糖尿病性周围神经病理性疼痛中circ_SCN9A的作用与临床价值

THE ROLE OF CIRC_SCN9A IN THE PATHOGENESIS OF DIABETIC PERIPHERAL NEUROPATHIC PAIN AND ITS CLINICAL VALUE

摘要:

目的：研究钠电压门控通道α亚单位9（sodium voltage-gated channel alpha subunit 9，SCN9A）基因来源的环状RNA（circular RNA, circRNA）与糖尿病性周围神经病理性疼痛（diabetic peripheral neuropathic pain, DPNP）的关系和可能的分子机制。 方法：首先检测SCN9A基因来源的circRNA在DPNP患者血清中的表达谱；对差异circRNA进行细胞学定位研究；利用荧光素酶（Luciferase，LUC）报告基因实验检测上述circRNA对微小RNA（microRNA, miRNA）的吸附能力，ROC曲线（receiver operator characteristic curve,）评估circRNA在DPNP中的诊断价值；生物信息学分析并构建circRNA在DPNP中的作用通路并进行分子机制验证。 结果：SCN9A基因来源的hsa_circ_0117953（circ_SCN9A）在DPNP血清中显著高表达，在细胞内其主要定位于细胞质中，可调控多个miRNA的表达；circ_SCN9A在DPNP中具有良好的诊断价值且与疼痛程度相关；分子机制研究揭示circ_SCN9A调控的miRNA主要通过5-羟色胺能突触（Serotonergic synapse）途径发挥作用，这其中circ_SCN9A可通过吸附miR-1256而调控钾向内整流通道亚家族J6（potassium inwardly rectifying channel subfamily J member 6，KCNJ6）基因的蛋白表达。 结论：SCN9A基因来源的circ_SCN9A可通过Serotonergic synapse 途径参与DPNP的发生并可作为该病潜在的临床诊断生物学指标和治疗靶点。

Abstract:

Objective: To investigate the relationship and possible molecular mechanism of circular RNA (circRNA) derived from the gene of sodium voltage-gated channel α subunit 9 (SCN9A) with diabetic peripheral neuropathic pain (DPNP). Methods: Firstly, the expression profile of SCN9A-derived circRNAs in serum of DPNP patients was detected. Then, the cytological localization of screened differential circRNAs was studied. The microRNA (miRNA) spong capacity of the circRNAs was detected by luciferase (LUC) reporter assay, and the diagnostic value of circRNAs in DPNP was evaluated by receiver operator characteristic curve (ROC curve) model. Bioinformatics analysis was used to construct the functional pathway of circRNAs in DPNP and the molecular biological mechanism was also verified. Results: SCN9A derived hsa_circ_0117953 (circ_SCN9A) is significantly highly expressed in DPNP serum, and mainly located in the cytoplasm within the cell, regulating the expression of multiple miRNAs (miR-661, miR-1324, miR-877-3p, miR-1256, miR-153-3p, miR-1264) through the miRNA sponge mechanism. Circ_SCN9A had good diagnostic value in DPNP and is related to the levels of pain. Molecular mechanism study revealed multiple miRNAs that circ_SCN9A regulated mainly play a role by Serotonergic synapse pathway. Among them, circ_SCN9A can regulate the potassium inwardly rectifying channel subfamily J member 6 (KCNJ6) gene expression by sponging the miR-1256. Conclusion: SCN9A derived circ_SCN9A involved in DPNP development by Serotonergic synapse pathways and can be used as potential biological indicator in clinical diagnosis of DPNP.