miR-101 down-regulates mTOR expression and attenuates neuropathic pain in chronic constriction injury rat models.

We aimed to demonstrate the effects of microRNA (miR)-101 on neuropathic pain and explore the underlying mechanisms. Rat spinal microglia cells were isolated and inflammatory condition was stimulated by 24-h incubation with lipopolysaccharide (LPS). Rats were divided into 4 groups: sham, chronic constriction injury (CCI), CCI + miR-negative control (miR-NC) and CCI + miR-101 mimics. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) tests were conducted. The mRNA levels of key genes were determined by quantitative real-time polymerase chain reaction. Mammalian target of rapamycin (mTOR) protein level was detected by Western blot. Concentrations of interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α were examined by ELISA. MiR-101 was downregulated and mTOR was upregulated in lumbar spinal dorsal horns from CCI rats. Targetscan and luciferase reporter assay confirmed that mTOR was direct target of miR101. MiR-101 mimics inhibited LPS-stimulated increase in the levels of IL-6, IL-1β and TNF-α in primary microglial cells in vitro. In the rat CCI model, miR-101 mimics also suppressed CCI-induced decrease in PWT and PWL and inhibited CCI-induced increase in mRNA and protein levels of IL-6, IL-1β and TNF-α. In addition, miR-101 downregulated mTOR mRNA and protein expressions in CCI rats. Besides, CCI operation resulted in miR-101 downregulation and mTOR mRNA upregulation in spinal microglia cells in a time-dependent manner. In conclusion, miR-101 had neuropathic pain-attenuating activity through targeting mTOR.