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microRNA-212-3p attenuates neuropathic pain via targeting sodium voltage-gated channel alpha subunit 3 (NaV 1.3).

PURPOSE
To explorer the role and potential mechanism of miR-212-3p in neuropathic pain regulation.


METHODS
Adult male rats were used to establish chronic constriction injury (CCI) model to mimic the neuropathic pain. Then, paw withdrawal threshold (PWT), paw withdrawal thermal latency (PWL) was determined. Concentrations of interleukin 1 beta (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-α) was measured with enzyme-linked immune sorbent assay (ELISA) kit and expression of miR-212-3p was measured by real time quantitative PCR (RT-qPCR). Besides, miR-212-3p agomir was intrathecal injected into CCI rats and expression of key apoptotic proteins were determined by western blot. Furthermore, dual luciferase reporter assay was used to determine the binding of miR-212-3p and 3' untranslated region (3'UTR) of NaV1.3 and the expression levels of NaV1.3 were measured by western blot and RT-qPCR.


RESULTS
In CCI group, the PWT and PWL were significantly decreased and IL-1β, IL-6 and TNF-α were increased. miR-212-3p were decreased in response to CCI. Intrathecal injection of miR-212-3p agomir into CCI rats improved the PWT and PWL, decreased the IL-1β, IL-6 and TNF-α, decreased the expression levels of BCL2 associated X, apoptosis regulator (Bax), cleaved caspase-3 and increased the expression levels BCL2 apoptosis regulator (Bcl-2). The results of dual luciferase reporter assay showed miR-212-3p could directly bind with 3'UTR of NaV1.3. And the expression of NaV1.3 was up regulated in CCI rats intrathecal injected with miR-ctrl, whereas that was decreased in CCI rats intrathecal injected with miR-212-3p agomir.


CONCLUSION
Expression of miR-212a-3p attenuates neuropathic pain via targeting NaV1.3.