[No authors listed]
The present study aimed to investigate the role of microRNA (miR)â99bâ5p in spinal cord injury (SCI). Reverse transcriptionâquantitative polymerase chain reaction demonstrated that, compared with control mice, the expression levels of miRâ99bâ5p were upregulated in the mouse spinal cord following SCI. Mechanistic target of rapamycin (mTOR) was predicted to be the possible target of miRâ99bâ5p according to TargetScan and microrna databases. Dualâluciferase reporter assay verified that miRâ99bâ5p was able to target mTOR. Furthermore, the results of an apoptosis analysis demonstrated that there were few apoptotic neurons in the control group, whereas SCI induced a significant increase in the number of apoptotic cells. Conversely, apoptosis was inhibited following transfection with a miRâ99bâ5p inhibitor. The effects of miRâ99bâ5p on neurite growth were also evaluated. The results of an immunofluorescence analysis indicated that neurite growth was normal in the control group, whereas SCI induced a reduction in neurite growth, which was rescued following transfection with a miRâ99bâ5p inhibitor. The protein expression levels of mTOR were detected in the three groups by western blotting. The results demonstrated that, compared with the control group, the protein expression levels of mTOR were significantly reduced in SCI neurons, whereas transfection with a miRâ99bâ5p inhibitor suppressed the SCIâinduced reduction of mTOR. In conclusion, treatment with a miRâ99bâ5p inhibitor may attenuate SCIâinduced harmful alterations in spinal cord neurons via the regulation of mTOR expression.
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