Role of miR-145 in chronic constriction injury in rats
Xiaolin Pang 1 , Yuanzhang Tang 2 , Dongya Zhang 1
- 1 Department of Anesthesiology, First Hospital of Tsinghua University, Beijing 100016, P.R. China.
- 2 Department of Pain Management, Xuanwu Hospital of Capital Medical University, Beijing 100053, P.R. China.
The present study aims to investigate the effects and underlying mechanisms of miRNA-145 (miR-145) in rat models of chronic constriction injury (CCI). Rats were randomly divided into control, sham, CCI, agomiRNA (agomiR)-normal control (NC) and agomiR-145 groups (n=25 in each group); in addition, 30 rats with CCI were divided into small hairpin (sh)RNA-NC and shRNA-ras responsive element binding protein 1 (RREB1) groups. Paw withdrawal thermal latency (PWTL) and paw withdrawal mechanical threshold (PWMT) were detected. Reverse transcription-quantitative polymerase chain reaction was used to detect miR-145 expression levels, and western blotting was performed to measure RREB1 and phosphorylated-protein kinase B (p-AKT) expression levels. In addition, a dual luciferase reporter assay was conducted to identify the target gene of miR-145. PWMT and PWTL were decreased in CCI rats and this decrease was alleviated by miR-145 injection. At 1, 3, 5 and 7 days after CCI, miR-145 expression level in the spinal cord tissue of rats in the CCI group was significantly decreased compared with 1 day before CCI (P<0.05). Compared with the CCI group, miR-145 expression level in the agomiR-145 group was significantly higher (P<0.05). In addition, expression levels of RREB1 and p-AKT were significantly increased in the CCI group and significantly decreased in the agomiR-145 group (P<0.05). Furthermore, knockdown of RREB1 expression by shRNA-RREB1 significantly increased values of PWMT and PWTL, decreased expression levels of RREB1 and p-AKT, and increased miR-145 expression levels (P<0.05). Further investigation demonstrated that miR-145 can bind with RREB1 mRNA. In conclusion, miR-145 may be involved in the development of CCI through regulating the expression of RREB1.
Keywords: chronic constriction injury; microRNA-145; phosphorylated protein kinase B; ras responsive element binding protein 1.