目的筛选影响胆固醇调节元件结合蛋白1a(cholesterol regulatory element binding protein,SREBP1a)蛋白稳定性的去泛素化酶,并探索其调控机制。方法通过去泛素化酶库筛选显著影响SREBP1a表达的去泛素化酶,免疫蛋白印记实验和实时荧光定...目的筛选影响胆固醇调节元件结合蛋白1a(cholesterol regulatory element binding protein,SREBP1a)蛋白稳定性的去泛素化酶,并探索其调控机制。方法通过去泛素化酶库筛选显著影响SREBP1a表达的去泛素化酶,免疫蛋白印记实验和实时荧光定量PCR(qRT-PCR)评估去泛素化酶对SREBP1a以及升脂基因表达的影响;通过红色荧光蛋白标记人源低密度脂蛋白(human Dil-low density lipoprotein,Human Dil-LDL)摄取和油红O染色等实验技术检测细胞摄取低密度脂蛋白(LDL)和脂质沉积情况。结果去泛素化酶库筛选发现泛素特异肽酶37(ubiquitin specific peptidase 37,USP37)可显著增加肝细胞SREBP1a蛋白表达水平,促进胆固醇摄取及脂质沉积。USP37基因敲除可显著降低SREBP1a蛋白表达水平,抑制升脂基因表达及脂质沉积。结论去泛素化酶USP37通过稳定SREBP1a蛋白表达,促进胆固醇摄取及脂质沉积,揭示了SREBP1a翻译后调控的新模式。展开更多
目的探讨前列腺癌组织中固醇调节元件结合蛋白1(SREBP1)、乙酰辅酶A羧化酶α(ACCα)的表达及对前列腺癌细胞系DU145增殖、迁移和侵袭的影响。方法通过在线网站TIMER、UALCAN、GEPIA和THE HUMAN PROTEIN ATLAS分析SREBP1和ACCα在前列腺...目的探讨前列腺癌组织中固醇调节元件结合蛋白1(SREBP1)、乙酰辅酶A羧化酶α(ACCα)的表达及对前列腺癌细胞系DU145增殖、迁移和侵袭的影响。方法通过在线网站TIMER、UALCAN、GEPIA和THE HUMAN PROTEIN ATLAS分析SREBP1和ACCα在前列腺癌中表达情况。收集合肥医科大学第五附属(珠海)医院2016年1月至2018年12月期间收治的58例前列腺癌(PCa)和58例良性前列腺增生(BPH)患者石蜡标本,采用免疫组织化学法检测上述组织中SREBP1和ACCα的表达;采用shRNA下调DU145细胞中SREBP1的表达后,qRT-PCR检测ACCα的表达变化;采用CCK-8法检测细胞增殖能力;流式细胞仪检测细胞周期分布;Western blot实验检测SREBP1和ACCα的表达;通过Transwell迁移和侵袭实验检测沉默SREBP1后DU145细胞迁移和侵袭的变化;细胞划痕愈合实验检测干扰SREBP1后对DU145细胞划痕愈合能力的影响;EdU实验检测沉默SREBP1表达后细胞增殖能力;通过油红O染色,观察干扰SREBP1的表达后,前列腺癌细胞中脂质含量变化。结果TIMER、UALCAN在线数据库分析发现,同正常前列腺组织相比,ACCα在前列腺癌中表达显著升高(P<0.001)。沉默SREBP1后,DU145细胞增殖、迁移和侵袭能力均较对照组显著降低(P<0.01);流式细胞术检测沉默SREBP1后DU145细胞发生G1期阻滞(P<0.01)。SREBP1敲低后前列腺癌细胞中脂肪含量显著降低(P<0.01)。结论SREBP1可能通过调控ACCα的表达,促进前列腺癌细胞增殖、迁移和侵袭。展开更多
Objective:To investigate the effects of quercetin on inflammatory signaling pathways and hepatic oxidative injury using a streptozotocin-induced liver injury model.Methods:Four groups of 32 rats were used in this stud...Objective:To investigate the effects of quercetin on inflammatory signaling pathways and hepatic oxidative injury using a streptozotocin-induced liver injury model.Methods:Four groups of 32 rats were used in this study:three groups were given streptozotocin to induce diabetes,and one group was given a normal control.For treatment groups,each group received either metformin(200 mg/kg body weight)or quercetin(50 mg/kg body weight)for a month.The expression of SREBP1c was detected by quantitative RT-PCR and fibrosis-related proteins(TGF-βand p-Smad3)was evaluated by Western blot.Furthermore,MCP and IL-1βwere determined by ELISA.Results:Quercetin significantly reduced insulin and glucose levels.Besides,it reduced the serum levels of ALT,AST,and ALP,improved lipid profile,lowered the MDA level,increased SOD activity,decreased the rise in MCP-1 and IL-1β,inhibited the TGF-β/Smad3 signaling pathway,decreasedα-SMA and SREBP1c expression,and increased AMPK(P<0.05).Conclusions:Quercetin could significantly mitigate hepatic damage by modulating the expression of pro-inflammatory cytokines and fibrosis markers.展开更多
