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目的:研究HMGB1(high mobility group box-1)在神经病理性痛大鼠脊髓水平的表达变化,探索HMGB1在神经病理性痛发生发展中的作用,为治疗神经病理性痛提供新的理论依据和治疗靶点。方法:(1)雄性SD大鼠(180～220)g 12只,随机均分为三组:NS组:鞘内注射生理盐水;A组:鞘内注射HMGB1 1μg;B组:鞘内注射HMGB1 10μg。盲法用von Frey测定给药前及给药后1 h、1、3、7、14、21、28 d大鼠50%机械缩足阈值(me-chanical withdrawal threshold,MWT);(2)雄性SD大鼠(180～220)g 5只,免疫荧光双标观察HMGB1在脊髓背角的表达定位;(3)雄性SD大鼠(180～220)g 42只,随机均分为对照组(6只),SNL模型组(每时间点6只),West-ern Blot方法观察大鼠脊髓背角HMGB1对照及术后1、3、7、14、21、28 d的表达变化。结果:(1)大鼠脊髓鞘内注射HMGB1后诱发长时程机械性痛敏,A组在鞘内给药后7 d MWT明显下降(P<0.01),B组给药后1 h MWT即显著下降,且持续存在至少28 d;(2)免疫荧光双标显示:HMGB1主要表达于NeuN标记的神经元,而GFAP阳性的星形胶质细胞以及OX42阳性的小胶质细胞几乎不表达HMGB1;(3)Western Blot结果显示,脊髓背角HMGB1在SNL模型术后缓慢增高,7 d时增高最为显著,且持续至少28 d。结论:以上结果表明,外周神经损伤后脊髓水平HMGB1的表达上调可能在神经病理性痛的产生和维持中起着重要作用。
OBJECTIVE: To investigate the expression of HMGB1 in spinal cord of rats with neuropathic pain and to explore the role of HMGB1 in the development of neuropathic pain and to provide a new theoretical basis and therapeutic target for the treatment of neuropathic pain . Methods: (1) 12 male SD rats (180 ~ 220) g were randomly divided into three groups: NS group: Intrathecal injection of saline; Group A: intrathecal HMGB1 1μg; Group B: intrathecal injection of HMGB1 10 μg. Blindness The von Frey’s 50% mechanical withdrawal threshold (MWT) before and 1, 3, 7, 14, 21, 28 d after administration were measured by blinded method. (2) Male SD rats (180 ~ 220) g 5, immunofluorescence double-labeled observation of HMGB1 expression in spinal dorsal horn positioning; (3) Male SD rats (180 ~ 220) g 42 were randomly divided into the control group Only), SNL model group (6 at each time point), West-ern Blot method to observe the expression of HMGB1 in spinal dorsal horn and 1,3,7,14,21,28 d after operation. Results: (1) The long-term mechanical hyperalgesia was induced by intrathecal injection of HMGB1 into the spinal cord of rats. The MWT of group A decreased significantly 7 days after intrathecal administration (P <0.01) (2) Double immunofluorescence staining showed that HMGB1 mainly expressed in NeuN-labeled neurons, whereas GFAP-positive astrocytes and OX42-positive microglia showed little or no expression of HMGB1 ; (3) Western Blot results showed that HMGB1 in dorsal horn of spinal cord increased slowly after SNL model, and increased most significantly at 7 d, and continued for at least 28 d. Conclusion: The above results suggest that upregulation of spinal cord HMGB1 after peripheral nerve injury may play an important role in the pathogenesis and maintenance of neuropathic pain.