富马酸二甲酯对β淀粉样蛋白诱导的氧化应激损伤的保护作用
发布时间：2018-11-17 15:20
【摘要】：目的:1.在Aβ诱导的星形胶质细胞和人神经母细胞瘤SH-SY5Y细胞的氧化应激损伤模型中,初步探讨Nrf2对Aβ诱导的氧化应激的抑制作用及DMF对Nrf2及其下游抗氧化应激因子如Nqo1和HO-1的调节作用。2.通过对星形胶质细胞Keap1、HDAC2因子和人神经母细胞瘤SH-SY5Y细胞Keap1、CX3CL1因子的检测,初步探讨DMF在Aβ诱导的氧化应激条件下对Nrf2作用机制。方法:1.将大鼠星形胶质细胞和人神经母细胞瘤SH-SY5Y细胞分别与Aβ25-35共培养,诱导氧化应激反应。在大鼠星形胶质细胞和人神经母细胞瘤SH-SY5Y细胞中分别设立Aβ组、Aβ+DMF组、Aβ+Si-nrf2组和Aβ+Si-nrf2+DMF组。Aβ组细胞给予Nrf2-conRNA转染。Aβ+Si-nrf2组细胞给予Nrf2-siRNA转染以敲低Nrf2基因的表达。以DMF(4uM)分别对Aβ组、Aβ+Si-nrf2两组细胞进行干预,干预后的细胞分别为Aβ+DMF组、Aβ+Si-nrf2+DMF组。然后采用RT-PCR分别检测两种细胞中各Aβ组、Aβ+DMF组、Aβ+Si-nrf2组和Aβ+Si-nrf2+DMF组的Nrf2、Nqo1、HO-1的mRNA表达水平。2.采用RT-PCR检测星形胶质细胞和人神经母细胞瘤SH-SY5Y细胞各Aβ组、Aβ+DMF组、Aβ+Si-nrf2组和Aβ+Si-nrf2+DMF组的Keap1 mRNA表达水平。采用Western blot检测星形胶质细胞各组的HDAC2蛋白表达水平和人神经母细胞瘤SH-SY5Y细胞各组的CX3CL1蛋白表达水平。结果:1.在星形胶质细胞和人神经母细胞瘤SH-SY5Y细胞中,Aβ+Si-nrf2组较Aβ组、Aβ+Si-nrf2+DMF组较Aβ+DMF组Nrf2、Nqo1和HO-1 mRNA表达水平显著降低(P0.05)。Aβ+DMF组较Aβ组Nrf2、Nqo1和HO-1 mRNA表达显著升高(P0.05)。2.在星形胶质细胞和人神经母细胞瘤SH-SY5Y中,Aβ+DMF组较Aβ组、Aβ+Si-nrf2+DMF组较Aβ+Si-nrf2组Keap1 m RNA表达显著降低(P0.05)。在星形胶质细胞中,Aβ+DMF组较Aβ组、Aβ+Si-nrf2+DMF组较Aβ+Si-nrf2组的HDAC2蛋白表达水平均明显下降(P0.05)。在人神经母细胞瘤SH-SY5Y细胞中,Aβ+DMF组较Aβ组、Aβ+Si-nrf2+DMF组较Aβ+Si-nrf2组的CX3CL1蛋白表达水平均明显升高(P0.05)。结论:1.Nrf2对Aβ诱导的氧化应激具有抑制作用。在Aβ诱导氧化应激的条件下,DMF可提高Nrf2的表达,增加抗氧化应激因子的表达。2.DMF可能通过抑制HDAC2的表达,提高Nrf2水平,从而减弱Aβ诱导的大鼠星形胶质细胞的氧化应激反应。DMF可能通过增加CX3CL1和减少Keap1的表达双重途径共同提高Nrf2水平,从而减弱Aβ诱导的人神经母细胞瘤SH-SY5H细胞的氧化应激反应。
[Abstract]:Objective: 1. In A 尾 -induced oxidative stress injury model of astrocytes and human neuroblastoma SH-SY5Y cells, To investigate the inhibitory effect of Nrf2 on A 尾 -induced oxidative stress and the regulatory effect of DMF on Nrf2 and its downstream antioxidant stress factors such as Nqo1 and HO-1. 2. Through the detection of astrocyte Keap1,HDAC2 factor and Keap1,CX3CL1 factor of human neuroblastoma SH-SY5Y cells, the mechanism of DMF acting on Nrf2 under A 尾 -induced oxidative stress was preliminarily investigated. Methods: 1. Rat astrocytes and human neuroblastoma SH-SY5Y cells were co-cultured with A 尾 25-35 to induce oxidative stress. In SH-SY5Y cells of rat astrocytes and human neuroblastoma, A 尾 group and A 尾 DMF group were established respectively. In A 尾 Si-nrf2 group and A 尾 Si-nrf2 DMF group, A 尾 cells were transfected with Nrf2-conRNA, and A 尾 Si-nrf2 cells were transfected with Nrf2-siRNA to lower the expression of Nrf2 gene. The cells of A 尾 group and A 尾 Si-nrf2 group were treated with DMF (4uM) respectively. The cells after intervention were A 尾 DMF group and A 尾 Si-nrf2 DMF group respectively. Then RT-PCR was used to detect the mRNA expression of Nrf2,Nqo1,HO-1 in A 尾 group, A 尾 DMF group, A 尾 Si-nrf2 group and A 