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目的探讨鞘内转染白细胞介素-10(IL-10)基因对神经病理性疼痛大鼠的镇痛作用。方法雌性SD大鼠,体重230-250 g,坐骨神经压榨性损伤(CCI)后3 d出现热痛觉过敏大鼠随机分为基因治疗组(pcDNA3．1-IL-10组)、真核表达载体组(pcDNA3．1组)、生理盐水组(NS组)和CCI组(n =12),前3组分别经蛛网膜下腔注射pcDNA3．1-IL-10/脂质体复合物、pcDNA3．1/脂质体复合物或生理盐水30μl,另取12只雌性SD大鼠为假手术组(Sham组),仅暴露右侧坐骨神经而不予压榨,亦不行蛛网膜下腔注射。转染后2、7、7、10、14 d热辐射法测定大鼠热痛阈;RT-PCR检测转染后3 d坐骨神经、腰段脊髓、海马组织IL-10 mRNA的表达,ELISA法检测上述组织、血浆及转染后1-10 d脑脊液中IL- 10水平;细胞毒性法检测转染后14 d坐骨神经、腰段脊髓、海马和血浆TNF-α水平。结果转染后2 -14 d CCI组热痛阈长于Sham组,而pcDNA3．1-IL-10能逆转热痛阈延长;pcDNA3．1-IL-10组转染后3d 坐骨神经、腰段脊髓、海马等组织可检测到IL-10 mRNA,且IL-10含量升高,脑脊液IL-10浓度在转染后1-7 d升高,转染后14 d坐骨神经、腰段脊髓、海马等TNF-α含量降低(P<0．05)。结论鞘内转染IL-10能通过抑制炎症反应在一定程度上减轻大鼠神经病理性疼痛。
Objective To investigate the analgesic effects of intrathecal transfection of interleukin-10 (IL-10) gene on neuropathic pain rats. Methods Female SD rats weighing 230-250 g and thermal hyperalgesia 3 days after sciatic nerve crush injury (CCI) were randomly divided into gene therapy group (pcDNA3.1-IL-10 group), eukaryotic expression vector group (pcDNA3.1 group), saline group (NS group) and CCI group (n = 12). The first three groups were injected subcutaneously with pcDNA3.1-IL-10 / liposome complex and pcDNA3.1 / Liposome complex or normal saline 30μl, another 12 female SD rats were sham operation group (Sham group), only the right sciatic nerve was exposed without being squeezed, nor subarachnoid injection. The heat pain threshold was measured at 2, 7, 7, 10, 14 days after transfection. The mRNA expression of IL-10 in sciatic nerve, lumbar spinal cord and hippocampus was detected by RT-PCR 3 days after transfection. The expression of IL- The levels of IL-10 in cerebrospinal fluid of the above tissues, plasma and 1-10 days after transfection were measured. The levels of TNF-α in the sciatic nerve, lumbar spinal cord, hippocampus and plasma on day 14 after transfection were detected by cytotoxicity assay. Results The thermal pain threshold of CCI group was longer than that of Sham group 2 to 14 days after transfection, while the thermal threshold of thrombosis was reversed by pcDNA3.1-IL-10. In the pcDNA3.1-IL-10 group, the sciatic nerve, lumbar spinal cord, IL-10 mRNA and IL-10 levels were detected in the hippocampus and hippocampus, IL-10 concentration in cerebrospinal fluid increased 1-7 days after transfection, and the level of IL- α content decreased (P <0.05). Conclusion Intrathecal transfection of IL-10 can relieve the neuropathic pain to a certain extent in rats by inhibiting the inflammatory reaction.