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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。
Objective To investigate the temporal and spatial expression of P2X3R in diabetic rats with mechanical pain (DMA). Methods The rat model of DMA was established by intraperitoneal injection of streptozotocin (STZ). The mechanical pain threshold of DMA rats was determined by von Frey filament method. Spinal cord dorsal horn (SDH) and dorsal root ganglion (DRG) at 4 ~ 5 (L4 ~ 5) The changes of the expression of P2X3R-positive products were observed by immunofluorescence. The changes of P2X3R protein in SDH and DRG were further detected by Western blotting. Results Compared with the control group (CON group), the mechanical pain threshold was significantly decreased after 7 days of STZ injection, reaching the lowest level at 14 days and continuing until 28 days (P <0.05). Immunofluorescence and Western blotting results showed that the expression of P2X3R in L4 ~ 5 segment of DRG increased significantly at 14d and 21d, which was significantly different from that in CON group (P <0.05), and recovered to CON group at 28d. In SDH and skin, P2X3R expression increased 21d and 28d (P <0.05). Conclusions The expression of P2X3R in DRG, SDH and skin is almost parallel to the change of pain sensitivity in different time course of mechanical pain induced by STZ in DMA rats. However, the change of P2X3R in DRG, late. These results suggest that P2X3R may play an important role in the maintenance phase of DMA mechanical hyperalgesia.