Micro-Rna335-5P Targeted Inhibition of SKLOTHO and Promoted Oxidative Stress–Mediated Aging of Endothelial Cells

Abstract

Aim: MiR-335-5p expression might induce endothelial cells (ECs) aging and target inhibit sKlotho. This study aimed to investigate whether oxidative stress evoked miR-335-5p expression and whether miR-335-5p-regulated ECs function through sKlotho. Methods: The expression of miR-335-5p was detected in human umbilical vein endothelial cells treated with H₂O₂. Subsequently, endothelial function and sKlotho expression were measured in human umbilical vein endothelial cells treated with H₂O₂ and transfected with miR-335-5p mimics or inhibitor sequences. Vector containing reporting system of sKlotho3′- untranslated region with a miR-335-5p-binding site was constructed. Results: H₂O₂ stimulation significantly increased miR-335-5p expression. Force overexpression miR-335-5p suppress ECs function and sKlotho expression. MiR-335-5p target regulated sKlotho. Conclusion: MiR-335-5p might serve as a negative factor for endothelial homeostasis and a potential treatment target for atherosclerosis.

Keywords:

• aging
• atherosclerosis
• endothelial cell
• klotho
• microRNA
• miR-335-5p
• oxidative stress
• target inhibition

Authors contributions

Y Liu carried out the studies, participated in collecting data and drafted the manuscript. P Lai, J Deng, Q Hao and H Wang performed the statistical analysis and participated in its design. B Dong helped to draft the manuscript. All authors read and approved the final manuscript.

Financial & competing interests disclosure

This study was supported by the Chengdu Science and Technology Program (2015-HM01-00417-SF), Sichuan Science and Technology Program (2018FZ0064, 2018TJPT0015 and 2018JY0094), Young researcher's initiative Program of Sichuan University (20826041A4187), 1.3.5 project for disciplines of excellence, West China Hospital, Sichuan University, National Key R&D Program of China (2017YFC0840100 and 2017YFC0840101). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

We confirm that all the research meets the ethical guidelines, including adherence to the legal requirements of the study country.

Additional information

Funding

This study was supported by the Chengdu Science and Technology Program (2015-HM01-00417-SF), Sichuan Science and Technology Program (2018FZ0064, 2018TJPT0015 and 2018JY0094), Young researcher's initiative Program of Sichuan University (20826041A4187), 1.3.5 project for disciplines of excellence, West China Hospital, Sichuan University, National Key R&D Program of China (2017YFC0840100 and 2017YFC0840101). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.