miR-99b-5p靶向FGFR3对紫杉醇诱导的慢性神经痛的形成影响

mir-99b-5P targeting FGFR3 inhibits the chronic neuralgia

摘要:

目的 探讨miR-99b-5p对紫杉醇诱导的大鼠慢性神经痛的影响。方法 成年SD雄性大鼠分为空白组、紫杉醇模型组、agomiR-99b-5p治疗组和agomiR-NC对照组。采用隔天腹腔注射紫杉醇(2mg/kg,共四次)的方法制备紫杉醇诱导的慢性神经痛大鼠模型。通过vonFrey纤维丝和热辐射法检测大鼠机械缩足阈值(MWT)和热缩足潜伏期(TWL)；RT-qPCR检测miR-99b-5p的表达判断agomiR-99b-5p的激活作用和背根神经节中炎症因子TNF-α、IL-6和IL-1β含量的变化；Western blot检测背根神经节FGFR3的变化；双荧光素酶报告基因实验检测miR-99b-5p对FGFR3的靶向作用。结果 agomiR-99b-5p能显著增加大鼠背根神经节miR-99b-5p的含量,降低紫杉醇诱导的慢性神经痛大鼠的MWT和TWL,同时也能显著下调背根神经节的炎症因子水平。荧光素酶结果表明相对于NC mimic组,miR-99b-5p mimic能够抑制FGFR3的表达。结论 miR-99b -5p可能通过靶向抑制FGFR3进而抑制背根神经节炎症因子的含量达到降低紫杉醇诱导大鼠慢性神经疼痛的目的。

Abstract:

Aim To investigate the effect of miR-99b-5p on paclitaxel-induced chronic neuralgia in rats. Methods Male SD rats were divided into blank group, paclitaxel model group, agomiR-99b-5p group and agomiR-NC group. Paclitaxel-induced chronic neuralgia rats were prepared by intraperitoneal injection of paclitaxel (2mg/kg, four times in total) every other day. Detection of mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) in these rats by vonFrey filament and thermal radiation. The expression of miR-99b-5p, TNF-α, IL-6 and IL-1β in dorsal root ganglia of rats were detected by RT-qPCR assay. Western blot detection of FGFR3, and luciferase assay was used to detect the targeting effect of miR-99b-5p on FGFR3. Results agomiR-99b-5p can significantly increase the content of miR-99b-5p in rat dorsal root ganglia, reduce the MWT and TWL of paclitaxel-induced chronic neuralgia rats, and significantly down-regulate the levels of inflammatory factors in dorsal root ganglia. The luciferase experiment indicated that miR-99b-5p mimic could inhibit FGFR3 expression compared to the NC mimic group. Conclusion miR-99b-5p may reduce paclitaxel-induced chronic neuralgia in rats by targeting FGFR3 inhibition and further inhibiting the content of inflammatory factors in dorsal root ganglia