miR-22 protect PC12 from ischemia/reperfusion-induced injury by targeting p53 upregulated modulator of apoptosis (PUMA).

Abstract

MicroRNAs have been implicated as critical regulatory molecules in many cerebrovascular diseases. Recent studies demonstrated miR-22 might provide a potential neuroprotective effect. However, the neuroprotective effect of miR-22 in ischemia/reperfusion (I/R) injury has not been thoroughly elucidated. In this study, the PC12 cells were subjected to 4 h oxygen and glucose deprivation (I) and 24 h reoxygenation (R). The PC12 cells were pre-transfected with miR-22 or anti-miR-22 or siRNA-mediated downregulation of p53-upregulated-modulator-of-apoptosis (PUMA)(PUMA siRNA) or their controls at 24 h prior to exposure to I/R. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot were employed to analyze mRNA and protein expression. PI and Annexin V assays and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay were used to quantify the rate of apoptosis. We found that miR-22 expression was significantly downregulated in the PC12 cells subjected to I/R. Loss of function of miR-22 increased PC12 apoptosis after I/R, and overexpression of miR-22 decreases PC12 apoptosis after I/R. PUMA protein was upregulated in the I/R group as compared with the sham group. The increased PUMA protein expression and apoptosis induced by I/R was reversed by transfection with PUMA siRNA. We concluded that I/R enhanced apoptosis and PUMA expression in PC12 cells via downregulation of miR-22. Enhanced miR-22 expression reversed both PUMA expression and apoptosis induced by I/R in PC12 cells. miR-22/PUMA axis has important implications for their clinical applications.

• Authors: Hongmei Jiao, Renyi Chen, Ziru Jiang, Lin Zhang, Hongwei Wang
• Journal: Bioengineered (Bioengineered) Vol. 11 Issue 1 Pg. 209-218 (12 2020) ISSN: 2165-5987 [Electronic] United States
• PMID: 32065044 (Publication Type: Journal Article)

Chemical References

• Apoptosis Regulatory Proteins
• Bbc3 protein, rat
• MIRN22 microRNA, rat
• MicroRNAs
• Tumor Suppressor Protein p53

Topics

• Animals
• Apoptosis
• Apoptosis Regulatory Proteins (genetics, metabolism)
• Blotting, Western
• In Situ Nick-End Labeling
• MicroRNAs (genetics, metabolism)
• Myocardial Reperfusion Injury (genetics, metabolism, pathology)
• PC12 Cells
• Rats
• Reperfusion Injury (genetics, metabolism, pathology)
• Tumor Suppressor Protein p53 (genetics, metabolism)