尾 Si-nrf2 DMF group. The expression of Keap1 mRNA in astrocytes and human neuroblastoma SH-SY5Y cells was detected by RT-PCR in A 尾 group, A 尾 DMF group, A 尾 Si-nrf2 group and A 尾 Si-nrf2 DMF group. Western blot was used to detect the expression of HDAC2 protein in astrocytes and CX3CL1 protein in SH-SY5Y cells of human neuroblastoma. The result is 1: 1. In astrocytes and human neuroblastoma SH-SY5Y cells, A 尾 Si-nrf2 group was significantly lower than A 尾 group, A 尾 Si-nrf2 DMF group was significantly lower than A 尾 DMF group Nrf2,Nqo1 and HO-1 mRNA expression level (P0.05). A 尾 DMF group than A 尾 group Nrf2,) The expression of Nqo1 and HO-1 mRNA increased significantly (P0.05). In SH-SY5Y of astrocytes and human neuroblastoma, the expression of Keap1 m RNA in A 尾 DMF group was significantly lower than that in A 尾 group and A 尾 Si-nrf2 DMF group (P0.05). In astrocytes, the expression of HDAC2 protein in A 尾 DMF group was significantly lower than that in A 尾 group and A 尾 Si-nrf2 DMF group compared with A 尾 Si-nrf2 group (P0.05). In SH-SY5Y cells of human neuroblastoma, the expression of CX3CL1 protein in A 尾 DMF group was significantly higher than that in A 尾 group and A 尾 Si-nrf2 DMF group compared with A 尾 Si-nrf2 group (P0.05). Conclusion: 1.Nrf2 can inhibit the oxidative stress induced by A 尾. Under the condition of A 尾 -induced oxidative stress, DMF could increase the expression of Nrf2 and the expression of antioxidant stress factor. 2.DMF might increase the level of Nrf2 by inhibiting the expression of HDAC2. Thus attenuating the oxidative stress response of astrocytes induced by A 尾. DMF may increase the level of Nrf2 by increasing CX3CL1 and decreasing the expression of Keap1. Thus, the oxidative stress response of human neuroblastoma SH-SY5H cells induced by A 尾 was attenuated.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16
本文编号：2338290
[Abstract]:Objective: 1. In A 尾 -induced oxidative stress injury model of astrocytes and human neuroblastoma SH-SY5Y cells, To investigate the inhibitory effect of Nrf2 on A 尾 -induced oxidative stress and the regulatory effect of DMF on Nrf2 and its downstream antioxidant stress factors such as Nqo1 and HO-1. 2. Through the detection of astrocyte Keap1,HDAC2 factor and Keap1,CX3CL1 factor of human neuroblastoma SH-SY5Y cells, the mechanism of DMF acting on Nrf2 under A 尾 -induced oxidative stress was preliminarily investigated. Methods: 1. Rat astrocytes and human neuroblastoma SH-SY5Y cells were co-cultured with A 尾 25-35 to induce oxidative stress. In SH-SY5Y cells of rat astrocytes and human neuroblastoma, A 