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糖尿病机械性痛模型大鼠P2X3受体的时空表达
Spatio-temporal expression of P2X3 receptor in rats with diabetic mechanical allodynia
目的 探讨糖尿病机械性痛（DMA）模型大鼠中P2X3受体（P2X3R）的时空表达变化。方法 腹腔注射链脲菌素（STZ）建立大鼠DMA模型。采用von Frey细丝法测定DMA大鼠机械性痛阈，在不同时间点取腰髓4～5（L4～5）节段的脊髓背角（SDH）和背根神经节（DRG）以及足底皮肤，通过免疫荧光观察P2X3R阳性产物的表达变化。进一步Western blotting检测SDH和DRG中P2X3R蛋白的变化。结果 与对照组（CON组）相比，注射STZ 7d后大鼠机械性痛阈显著降低，在14d时达到最低并持续至28d（P<0.05）。免疫荧光和Western blotting结果均显示L4～5节段DRG中P2X3R表达在14d和21d时显著增多，与CON组相比差异有统计学意义（P<0.05），28d时恢复至CON组水平；而在SDH及皮肤中，P2X3R表达上调出现在21d和28d（P<0.05）。 结论 在STZ诱导的DMA大鼠机械性痛的不同时程中，DRG、SDH和皮肤中P2X3R的表达均呈现与痛敏变化几乎平行的变化趋势，但后两者比前者的变化在时间上略晚。这些结果说明P2X3R可能在DMA机械性痛敏的维持阶段起重要作用。
Objectiv To investigate the spatio-temporal expression of P2X3 receptor (P2X3R) in rats with diabetic mechanical allodynia (DMA).Methods DMA model in rats was induced by intraperitoneal injection of streptozocin (STZ). The von Frey filaments were applied to identify the changes of the paw withdrawal threshold (PWT) in DMA rats. Immunofluorescence was employed to detect the spatiotemporal expression of P2X3R in the spinal dorsal horn (SDH), dorsal root ganglion (DRG) and hind paw skin on different time points after intraperitoneal injection of STZ, respectively. The protein expression of P2X3R in SDH and DRG was further semi-quantitatively analyzed by Western blotting. Results Compared with control group, intraperitoneal injection STZ induced significant mechanical allodynia indicated by the reduced PWT from 7 days, and which reached the peak on 14d and maintained to 28days (P<0.05). The expression of P2X3R in DRG neurons was significantly increased on 14 days and 21 days (P<0.05), while that in SDH and skin was markedly increased on 21 days and 28 days, compared with control group (P<0.05). Conclusion With the progress of DMA, the expression of P2X3R was significantly increased in the SDH, DRG and skin, which was almost parallel with the mechanical allodynia, but the changes in SDH and skin were 1 week later than that in DRG. These results suggest that P2X3R may play a key role in the maintenance of the DMA.
糖尿病 / 机械性痛 / P2X3受体 / 脊髓背角 / 背根神经节 / 免疫荧光 / 免疫印迹法 / 大鼠
Diabetes mellitus /
Mechanical allodynia /
P2X3 receptor /
Spinal dorsal horn /
Dorsal root ganglion /
Immunofluorescence /
Western blotting /
Rat
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