri,sans-serif; font-size:10.5pt; margin:0px"><font color="#000000">Fig.1. RT-qPCR analysis of DAB2IP/ASK1/JNK expression and cell viability assay in HUVEC with HG treatment for 48 hours. (A) (B) (C) Relative mRNA expression of DAB2IP, ASK1 AND JNK was upregulated. (D) Cell viability was decreased measured by CCK-8 assay. * p < 0.05 vs. NG.</font></span>

<span lang="EN-US" style="font-family:calibri,sans-serif; font-size:10.5pt; margin:0px"><font color="#000000">Fig.1. RT-qPCR analysis of DAB2IP/ASK1/JNK expression and cell viability assay in HUVEC with HG treatment for 48 hours. (A) (B) (C) Relative mRNA expression of DAB2IP, ASK1 AND JNK was upregulated. (D) Cell viability was decreased measured by CCK-8 assay. * p < 0.05 vs. NG.</font></span>

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