The expression of Selenoprotein W(SelW)in C2C12 skeletal muscle cells was specifically decreased to examine its influence on the amount of glutathione(GSH)and the activity of glutathione peroxidase(GPx).SelW knock-dow...The expression of Selenoprotein W(SelW)in C2C12 skeletal muscle cells was specifically decreased to examine its influence on the amount of glutathione(GSH)and the activity of glutathione peroxidase(GPx).SelW knock-down was performed by RNA interference(RNAi)in cultured muscle cells and verified by Real-time PCR and Western blotting.In addition,cell viability,GSH content and GPx activity were assayed.The results showed that the mRNA level and protein expression of SelW were decreased successfully by 71.9%and 68.8%relative to control values,cell viability decreased by 21.5%,GSH increased by 29.76%,and GPx increased by 47.58%.WST assay showed that compared with blank control,the value of positive group dropped 21.5%;In GSH and GPx assay,compared with blank control the positive group increased29.76%and 47.58%separately.In conclusion,SelW knock-down by RNAi caused significant cytotoxity in skeletal muscle cells and led to compensatory increases in GSH content and GPx activity.These findings are consistent with the suggestions from bioinformatics indicating an antioxidative role for SelW in skeletal muscle cells.展开更多
为研究不同浓度亚硒酸钠对猪骨骼肌卫星细胞增殖分化以及硒蛋白W(Selenoprotein W,SelW)和硒结合蛋白1(Selenium-binding protein 1,SBP1)表达的影响。分别在0(对照组)、0.02(0.02%组)、0.1(0.1%组)、0.5(0.5%组)、2.5μg/mL(2.5%组)亚...为研究不同浓度亚硒酸钠对猪骨骼肌卫星细胞增殖分化以及硒蛋白W(Selenoprotein W,SelW)和硒结合蛋白1(Selenium-binding protein 1,SBP1)表达的影响。分别在0(对照组)、0.02(0.02%组)、0.1(0.1%组)、0.5(0.5%组)、2.5μg/mL(2.5%组)亚硒酸钠浓度的培养基条件下培养骨骼肌卫星细胞,在第1、2、3、4天观察细胞增殖分化情况,分别在第1、3、5、7天收集处理后的细胞。提取上述收集细胞的mRNA,检测SelW和SBP1基因表达。结果表明:1)从第1天到第4天,0.02%组、0.1%组、0.5%组细胞增殖与对照组相似,其中以0.1%组与对照组最接近。然而,2.5%组在第3天后降低了细胞数量,细胞出现大量死亡。2)不同浓度亚硒酸钠对SelW的表达在第3天影响最大,随着亚硒酸钠浓度增加呈现先增加后降低的趋势。第3天数据表明,与对照组相比,0.02%组、0.1%组、0.5%组和2.5%组均显著增加SelW的mRNA表达量,且0.1%组和0.5%的表达量显著高于0.02%组和2.5%组(P<0.05)。不同浓度亚硒酸钠对SBP1的表达在第3天和第7天影响最大,随着亚硒酸钠浓度增加呈现先增加后降低的趋势。在第3天时,与对照组相比,0.02%组和0.5%组显著增加SBP1的mRNA表达,且0.02%组组表达量显著高于0.1%组和2.5%组,0.5%组表达量显著高于0.1%组(P<0.05)。在第7天时,0.1%组和0.5组SBP1的mRNA表达量显著高于对照组、0.02%组和2.5%组,且2.5%组表达量显著低于对照组(P<0.05)。综上所述,0.02%和0.1%的低浓度亚硒酸钠不抑制和损害细胞增殖,而0.5%的亚硒酸钠水平会抑制细胞增殖速率,2.5%的亚硒酸钠水平有较强细胞毒性。0.02%和0.1%低浓度的亚硒酸钠可促进硒蛋白SelW和SBP1的mRNA表达,2.5%浓度亚硒酸钠抑制SelW和SBP1的mRNA表达。展开更多
目的:研究硒蛋白基因W和N与鸡肌肉品质的相关性。方法:选择1日龄科宝肉公鸡160只,随机分成低硒组和正常组,饲养42d,分别在14,28及42日龄取鸡的肌肉、肾脏、心脏组织和血清,测定GSH-Px和TAOC抗氧化指标,同时检测肌肉的p H值、肌纤维直径...目的:研究硒蛋白基因W和N与鸡肌肉品质的相关性。方法:选择1日龄科宝肉公鸡160只,随机分成低硒组和正常组,饲养42d,分别在14,28及42日龄取鸡的肌肉、肾脏、心脏组织和血清,测定GSH-Px和TAOC抗氧化指标,同时检测肌肉的p H值、肌纤维直径、滴水损失、MDA及硒蛋白基因Selw和Seln m RNA表达水平。结果:与正常组相比,低硒组鸡肌肉、肾脏、心脏和血清组织中GSH-Px活力与T-AOC能力显著降低(P<0.05);同时缺硒导致肌肉滴水损失增加(P<0.05),肌肉的肌纤维直径减少(P<0.05),p H值降低,MDA水平升高(P<0.05);硒缺乏下调了肌肉硒蛋白基因Selw及Seln m RNA表达水平(P<0.05)。结论 :根据相关性分析,肌肉硒蛋白基因Selw及Seln m RNA表达水平与鸡肉p H值及肌纤维直径有很好的相关性(P<0.05),而与滴水损失相关性不显著。展开更多
