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目的观察鞘内注射甘珀酸(CBX)对腰5脊神经切断(SNT)大鼠机械痛阈及脊髓背角星形胶质细胞标志物(GFAP)和TNF-α、IL-1β表达的影响。方法 36只成年♂SD大鼠,随机分成3组(n=12),假手术组(Sham组)、SNT+NS组(NS组)、SNT+CBX(CBX组)。Sham组仅暴露腰5脊神经,不切断,NS组和CBX组切断腰5脊神经。术后10d,Sham组和NS组鞘内注射生理盐水10μl,CBX组鞘内注射甘珀酸5μg(10μl),分别于术前1 d,术后1、3、5、7、10 d及给药后1、2、4、6 h随机取6只大鼠测损伤侧机械痛阈(MWT),并采用免疫组化技术观察给药后2 h后损伤侧脊髓背角中GFAP表达的变化,ELISA测定损伤侧脊髓背角TNF-α和IL-1β的水平。结果 3组大鼠术前1 d,MWT差异均无显著性(P>0.05);Sham组术后各时间点与术前相比损伤侧MWT差异无显著性(P>0.05);与Sham组相比,NS组和CBX组术后1、3、5、7、10 d痛阈明显下降(P<0.05);NS组术后10 d给药后1、2、4、6 h,MWT与给药前无差异(P>0.05),但给药后2 h损伤侧脊髓背角GFAP的表达和TNF-α、IL-1β的水平较Sham组明显增高(P<0.05);CBX组给药后1、2、4 h损伤侧MWT较给药前明显提高(P<0.05),6 h无差异(P>0.05),给药后2 h损伤侧脊髓背角GFAP的表达和TNF-α、IL-1β的水平较NS组明显降低(P<0.05)。结论单次鞘内注射5μg CBX可改善脊神经切断大鼠损伤侧机械痛敏行为,该效应可能与其抑制损伤侧脊髓背角星形胶质细胞的活化,减少TNF-α、IL-1β的释放有关。
Objective To observe the effects of intrathecal carbenicillin (CBX) on mechanical pain threshold, expression of glial fibrillary acidic protein (GFAP), TNF-α and IL-1β in lumbar spinal dorsal root ganglion (SNT) rats. Methods Thirty-six adult male SD rats were randomly divided into 3 groups (n = 12), Sham group, SNT + NS group (NS group) and SNT + CBX group (CBX group). Sham group only exposed lumbar 5 spinal nerves, not cut off, NS group and CBX group cut lumbar 5 spinal nerves. At 10 days after operation, Sham group and NS group received intrathecal injection of 10μl normal saline and CBX group received intrathecal injection of 5μg (10μl) of carbenoxolol, respectively, at 1 day, 1, 3, 5, 7 and 10 days after operation Six rats were randomly selected at 1, 2, 4 and 6 h after drug administration to measure the mechanical damage threshold (MWT). The expression of GFAP in spinal dorsal horn was observed by immunohistochemistry 2 h after injury. The levels of TNF-α and IL-1β in injured spinal dorsal horn were measured by ELISA. Results There was no significant difference in MWT at 1 day before operation between the three groups (P> 0.05). There was no significant difference in the MWT at the injury side between Sham group and Sham group Compared with the NS group and the CBX group, the pain threshold was significantly decreased at 1, 3, 5, 7 and 10 days after operation (P <0.05) However, there was no difference before administration (P> 0.05), but the expression of GFAP and the levels of TNF-α and IL-1β in dorsal horn of spinal dorsal horn at 2 h after injury were significantly higher than those in Sham group At 1, 2 and 4 h after injury, the MWT at the injury side was significantly increased (P <0.05) and no difference at 6 h (P> 0.05). The expression of GFAP and TNF-α, IL-1β levels were significantly lower than NS group (P <0.05). Conclusion A single intrathecal injection of 5 μg CBX may improve the mechanical hyperalgesia in spinal cord injury rats, which may be related to the inhibition of astrocyte activation and the release of TNF-α and IL-1β in injured spinal dorsal horn .