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目的:探讨丙戊酸(VPA)对大鼠脊髓损伤(SCI)后运动功能恢复的影响及作用机制。方法:60只雄性SD大鼠随机均分为3组:假手术组(C组)、损伤组(SCI组)和丙戊酸保护组(VPA组)。SCI组和VPA组采用改良Allen法制作大鼠T10 SCI模型。VPA组术后即刻及其后每12h皮下注射VPA 300mg/kg,至取材;C组和SCI组在相应时间点注射等体积的生理盐水。伤后6h,每组取5只大鼠处死取材,其余大鼠在伤后24h、48h和72h每组取5只先行后肢运动功能BBB评分,随后处死取材。切片后分别行HE染色观察脊髓组织病理变化,免疫荧光双标法在激光共聚焦显微镜下观察核因子κB(NF-κB)途径的激活状态,免疫组化法检测白介素1β(IL-1β)的表达。结果:C组大鼠各时间点BBB评分均为21分,VPA组和SCI组各时间点的评分均低于C组(P<0.05),但VPA组各时间点的评分均高于同时间点SCI组,在伤后48h和72h两组差异有显著性(P<0.05)。病理检查显示C组脊髓组织形态正常,VPA组和SCI组伤后6h损伤中央区即可见明显出血灶,灰质中神经元肿胀坏死,白质中神经纤维肿胀;伤后24h、48h出血灶界限更明显,并可见空洞形成和炎症细胞浸润;伤后72h上述病理变化仍明显;VPA组各时间点的病理变化与SCI组相似,但炎症细胞浸润减少。C组偶见或未见NF-κB核阳性细胞和IL-1β表达,与C组相比,SCI组和VPA组NF-κB核阳性细胞百分比和IL-1β表达量从伤后6h即显著性增高,24h达高峰,以后逐渐减少,72h仍显著性高于C组(P<0.05);VPA组各时间点NF-κB核阳性细胞百分比和IL-1β表达量均低于同时间点SCI组(P<0.05)。结论:VPA可促进大鼠SCI后运动神经功能恢复,其机制可能与抑制炎症反应有关。
Objective: To investigate the effect and mechanism of valproic acid (VPA) on recovery of motor function after spinal cord injury (SCI) in rats. Methods: Sixty male SD rats were randomly divided into three groups: sham operation group (C group), injury group (SCI group) and valproate protective group (VPA group). The T10 SCI model of rats was made by modified Allen method in SCI group and VPA group. The VPA group was subcutaneously injected VPA 300mg / kg immediately after the operation and afterwards. The rats in Group C and SCI were given equal volume of saline at the corresponding time point. Five rats in each group were sacrificed at 6 hours after injury. Five rats in each group were sacrificed at 24h, 48h and 72h after injury respectively. The sections were stained with HE staining to observe the histopathological changes of spinal cord. Immunofluorescence double labeling method was used to observe the activation of nuclear factor kappa B (NF-κB) pathway by laser scanning confocal microscopy. The expression of interleukin-1β expression. Results: The score of BBB at each time point in group C was 21, and the score at each time point in VPA group and SCI group was lower than that in group C (P <0.05), but the score of VPA group at each time point was higher than that at the same time At SCI group, there was significant difference between the two groups at 48h and 72h after injury (P <0.05). The histopathological examination showed that the morphology of spinal cord in group C was normal. In the VPA group and the SCI group, obvious hemorrhagic lesions were seen in the central area 6 h after injury, swelling and necrosis of neurons in gray matter, and swelling of nerve fibers in white matter; , And the formation of voids and infiltration of inflammatory cells were observed. The pathological changes were still obvious at 72h after injury. The pathological changes at different time points in VPA group were similar to those in SCI group, but infiltration of inflammatory cells was decreased. Compared with group C, the percentage of NF-κB positive cells and the expression of IL-1β in SCI group and VPA group were significantly higher than those in group C (P <0.05). The percentage of NF-κB positive cells and the expression of IL-1β at each time point in VPA group were lower than those in SCI group at the same time point (P <0.05). Conclusion: VPA can promote motor nerve function recovery after SCI in rats, which may be related to the inhibition of inflammatory reaction.