Liu Jian-Tao, Zhang Si, Gu Bing, Li Hua-Nan, Wang Shuo-Yu, Zhang Shui-Yin
Jiangxi Key Laboratory of Bioprocess Engineering, Jiangxi Science & Technology Normal University, Nanchang, Jiangxi Province, China.
Department of Spine Surgery, Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi Province, China.
Neural Regen Res. 2017 Sep;12(9):1507-1518. doi: 10.4103/1673-5374.215263.
Methylprednisolone is a commonly used drug for the treatment of spinal cord injury, but high doses of methylprednisolone can increase the incidence of infectious diseases. Methotrexate has anti-inflammatory activity and immunosuppressive effects, and can reduce inflammation after spinal cord injury. To analyze gene expression changes and the molecular mechanism of methotrexate combined with methylprednisolone in the treatment of spinal cord injury, a rat model of spinal cord contusion was prepared using the PinPoint™ precision cortical impactor technique. Rats were injected with methylprednisolone 30 mg/kg 30 minutes after injury, and then subcutaneously injected with 0.3 mg/kg methotrexate 1 day after injury, once a day, for 2 weeks. TreadScan gait analysis found that at 4 and 8 weeks after injury, methotrexate combined with methylprednisolone significantly improved hind limb swing time, stride time, minimum longitudinal deviation, instant speed, footprint area and regularity index. Solexa high-throughput sequencing was used to analyze differential gene expression. Compared with methylprednisolone alone, differential expression of 316 genes was detected in injured spinal cord treated with methotrexate and methylprednisolone. The 275 up-regulated genes were mainly related to nerve recovery, anti-oxidative, anti-inflammatory and anti-apoptotic functions, while 41 down-regulated genes were mainly related to proinflammatory and pro-apoptotic functions. These results indicate that methotrexate combined with methylprednisolone exhibited better effects on inhibiting the activity of inflammatory cytokines and enhancing antioxidant and anti-apoptotic effects and thereby produced stronger neuroprotective effects than methotrexate alone. The 316 differentially expressed genes play an important role in the above processes.
甲基泼尼松龙是治疗脊髓损伤常用的药物，但大剂量甲基泼尼松龙会增加传染病的发生率。甲氨蝶呤具有抗炎活性和免疫抑制作用，可减轻脊髓损伤后的炎症。为分析甲氨蝶呤联合甲基泼尼松龙治疗脊髓损伤的基因表达变化及分子机制，采用PinPoint™精确皮质撞击器技术制备大鼠脊髓挫伤模型。大鼠在损伤后30分钟注射30mg/kg甲基泼尼松龙，然后在损伤后1天皮下注射0.3mg/kg甲氨蝶呤，每天1次，共2周。TreadScan步态分析发现，在损伤后4周和8周，甲氨蝶呤联合甲基泼尼松龙显著改善了后肢摆动时间、步幅时间、最小纵向偏差、即时速度、足迹面积和规律性指数。采用Solexa高通量测序分析差异基因表达。与单独使用甲基泼尼松龙相比，在用甲氨蝶呤和甲基泼尼松龙治疗的损伤脊髓中检测到316个基因的差异表达。275个上调基因主要与神经恢复、抗氧化、抗炎和抗凋亡功能相关，而41个下调基因主要与促炎和促凋亡功能相关。这些结果表明，与单独使用甲氨蝶呤相比，甲氨蝶呤联合甲基泼尼松龙在抑制炎性细胞因子活性、增强抗氧化和抗凋亡作用方面表现出更好的效果，从而产生更强的神经保护作用。316个差异表达基因在上述过程中起重要作用。
