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MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression post-transcriptionally by binding to their cognate target mRNAs. Emerging evidence suggests that miRNAs are critical regulators of neuronal functions. The expression pattern of miRNAs in the peripheral nervous system after peripheral nerve injury suggest that miRNAs may have important and yet unknown roles in the mechanisms of pain. Thus, we examined the role of miR-96 in neuropathic pain using a rat model of the condition chronic constriction sciatic nerve injury (CCI). We found that miR-96 alleviated neuropathic pain. The level of miR-96 was decreased within the ipsilateral dorsal root ganglion (DRG) after peripheral nerve injury but the Nav1.3 level was increased. Specifically, Intrathecal administration of miR-96 suppressed the expression of Nav1.3 induced by CCI. Further examination revealed that miR-96 inhibited the Nav1.3 mRNA expression in the embryonic DRG neurons in vitro. Our findings suggest that miR-96 participate in the regulation of neuropathic pain through inhibiting the expression of Nav1.3 in the DRG of CCI rats.