基金supported by China Postdoctoral Science Foundation(LRB06-262)Funds of Northeast Forestry University
文摘The expression of Selenoprotein W(SelW)in C2C12 skeletal muscle cells was specifically decreased to examine its influence on the amount of glutathione(GSH)and the activity of glutathione peroxidase(GPx).SelW knock-down was performed by RNA interference(RNAi)in cultured muscle cells and verified by Real-time PCR and Western blotting.In addition,cell viability,GSH content and GPx activity were assayed.The results showed that the mRNA level and protein expression of SelW were decreased successfully by 71.9%and 68.8%relative to control values,cell viability decreased by 21.5%,GSH increased by 29.76%,and GPx increased by 47.58%.WST assay showed that compared with blank control,the value of positive group dropped 21.5%;In GSH and GPx assay,compared with blank control the positive group increased29.76%and 47.58%separately.In conclusion,SelW knock-down by RNAi caused significant cytotoxity in skeletal muscle cells and led to compensatory increases in GSH content and GPx activity.These findings are consistent with the suggestions from bioinformatics indicating an antioxidative role for SelW in skeletal muscle cells.
文摘为研究不同浓度亚硒酸钠对猪骨骼肌卫星细胞增殖分化以及硒蛋白W(Selenoprotein W,SelW)和硒结合蛋白1(Selenium-binding protein 1,SBP1)表达的影响。分别在0(对照组)、0.02(0.02%组)、0.1(0.1%组)、0.5(0.5%组)、2.5μg/mL(2.5%组)亚硒酸钠浓度的培养基条件下培养骨骼肌卫星细胞,在第1、2、3、4天观察细胞增殖分化情况,分别在第1、3、5、7天收集处理后的细胞。提取上述收集细胞的mRNA,检测SelW和SBP1基因表达。结果表明:1)从第1天到第4天,0.02%组、0.1%组、0.5%组细胞增殖与对照组相似,其中以0.1%组与对照组最接近。然而,2.5%组在第3天后降低了细胞数量,细胞出现大量死亡。2)不同浓度亚硒酸钠对SelW的表达在第3天影响最大,随着亚硒酸钠浓度增加呈现先增加后降低的趋势。第3天数据表明,与对照组相比,0.02%组、0.1%组、0.5%组和2.5%组均显著增加SelW的mRNA表达量,且0.1%组和0.5%的表达量显著高于0.02%组和2.5%组(P<0.05)。不同浓度亚硒酸钠对SBP1的表达在第3天和第7天影响最大,随着亚硒酸钠浓度增加呈现先增加后降低的趋势。在第3天时,与对照组相比,0.02%组和0.5%组显著增加SBP1的mRNA表达,且0.02%组组表达量显著高于0.1%组和2.5%组,0.5%组表达量显著高于0.1%组(P<0.05)。在第7天时,0.1%组和0.5组SBP1的mRNA表达量显著高于对照组、0.02%组和2.5%组,且2.5%组表达量显著低于对照组(P<0.05)。综上所述,0.02%和0.1%的低浓度亚硒酸钠不抑制和损害细胞增殖,而0.5%的亚硒酸钠水平会抑制细胞增殖速率,2.5%的亚硒酸钠水平有较强细胞毒性。0.02%和0.1%低浓度的亚硒酸钠可促进硒蛋白SelW和SBP1的mRNA表达,2.5%浓度亚硒酸钠抑制SelW和SBP1的mRNA表达。
文摘目的:研究硒蛋白基因W和N与鸡肌肉品质的相关性。方法:选择1日龄科宝肉公鸡160只,随机分成低硒组和正常组,饲养42d,分别在14,28及42日龄取鸡的肌肉、肾脏、心脏组织和血清,测定GSH-Px和TAOC抗氧化指标,同时检测肌肉的p H值、肌纤维直径、滴水损失、MDA及硒蛋白基因Selw和Seln m RNA表达水平。结果:与正常组相比,低硒组鸡肌肉、肾脏、心脏和血清组织中GSH-Px活力与T-AOC能力显著降低(P<0.05);同时缺硒导致肌肉滴水损失增加(P<0.05),肌肉的肌纤维直径减少(P<0.05),p H值降低,MDA水平升高(P<0.05);硒缺乏下调了肌肉硒蛋白基因Selw及Seln m RNA表达水平(P<0.05)。结论 :根据相关性分析,肌肉硒蛋白基因Selw及Seln m RNA表达水平与鸡肉p H值及肌纤维直径有很好的相关性(P<0.05),而与滴水损失相关性不显著。
