本文選題：依達(dá)拉奉 + 微小RNA-��； 參考：《中國(guó)病理生理雜志》2017年01期

【摘要】：目的:探討依達(dá)拉奉通過(guò)微小RNA-25(microRNA-25,miR-25)對(duì)高糖誘導(dǎo)的人神經(jīng)母細(xì)胞瘤SHSY5Y細(xì)胞凋亡的抑制作用及其機(jī)制。方法:將SH-SY5Y細(xì)胞用含高濃度葡萄糖的DMEM培養(yǎng)基和依達(dá)拉奉的聯(lián)合培養(yǎng)液共同培養(yǎng)24 h。MTT比色法測(cè)定SH-SY5Y細(xì)胞存活率;DCFH-DA熒光探針?lè)z測(cè)SH-SY5Y細(xì)胞中活性氧簇(ROS)的水平;采用流式細(xì)胞術(shù)檢測(cè)SH-SY5Y細(xì)胞的凋亡率;Western blot法檢測(cè)凋亡相關(guān)蛋白Bax和Bcl-2的表達(dá)水平;實(shí)時(shí)定量PCR檢測(cè)細(xì)胞中miR-25的表達(dá)水平。為進(jìn)一步闡明依達(dá)拉奉抑制高糖誘導(dǎo)的神經(jīng)細(xì)胞凋亡的作用靶點(diǎn),我們將miR-25抑制劑應(yīng)用于細(xì)胞,之后采用caspase-3凋亡試劑盒檢測(cè)細(xì)胞的凋亡率。結(jié)果:與對(duì)照組相比,高糖誘導(dǎo)后細(xì)胞存活率明顯降低,細(xì)胞中的ROS水平和細(xì)胞凋亡率明顯升高,Bax的表達(dá)明顯增加,Bcl-2的表達(dá)明顯降低,miR-25的表達(dá)水平也明顯降低。給予依達(dá)拉奉治療之后,細(xì)胞存活率明顯升高,ROS含量和細(xì)胞凋亡率明顯降低,Bax的蛋白水平明顯降低,Bcl-2蛋白水平明顯升高,miR-25的表達(dá)水平亦明顯升高。進(jìn)一步給予miR-25抑制劑后,caspase-3的水平明顯升高,此時(shí)同時(shí)給予依達(dá)拉奉后并不能抑制高糖引起的神經(jīng)細(xì)胞的凋亡。結(jié)論:依達(dá)拉奉對(duì)高糖誘導(dǎo)的SH-SY5Y細(xì)胞凋亡具有抑制作用,其作用靶點(diǎn)可能是miR-25。
[Abstract]:Aim: to investigate the inhibitory effect of Edaravone on the apoptosis of human neuroblastoma SHSY5Y cells induced by high glucose and its mechanism. Methods: SH-SY5Y cells were cultured in DMEM medium with high concentration of glucose and Edaravone co-cultured in 24 h.MTT colorimetric assay. The survival rate of SH-SY5Y cells was determined by fluorescence probe method. The level of Ros in SH-SY5Y cells was detected by fluorescence probe method. The apoptotic rate of SH-SY5Y cells was detected by flow cytometry and the expression of Bax and Bcl-2 was detected by Western blot, and the expression of miR-25 was detected by real-time quantitative PCR. To further elucidate the role of Edaravone in inhibiting the apoptosis of neurons induced by high glucose, we applied the miR-25 inhibitor to the cells, and then used the caspase-3 apoptosis kit to detect the apoptosis rate of the cells. Results: compared with the control group, the survival rate of the cells induced by high glucose was significantly decreased, the ROS level and the apoptosis rate of the cells were significantly increased, the expression of Bcl 2 was significantly increased and the expression level of miR-25 was significantly decreased. After treatment with Edaravone, the cell survival rate and apoptosis rate were significantly increased, the protein level of Bax was significantly decreased, the level of Bcl 2 protein was significantly increased, the expression level of miR-25 was significantly increased. The level of caspase-3 increased significantly after further administration of miR-25 inhibitor, but Edaravone did not inhibit the neuronal apoptosis induced by high glucose. Conclusion: Edaravone can inhibit the apoptosis of SH-SY5Y cells induced by high glucose, and its target may be miR-25.
【作者單位】： 中國(guó)人民解放軍第四零一醫(yī)院急診科;濱州醫(yī)學(xué)院附屬醫(yī)院老年內(nèi)科;
【分類(lèi)號(hào)】：R587.1
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本文編號(hào)：1985291