β-七叶皂苷通过抑制NF-κB、NLRP3信号通路改善大鼠创伤性脊髓损伤

β-seven leaf saponin improves traumatic spinal cord injury in rats by inhibiting NF-κB,NLRP3 signaling pathway

目的研究β-七叶皂苷对大鼠创伤性脊髓损伤(tSCI)的作用及其机制。方法将72只SD大鼠随机均分为假手术组、tSCI组、低剂量皂苷组和高剂量皂苷组。BV-2小胶质细胞分为对照组、模型组、低、中、高浓度皂苷组。采用旷场实验评估各组大鼠运动功能;HE染色观察各组大鼠脊髓组织病理学变化;ELISA法检测脊髓组织及BV-2细胞中炎症因子白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)表达。免疫荧光法检测BV-2细胞中核因子-κB(NF-κB)蛋白表达;Western blotting检测BV-2细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、IL-1β和Caspase-1蛋白表达。结果β-七叶皂苷能减轻tSCI大鼠脊髓组织病理损害程度,促进大鼠运动功能的恢复。与tSCI组或模型组比较,不同剂量或浓度皂苷组大鼠脊髓组织或BV-2细胞中IL-1β、IL-6和TNF-α表达水平均降低,且呈剂量或浓度依赖性降低(P<0.05)。与模型组比较,不同浓度皂苷组BV-2细胞中NF-κB、NLRP3、ASC、IL-1β和Caspase-1蛋白表达均降低,且呈浓度依赖性降低(P<0.05)。结论β-七叶皂苷对大鼠创伤性脊髓损伤具有一定的保护作用,该作用可能与抑制NF-κB/NLRP3信号通路有关。

Aim To study the effect and mechanism ofβ-seven leaf saponin on traumatic spinal cord injury(tSCI)in rats.Methods 72 SD rats were randomly divided into sham surgery group,tSCI group,and low-dose saponin group,and high-dose saponin group with 18 rats in each group.BV-2 microglial cells were divided into control group,model group,and low/medium/high concentration saponin groups.The motor function of rats in each group were evaluated by open field test.HE staining was used to observe the pathological changes of spinal cord tissue of rats in each group.The expression of inflammatory factors interleukin(IL)-1β,IL-6,tumor necrosis factor-α(TNF-α)in spinal cord tissue and BV-2 cells was detected by ELISA.The expression of nuclear factor-kappa B(NF-κB)protein in BV-2 cells was detected by immunofluorescence.Western blotting was used to detect the expression of NOD-like receptor thermal protein domain associated protein 3(NLRP3),apoptosis-associated speck-likeprotein containing(ASC),IL-1βand Caspase-1 in BV-2 cells.Resultsβ-seven leaf saponin could reduce the pathological damage of spinal cord tissue and promote the recovery of motor function in tSCI rats.Compared with the tSCI group or model group,the expression levels of IL-1β,IL-6 and TNF-αin spinal cord tissue of rats or BV-2 cells were dose-dependently decreased after treatment with different doses or concentrations of saponins(P<0.05).Compared with the model group,the expression of NF-κB,NLRP3,ASC,IL-1βand Caspase-1 proteins in BV-2 cells was concentrations-dependently decreased after treatment with different concentrations of saponin(P<0.05).Conclusionβ-seven leaf saponin has a protective effect on traumatic spinal cord injury in rats,which may be related to the inhibition of NF-κB/NLRP3 signaling pathway.