Objective:To explore the effects of histone deacetylase 6(HDAC-6) on the PD cell model induced by proteasome inhibitor lactacystin.Methods:Human neuroblastoma SK-N-SH cells were cultured.The wild type pcDNA3.1-alpha-s...Objective:To explore the effects of histone deacetylase 6(HDAC-6) on the PD cell model induced by proteasome inhibitor lactacystin.Methods:Human neuroblastoma SK-N-SH cells were cultured.The wild type pcDNA3.1-alpha-synuclein eukaryotic expression plasmid was transferred into the cells which then were divided into control group,group L,group T and group T+L.The eells of group L were added with 5 umol/L lactacystin dissolved in dimethylsulfoxide(DMSO) to induce PD cell model with abnormal protein aggregation,the cells of control group were treated with 5 umol/L DMSO,the cells of group T were treated with 5 umol/L selective HDAC-6 inhibitor tubacin dissolved in DMSO,and the cells of group T+L were treated with 5 umol/L lactaeystin and 10 umol/L tubacin dissolved in DMSO.The expression levels of alpha-synuclein oligomers,HSP-27 and HSP-70 were detected by Western blot and the cell survival rate of all the groups was detected by MTT colorimetric assay,and compared 24 h after the cells were treated.Results:The expression levels of alpha-synuclein oligomers,HSP-27 and HSP-70 of the cells of group L were significantly higher than the control group,and the cell survival rate was significantly lower(P<0.05);the expression level of alpha-synuclein oligomers of the cells of group T+L was significantly higher than group L,but the expression level of HSP-27 and HSP-70 were significantly lower,and so as the cell survival rate(P<0.05);the differences of the expression level of alpha-synuclein oligomers,HSP-27 and HSP-70 and the cell survival rate of the cells of group T and the control group were not statistically significant(P>0.05).Conclusions:The expression level of alphasynuclein oligomers can be improved and the cell survival rate can be reduced by the PD cell model induced by lactacystin and treated with selective HDAC-6 inhibitor tubacin,which means that alpha-synuclein oligomers of the PD cell model induced by lactacystin can be inhibited and the cell survival rate can be improved by HDAC-6,and the mechanism may be related to the increased of HSP-27 and HSP-70.展开更多
As Bortezomib and Carfilzomib are approved to treat refractory multiple myeloma, 20 S proteasome has become a highly interested tumor therapeutic target. Because of drug resistance of Bortezomib and Carfilzomib, β-la...As Bortezomib and Carfilzomib are approved to treat refractory multiple myeloma, 20 S proteasome has become a highly interested tumor therapeutic target. Because of drug resistance of Bortezomib and Carfilzomib, β-lactone natural products are used to treat cancer, arthritis, asthma, and Alzheimer’s diseases. Now the second generation drug candidates are developed, eg. Salinosporamide A（1）. This review is aiming to encapsulate the synthetic methods of β-lactone proteasome inhibitors, lactacystin（3） and its analogs.展开更多
文摘目的:探讨人参皂苷Rg1(ginsenoside Rg1,Rg1)对lactacystin所致SH-SY5Y细胞损伤的保护作用及可能机制。方法:实验分正常对照组、lactacystin组、lactacystin和Rg1联合处理组(0.1,1,10μmol/L),分别用CCK-8检测细胞活力,免疫荧光法、流式细胞术测定法检测二价金属离子转运蛋白1(divalent metal transporter 1,DMT1)表达情况,calcein-AM荧光检测细胞的摄铁能力,JC-1染色检测细胞线粒体膜电位,荧光探针carboxy-H2DCFDA检测胞内氧化应激水平。结果:与lactacystin组相比,lactacystin和Rg1联合处理组的细胞活力显著增加,细胞DMT1表达减少,细胞摄铁能力下降,线粒体膜电位增加,胞内氧化应激水平降低,差异均有统计学意义(P<0.05)。结论:Rgl对lactacystin诱导损伤的SHSY5Y细胞具有一定的神经保护作用,其作用机制可能与下调DMT1介导的细胞摄铁能力,稳定线粒体膜电位,降低氧化应激反应有关。
基金supported by Natural Science Foundation of Guangdong Province:(gd28182334)
文摘Objective:To explore the effects of histone deacetylase 6(HDAC-6) on the PD cell model induced by proteasome inhibitor lactacystin.Methods:Human neuroblastoma SK-N-SH cells were cultured.The wild type pcDNA3.1-alpha-synuclein eukaryotic expression plasmid was transferred into the cells which then were divided into control group,group L,group T and group T+L.The eells of group L were added with 5 umol/L lactacystin dissolved in dimethylsulfoxide(DMSO) to induce PD cell model with abnormal protein aggregation,the cells of control group were treated with 5 umol/L DMSO,the cells of group T were treated with 5 umol/L selective HDAC-6 inhibitor tubacin dissolved in DMSO,and the cells of group T+L were treated with 5 umol/L lactaeystin and 10 umol/L tubacin dissolved in DMSO.The expression levels of alpha-synuclein oligomers,HSP-27 and HSP-70 were detected by Western blot and the cell survival rate of all the groups was detected by MTT colorimetric assay,and compared 24 h after the cells were treated.Results:The expression levels of alpha-synuclein oligomers,HSP-27 and HSP-70 of the cells of group L were significantly higher than the control group,and the cell survival rate was significantly lower(P<0.05);the expression level of alpha-synuclein oligomers of the cells of group T+L was significantly higher than group L,but the expression level of HSP-27 and HSP-70 were significantly lower,and so as the cell survival rate(P<0.05);the differences of the expression level of alpha-synuclein oligomers,HSP-27 and HSP-70 and the cell survival rate of the cells of group T and the control group were not statistically significant(P>0.05).Conclusions:The expression level of alphasynuclein oligomers can be improved and the cell survival rate can be reduced by the PD cell model induced by lactacystin and treated with selective HDAC-6 inhibitor tubacin,which means that alpha-synuclein oligomers of the PD cell model induced by lactacystin can be inhibited and the cell survival rate can be improved by HDAC-6,and the mechanism may be related to the increased of HSP-27 and HSP-70.
基金National Natural Science Foundation of China(Grant No.21362021)Inner Mongolia Natural Science Foundation(Grant No.2015MS0206)
文摘As Bortezomib and Carfilzomib are approved to treat refractory multiple myeloma, 20 S proteasome has become a highly interested tumor therapeutic target. Because of drug resistance of Bortezomib and Carfilzomib, β-lactone natural products are used to treat cancer, arthritis, asthma, and Alzheimer’s diseases. Now the second generation drug candidates are developed, eg. Salinosporamide A（1）. This review is aiming to encapsulate the synthetic methods of β-lactone proteasome inhibitors, lactacystin（3） and its analogs.
