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miR-30e-5p Regulates Autophagy and Apoptosis by Targeting Beclin1 Involved in Contrast-induced Acute Kidney Injury

Author(s):

Xiaoqin Liu, Qingzhao Li , Lixin Sun, Limei Chen, Yue Li, Beibei Huang, Yunshuang Liu and Chunyang Jiang *   Pages 1 - 11 ( 11 )

Abstract:


Aims: This study aims to verify if miR-30e-5p targets Beclin1 (BECN1), a key regulator of autophagy, and investigate the function of miR-30e-5p and Beclin1 through mediating autophagy and apoptosis in contrast-induced acute kidney injury (CIAKI).

Methods: Human renal tubular epithelial HK-2 cells were treated with Urografin to construct a cell model of CI-AKI. Real-time reverse transcription-polymerase chain reaction was used to detect gene expression. The dual-luciferase reporting assay and endogenous validation were used to verify targeting and regulating function. The expressions of protein were detected using Western blot. Cell proliferation was detected using methylthiazolyldiphenyl- tetrazolium bromide (MTT) assay. Cell apoptosis was detected using terminal- deoxynucleoitidyl transferase mediated nick end labeling assay, and autophagy was detected using transmission electron microscopy.

Results: HK-2 cells exposed to Urografin for 2 h induced a significant increase in miR-30e-5p. miR-30e-5p had a targeting effect on Beclin1. Moreover, Urografin exposure can enhance cell apoptosis by increasing caspase 3 gene expression and inhibiting autophagy, which was induced by decreased Beclin1 expression regulated by miR-30e-5p, thereby resulting in renal cell injury. Downregulation of miR-30e-5p or upregulation of Beclin1 restored cell vitality by promoting autophagy and suppressing apoptosis in Urografin-treated cells.

Conclusion : Urografin increased the expression of miR-30e-5p in HK-2 cells and thus decreased Beclin1 levels to inhibit autophagy, but induced apoptosis, which may be the mechanism for CI-AKI.

Keywords:

Acute kidney injury, contrast, miR-30e-5p, autophagy, apoptosis, HK-2 cells.

Affiliation:

Department of Nephrology, Hongqi Hospital, Mudanjiang Medical University, 5 Tongxiang Road, Aimin District, Mudanjiang 157011, Heilongjiang, School of Public Health, North China University of Science and Technology, 57 Jianshe Road, Tangshan 063000, Hebei, Department of Nephrology, Hongqi Hospital, Mudanjiang Medical University, 5 Tongxiang Road, Aimin District, Mudanjiang 157011, Heilongjiang, Department of Nephrology, Hongqi Hospital, Mudanjiang Medical University, 5 Tongxiang Road, Aimin District, Mudanjiang 157011, Heilongjiang, Department of Nephrology, Hongqi Hospital, Mudanjiang Medical University, 5 Tongxiang Road, Aimin District, Mudanjiang 157011, Heilongjiang, Department of Nephrology, Hongqi Hospital, Mudanjiang Medical University, 5 Tongxiang Road, Aimin District, Mudanjiang 157011, Heilongjiang, Department of Ultrasonic Medicine, Hongqi Hospital, Mudanjiang Medical University, 5 Tongxiang Road, Aimin District, Mudanjiang 157011, Heilongjiang, Department of Thoracic Surgery, Tianjin Union Medical Center, Nankai University, 190 Jieyuan Road, Hongqiao District, Tianjin 300121, Tianjin



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