尾 group and A 尾 DMF group were established respectively. In A 尾 Si-nrf2 group and A 尾 Si-nrf2 DMF group, A 尾 cells were transfected with Nrf2-conRNA, and A 尾 Si-nrf2 cells were transfected with Nrf2-siRNA to lower the expression of Nrf2 gene. The cells of A 尾 group and A 尾 Si-nrf2 group were treated with DMF (4uM) respectively. The cells after intervention were A 尾 DMF group and A 尾 Si-nrf2 DMF group respectively. Then RT-PCR was used to detect the mRNA expression of Nrf2,Nqo1,HO-1 in A 尾 group, A 尾 DMF group, A 尾 Si-nrf2 group and A 尾 Si-nrf2 DMF group. The expression of Keap1 mRNA in astrocytes and human neuroblastoma SH-SY5Y cells was detected by RT-PCR in A 尾 group, A 尾 DMF group, A 尾 Si-nrf2 group and A 尾 Si-nrf2 DMF group. Western blot was used to detect the expression of HDAC2 protein in astrocytes and CX3CL1 protein in SH-SY5Y cells of human neuroblastoma. The result is 1: 1. In astrocytes and human neuroblastoma SH-SY5Y cells, A 尾 Si-nrf2 group was significantly lower than A 尾 group, A 尾 Si-nrf2 DMF group was significantly lower than A 尾 DMF group Nrf2,Nqo1 and HO-1 mRNA expression level (P0.05). A 尾 DMF group than A 尾 group Nrf2,) The expression of Nqo1 and HO-1 mRNA increased significantly (P0.05). In SH-SY5Y of astrocytes and human neuroblastoma, the expression of Keap1 m RNA in A 尾 DMF group was significantly lower than that in A 尾 group and A 尾 Si-nrf2 DMF group (P0.05). In astrocytes, the expression of HDAC2 protein in A 尾 DMF group was significantly lower than that in A 尾 group and A 尾 Si-nrf2 DMF group compared with A 尾 Si-nrf2 group (P0.05). In SH-SY5Y cells of human neuroblastoma, the expression of CX3CL1 protein in A 尾 DMF group was significantly higher than that in A 尾 group and A 尾 Si-nrf2 DMF group compared with A 尾 Si-nrf2 group (P0.05). Conclusion: 1.Nrf2 can inhibit the oxidative stress induced by A 尾. Under the condition of A 尾 -induced oxidative stress, DMF could increase the expression of Nrf2 and the expression of antioxidant stress factor. 2.DMF might increase the level of Nrf2 by inhibiting the expression of HDAC2. Thus attenuating the oxidative stress response of astrocytes induced by A 尾. DMF may increase the level of Nrf2 by increasing CX3CL1 and decreasing the expression of Keap1. Thus, the oxidative stress response of human neuroblastoma SH-SY5H cells induced by A 尾 was attenuated.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16
【参考文献】
相关期刊论文 前1条
1 刘理静;钱红;尹辉明;贺兼斌;张平;王在岩;;中华猕猴桃果仁非饱和脂肪酸通过激活Keap1/Nrf2信号通路增强肺纤维化大鼠抗氧化能力[J];细胞与分子免疫学杂志;2016年04期
，本文编号：2338290
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