文摘目的筛选影响胆固醇调节元件结合蛋白1a(cholesterol regulatory element binding protein,SREBP1a)蛋白稳定性的去泛素化酶,并探索其调控机制。方法通过去泛素化酶库筛选显著影响SREBP1a表达的去泛素化酶,免疫蛋白印记实验和实时荧光定量PCR(qRT-PCR)评估去泛素化酶对SREBP1a以及升脂基因表达的影响;通过红色荧光蛋白标记人源低密度脂蛋白(human Dil-low density lipoprotein,Human Dil-LDL)摄取和油红O染色等实验技术检测细胞摄取低密度脂蛋白(LDL)和脂质沉积情况。结果去泛素化酶库筛选发现泛素特异肽酶37(ubiquitin specific peptidase 37,USP37)可显著增加肝细胞SREBP1a蛋白表达水平,促进胆固醇摄取及脂质沉积。USP37基因敲除可显著降低SREBP1a蛋白表达水平,抑制升脂基因表达及脂质沉积。结论去泛素化酶USP37通过稳定SREBP1a蛋白表达,促进胆固醇摄取及脂质沉积,揭示了SREBP1a翻译后调控的新模式。
文摘目的探讨前列腺癌组织中固醇调节元件结合蛋白1(SREBP1)、乙酰辅酶A羧化酶α(ACCα)的表达及对前列腺癌细胞系DU145增殖、迁移和侵袭的影响。方法通过在线网站TIMER、UALCAN、GEPIA和THE HUMAN PROTEIN ATLAS分析SREBP1和ACCα在前列腺癌中表达情况。收集合肥医科大学第五附属(珠海)医院2016年1月至2018年12月期间收治的58例前列腺癌(PCa)和58例良性前列腺增生(BPH)患者石蜡标本,采用免疫组织化学法检测上述组织中SREBP1和ACCα的表达;采用shRNA下调DU145细胞中SREBP1的表达后,qRT-PCR检测ACCα的表达变化;采用CCK-8法检测细胞增殖能力;流式细胞仪检测细胞周期分布;Western blot实验检测SREBP1和ACCα的表达;通过Transwell迁移和侵袭实验检测沉默SREBP1后DU145细胞迁移和侵袭的变化;细胞划痕愈合实验检测干扰SREBP1后对DU145细胞划痕愈合能力的影响;EdU实验检测沉默SREBP1表达后细胞增殖能力;通过油红O染色,观察干扰SREBP1的表达后,前列腺癌细胞中脂质含量变化。结果TIMER、UALCAN在线数据库分析发现,同正常前列腺组织相比,ACCα在前列腺癌中表达显著升高(P<0.001)。沉默SREBP1后,DU145细胞增殖、迁移和侵袭能力均较对照组显著降低(P<0.01);流式细胞术检测沉默SREBP1后DU145细胞发生G1期阻滞(P<0.01)。SREBP1敲低后前列腺癌细胞中脂肪含量显著降低(P<0.01)。结论SREBP1可能通过调控ACCα的表达,促进前列腺癌细胞增殖、迁移和侵袭。
文摘Objective:To investigate the effects of quercetin on inflammatory signaling pathways and hepatic oxidative injury using a streptozotocin-induced liver injury model.Methods:Four groups of 32 rats were used in this study:three groups were given streptozotocin to induce diabetes,and one group was given a normal control.For treatment groups,each group received either metformin(200 mg/kg body weight)or quercetin(50 mg/kg body weight)for a month.The expression of SREBP1c was detected by quantitative RT-PCR and fibrosis-related proteins(TGF-βand p-Smad3)was evaluated by Western blot.Furthermore,MCP and IL-1βwere determined by ELISA.Results:Quercetin significantly reduced insulin and glucose levels.Besides,it reduced the serum levels of ALT,AST,and ALP,improved lipid profile,lowered the MDA level,increased SOD activity,decreased the rise in MCP-1 and IL-1β,inhibited the TGF-β/Smad3 signaling pathway,decreasedα-SMA and SREBP1c expression,and increased AMPK(P<0.05).Conclusions:Quercetin could significantly mitigate hepatic damage by modulating the expression of pro-inflammatory cytokines and fibrosis markers.
