富馬酸二甲酯對β淀粉樣蛋白誘導的氧化應激損傷的保護作用
發(fā)布時間：2018-11-17 15:20
【摘要】：目的:1.在Aβ誘導的星形膠質(zhì)細胞和人神經(jīng)母細胞瘤SH-SY5Y細胞的氧化應激損傷模型中,初步探討Nrf2對Aβ誘導的氧化應激的抑制作用及DMF對Nrf2及其下游抗氧化應激因子如Nqo1和HO-1的調(diào)節(jié)作用。2.通過對星形膠質(zhì)細胞Keap1、HDAC2因子和人神經(jīng)母細胞瘤SH-SY5Y細胞Keap1、CX3CL1因子的檢測,初步探討DMF在Aβ誘導的氧化應激條件下對Nrf2作用機制。方法:1.將大鼠星形膠質(zhì)細胞和人神經(jīng)母細胞瘤SH-SY5Y細胞分別與Aβ25-35共培養(yǎng),誘導氧化應激反應。在大鼠星形膠質(zhì)細胞和人神經(jīng)母細胞瘤SH-SY5Y細胞中分別設立Aβ組、Aβ+DMF組、Aβ+Si-nrf2組和Aβ+Si-nrf2+DMF組。Aβ組細胞給予Nrf2-conRNA轉(zhuǎn)染。Aβ+Si-nrf2組細胞給予Nrf2-siRNA轉(zhuǎn)染以敲低Nrf2基因的表達。以DMF(4uM)分別對Aβ組、Aβ+Si-nrf2兩組細胞進行干預,干預后的細胞分別為Aβ+DMF組、Aβ+Si-nrf2+DMF組。然后采用RT-PCR分別檢測兩種細胞中各Aβ組、Aβ+DMF組、Aβ+Si-nrf2組和Aβ+Si-nrf2+DMF組的Nrf2、Nqo1、HO-1的mRNA表達水平。2.采用RT-PCR檢測星形膠質(zhì)細胞和人神經(jīng)母細胞瘤SH-SY5Y細胞各Aβ組、Aβ+DMF組、Aβ+Si-nrf2組和Aβ+Si-nrf2+DMF組的Keap1 mRNA表達水平。采用Western blot檢測星形膠質(zhì)細胞各組的HDAC2蛋白表達水平和人神經(jīng)母細胞瘤SH-SY5Y細胞各組的CX3CL1蛋白表達水平。結(jié)果:1.在星形膠質(zhì)細胞和人神經(jīng)母細胞瘤SH-SY5Y細胞中,Aβ+Si-nrf2組較Aβ組、Aβ+Si-nrf2+DMF組較Aβ+DMF組Nrf2、Nqo1和HO-1 mRNA表達水平顯著降低(P0.05)。Aβ+DMF組較Aβ組Nrf2、Nqo1和HO-1 mRNA表達顯著升高(P0.05)。2.在星形膠質(zhì)細胞和人神經(jīng)母細胞瘤SH-SY5Y中,Aβ+DMF組較Aβ組、Aβ+Si-nrf2+DMF組較Aβ+Si-nrf2組Keap1 m RNA表達顯著降低(P0.05)。在星形膠質(zhì)細胞中,Aβ+DMF組較Aβ組、Aβ+Si-nrf2+DMF組較Aβ+Si-nrf2組的HDAC2蛋白表達水平均明顯下降(P0.05)。在人神經(jīng)母細胞瘤SH-SY5Y細胞中,Aβ+DMF組較Aβ組、Aβ+Si-nrf2+DMF組較Aβ+Si-nrf2組的CX3CL1蛋白表達水平均明顯升高(P0.05)。結(jié)論:1.Nrf2對Aβ誘導的氧化應激具有抑制作用。在Aβ誘導氧化應激的條件下,DMF可提高Nrf2的表達,增加抗氧化應激因子的表達。2.DMF可能通過抑制HDAC2的表達,提高Nrf2水平,從而減弱Aβ誘導的大鼠星形膠質(zhì)細胞的氧化應激反應。DMF可能通過增加CX3CL1和減少Keap1的表達雙重途徑共同提高Nrf2水平,從而減弱Aβ誘導的人神經(jīng)母細胞瘤SH-SY5H細胞的氧化應激反應。
[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.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.16
本文編號：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.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.16
【參考文獻】
相關期刊論文 前1條
1 劉理靜;錢紅;尹輝明;賀兼斌;張平;王在巖;;中華獼猴桃果仁非飽和脂肪酸通過激活Keap1/Nrf2信號通路增強肺纖維化大鼠抗氧化能力[J];細胞與分子免疫學雜志;2016年04期
，本文編號：2338290
本文鏈接：http://sikaile.net/yixuelunwen/jsb/2338290.html
最近更新
教材專著
