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目的:研究脊神经结扎后,外周CD4+T细胞迁移浸润至腰段脊髓及其分子调控机制。方法:选择健康成年清洁级雄性SD大鼠,随机分为脊神经结扎组(Tx组)、假冒手术组(S组)、对照组(C组),用up-down方法测定50%机械缩足阈值(50%MWT),并用FACS检测腰段脊髓CD4+T细胞的浸润情况,RT-qPCR法测定脊髓中趋化因子CCL2、CCL5、CXCL10mRNA的表达水平,ELISA检测血清中相关细胞因子的含量。结果:与S组、C组相比,术后3 d,Tx组50%MWT明显降低(P<0.01),至术后14 d达最低值,S组与C组在各时间点均无明显差异(P>0.05);术后7 d,Tx组腰段脊髓浸润的CD4+T细胞明显增加(P<0.01),同时伴有CCL2、CCL5mRNA的表达增加(P<0.05);术后14 d,Tx组浸润的CD4+T细胞较术后7 d减少,虽高于S组、C组,但无统计学意义,上述各趋化因子的表达水平也无明显差异;术后7 d、14 d,血清中各细胞因子的含量三组均无统计学意义。结论:神经损伤后,脊髓中高表达的趋化因子促进了外周CD4+T细胞的中枢浸润,脊髓浸润的CD4+T细胞可能与神经病理性疼痛的维持有关。
OBJECTIVE: To study the molecular mechanisms of migration and infiltration of peripheral CD4 + T cells into the lumbar spinal cord after spinal nerve ligation. Methods: Healthy adult male Sprague Dawley rats were randomly divided into spinal nerve ligation group (Tx group), sham operation group (S group) and control group (C group). The 50% mechanical contracting threshold (50% MWT). The infiltration of CD4 + T cells in lumbar spinal cord was detected by FACS. The expressions of chemokines CCL2, CCL5 and CXCL10 in the spinal cord were measured by RT-qPCR. The levels of cytokines in serum were detected by ELISA. Results: Compared with S group and C group, the 50% MWT of Tx group decreased significantly (P <0.01) 3 days after operation and reached the lowest value on the 14th day after operation. There was no significant difference between S group and C group at each time point (P <0.05). At 7 days after operation, the infiltration of CD4 + T cells in the lumbar spinal cord of Tx group was significantly increased (P <0.01), accompanied by increased expression of CCL2 and CCL5 mRNA (P < , While the infiltration of CD4 + T cells in Tx group was less than that of the 7th day postoperatively, although it was higher than those in S and C groups. However, there was no significant difference in the expression levels of these chemokines. d, serum cytokines in the three groups were not statistically significant. CONCLUSION: Chemotactic factors highly expressed in the spinal cord promote central infiltration of peripheral CD4 + T cells after nerve injury. Infiltration of spinal cord CD4 + T cells may be related to the maintenance of neuropathic pain.