依达拉奉右莰醇调控Nrf2/HO-1对实验性自身免疫性脑脊髓炎小鼠的保护作用

Effect of edaravone dexborneol on experimental autoimmune encephalomyelitis in mice by regulating Nrf2/HO-1

目的 依达拉奉通过抗氧化效应对实验性自身免疫性脑脊髓炎(EAE)小鼠具有保护作用,研究表明依达拉奉右莰醇抗氧化作用强于依达拉奉,文章主要探讨依达拉奉右莰醇对EAE小鼠的作用及分子机制。方法 将40只雌性C57BL/6小鼠采用随机数字表法分为空白对照组:注射等渗盐水;EAE模型组:制备EAE模型;依达拉奉右莰醇干预组:制备EAE模型+依达拉奉右莰醇干预(12.5 mg/kg);依达拉奉干预组:制备EAE模型+依达拉奉干预(10 mg/kg),每组10只。均采用腹腔注射给药,1次/d,连续14 d。观察小鼠行为学改变,并行神经功能障碍评分;HE和LFB染色观察脊髓组织病理改变;生化检测脑组织匀浆中氧化应激指标丙二醛(MDA)含量、活性氧(ROS)含量、超氧化物歧化酶(SOD)活性及过氧化氢酶(CAT)活性;Western blot检测脊髓组织中核因子E2相关因子2(Nrf2)和血红素加氧酶(HO-1)蛋白表达水平。结果 空白对照组均未发病,其余各组不同程度发病。与EAE模型组相比,依达拉奉右莰醇干预组及依达拉奉干预组发病潜伏期延长、进展期缩短、高峰期神经功能障碍评分降低,依达拉奉右莰醇干预组更明显(P<0.05)。EAE模型组脊髓组织大量炎性细胞浸润及大片髓鞘脱失,依达拉奉右莰醇干预组及依达拉奉干预组脊髓组织炎性细胞浸润及脱髓鞘程度减轻,依达拉奉右莰醇干预组最轻。与空白对照组MDA含量[(1.21±0.38)nmol/mg·pro],ROS含量[(361.9±58.24)ηnmol/mg·pro],SOD活性[(25.99±3.82)U/mg·pro]及CAT活性[(5.47±0.47)U/mg·pro]相比,EAE模型组MDA含量[(6.38±0.49)nmol/mg·pro]及ROS含量[(695.05±46.67)ηnmol/mg·pro]升高,SOD活性[(7.47±2.4)U/mg·pro]及CAT活性[(0.74±0.3)U/mg·pro]降低(P<0.05)。与EAE模型组相比,依达拉奉右莰醇干预组和依达拉奉干预组MDA含量[(2.26±0.35)、(3.44±0.29)nmol/mg·pro]及ROS含量[(468.96±27.05)、(584.69±66.12)ηnmol/mg·pro]降低,SOD活性[(18.45±1.41)、(13.14±2.76)U/mg·pro]及CAT活性[(3.96±0.18)、(1.76±0.28)U/mg·pro]升高,依达拉奉右莰醇干预组效果更明显(P<0.05)。与空白对照组相比,EAE模型组Nrf2和HO-1表达水平升高(P<0.05)。与EAE模型组相比,依达拉奉右莰醇干预组及依达拉奉干预组Nrf2和HO-1表达水平升高,依达拉奉右莰醇干预组更明显(P<0.05)。结论 依达拉奉右莰醇对EAE小鼠具有保护作用,其作用效果强于依达拉奉,其机制可能与依达拉奉右莰醇对Nrf2/HO-1具有更强的调控作用有关。

Objective Edaravone has a protective effect on experimental autoimmune encephalomyelitis(EAE) in mice through its antioxidant effect. Studies have shown that the antioxidant effect of edaravone dexborneol is stronger than that of edaravone. This article mainly discusses the effect and molecular mechanism of edaravone dexborneol on EAE mice. Methods Forty female C57 BL/6 mice were randomly divided into the blank control group: injection of isotonic saline;EAE model group: preparation of EAE model;edaravone dexborneol intervention group: preparation of EAE model+edaravone dexborneol intervention(12.5 mg/kg);edaravone intervention group: preparation of EAE model + edaravone intervention(10 mg/kg), with 10 rats in each group. All mice were given intraperitoneal injectiononce a dayfor 14 days. Behavioral changes and neurological dysfunction scores were observed. HE and LFB staining were used to observe the pathological changes of spinal cord tissue. The contents of malondialdehyde(MDA), reactive oxygen species(ROS), superoxide dismutase(SOD) and catalase(CAT) in brain tissue homogenate were determined by biochemical methods. The expression levels of nuclear factor E2-related factor 2(Nrf2) and HO1 protein in spinal cord tissue were detected by Western blot. Results There was no incidence in blank group and control group, and different degrees of incidence in other groups. Compared with the EAE model group, the incubation period was prolonged, the stage of progression was shortened, and the scores of peak neurological dysfunction were decreased in the edaravone dexborneol intervention group and edaravone intervention group, but more significantly in the former group(P<0.05).In the EAE model group, there were a large number of inflammatory cell infiltration and large myelin demyelination in the spinal cord tissue, but the degree in the edaravone dexborneol intervention group and edaravone intervention group was reduced, and the degree in the former group was the least.Compared with the MDA content [(1.21±0.38) nmol/mg.pro], ROS content [(361.9±58.24) ηnmol/mg.pro], SOD activity [(25.99±3.82) U/mg.pro] and CAT activity [(5.47±0.47) U/mg.pro] in the blank control group, the MDA content [(6.38±0.49) nmol/mg.pro] and ROS content [(695.05±46.67) ηnmol/mg.pro] increased, and SOD activity [(7.47±2.4) U/mg.pro] and CAT activity [(0.74±0.3) U/mg.pro] decreased(P<0.05) in EAE model group.Compared with the EAE model group, the MDA content [(2.26±0.35),(3.44±0.29) nmol/mg.pro] and ROS content [(468.96±27.05),(584.69±66.12) ηnmol/mg.pro] of edaravone dexborneol intervention group and edaravone intervention group were decreased. The SOD activity [(18.45±1.41),(13.14±2.76) U/mg.pro] and CAT activity [(3.96±0.18),(1.76±0.28) U/mg.pro] increased, and the effect of edaravone dexborneol intervention group was more obvious(P<0.05).Compared with blank control group, the expression levels of Nrf2 and HO-1 in EAE model group increased(P<0.05). Compared with EAE model group, the expression levels of Nrf2 and HO-1 in edaravone dexborneol intervention group and edaravone intervention group were increased, especially in the former group(P<0.05). Conclusion The protective effect of edaravone dexborneol on EAE mice was stronger than that of edaravone, and the mechanism may be related to the stronger regulation of Nrf2/HO-1 by edaravone.