Selenium is a crucial trace element that contributes to physiological processes in the body as selenoproteins.Selenoproteins serve as an integral role in the body in controlling the redox state of cells and protecting...Selenium is a crucial trace element that contributes to physiological processes in the body as selenoproteins.Selenoproteins serve as an integral role in the body in controlling the redox state of cells and protecting against damage induced by oxidative stress.This study aimed to investigate the effects and possible mechanism of selenium on selenoproteins expression in EA.hy926 cells induced by oxidized low density lipoprotein(oxLDL).The impact of selenium on the viability of EA.hy926 cells was detected by the methylthiazolyldiphenyltetrazolium bromide(MTT)method,and intracellular reactive oxygen species(ROS)level and mitochondrial membrane potential were assessed by fluorescent probe DCFH-DA and JC-1,respectively.RNA-seq,quantitative real-time polymerase chain reaction(qPCR),and Western blot were used to investigate the selenoprotein expression.Selenoprotein mRNA translation efficiency was analyzed by ribosome profiling(Ribo-Seq)coupled with transcriptomics.Our data showed that selenium supplementation(0.5μmol/L)significantly decreased ROS production,increased mitochondrial inner membrane potential and increased the proliferative activity of EA.hy926 cells induced by oxLDL.Moreover,The protective effects of selenium against oxLDL-induced EA.hy926 cell injury were associated with the upregulation of the expressions of selenoproteins glutathione peroxidase 1(GPX1),glutathione peroxidase 4(GPX4),and thioredoxin reductase 1(TXNRD1).Furthermore,the expressions of selenoproteins GPX1 and GPX4 were hierarchically controlled,but the expressions of selenoproteins TXNRD1 were mainly regulated by oxLDL.Finally,Ribo-Seq coupled with transcriptomics results demonstrated that the expressions of selenoproteins GPX1,GPX4,and TXNRD1 were regulated at the translation process level.These findings suggested that selenium could have preventive effects in oxLDL induced EA.hy926 cell injury by regulating the selenoprotein expression,and the selenoproteins expressions at the translation level in vascular endothelial cells need further study.展开更多
Flavonoid 3’-hydroxylase(F3’ H) and 3’,5’-hydroxylase(F3’5’ H) generate precursor molecules for the synthesis of cyanidin-based anthocyanins(red) and delphinidin-based anthocyanins(blue to purple) in Ribes nigru...Flavonoid 3’-hydroxylase(F3’ H) and 3’,5’-hydroxylase(F3’5’ H) generate precursor molecules for the synthesis of cyanidin-based anthocyanins(red) and delphinidin-based anthocyanins(blue to purple) in Ribes nigrum L.(black currant). In this study, full-length 1780 and 165’-bp cDNA homologs of RnF3’5’ H1 and RnF3’ H1 from black currant were identified and cloned using a homologous cloning technique. Data revealed that Rn F3’5’ H1 and RnF3’ H1 are homologs that encode enzymes involved in anthocyanin synthesis from different plants,which phylogenetically cluster with the CYP75 B and CYP75 A families in the P45’ superfamily, respectively.The enzymes encoded by these two genes also shared a high homology with flavonoid hydroxylases identified from other plants. Furthermore, RnF3’5’ H1 and RnF3’ H1 levels were upregulated during fruit maturation. RnF3’5’ H1 levels were associated with both anthocyanin and soluble carbohydrate levels in blackcurrant, while RnF3’ H1 expression did not have such an association. The structure and expression patterns of RnF3’5’ H1 and RnF3’ H1 in blackcurrant were also characterized. Further studies should aid understanding of anthocyanin biosynthesis in black currant to develop molecular approaches and manipulate anthocyanin production in blackcurrant.展开更多
基金supported by grants from the National Natural Science Foundation of China(NSFC,81960588)the Ningxia Natural Science Foundation(2020AAC03146)support from the Ningxia Medical University。
文摘Selenium is a crucial trace element that contributes to physiological processes in the body as selenoproteins.Selenoproteins serve as an integral role in the body in controlling the redox state of cells and protecting against damage induced by oxidative stress.This study aimed to investigate the effects and possible mechanism of selenium on selenoproteins expression in EA.hy926 cells induced by oxidized low density lipoprotein(oxLDL).The impact of selenium on the viability of EA.hy926 cells was detected by the methylthiazolyldiphenyltetrazolium bromide(MTT)method,and intracellular reactive oxygen species(ROS)level and mitochondrial membrane potential were assessed by fluorescent probe DCFH-DA and JC-1,respectively.RNA-seq,quantitative real-time polymerase chain reaction(qPCR),and Western blot were used to investigate the selenoprotein expression.Selenoprotein mRNA translation efficiency was analyzed by ribosome profiling(Ribo-Seq)coupled with transcriptomics.Our data showed that selenium supplementation(0.5μmol/L)significantly decreased ROS production,increased mitochondrial inner membrane potential and increased the proliferative activity of EA.hy926 cells induced by oxLDL.Moreover,The protective effects of selenium against oxLDL-induced EA.hy926 cell injury were associated with the upregulation of the expressions of selenoproteins glutathione peroxidase 1(GPX1),glutathione peroxidase 4(GPX4),and thioredoxin reductase 1(TXNRD1).Furthermore,the expressions of selenoproteins GPX1 and GPX4 were hierarchically controlled,but the expressions of selenoproteins TXNRD1 were mainly regulated by oxLDL.Finally,Ribo-Seq coupled with transcriptomics results demonstrated that the expressions of selenoproteins GPX1,GPX4,and TXNRD1 were regulated at the translation process level.These findings suggested that selenium could have preventive effects in oxLDL induced EA.hy926 cell injury by regulating the selenoprotein expression,and the selenoproteins expressions at the translation level in vascular endothelial cells need further study.
基金supported by Heilongjiang Natural Science Foundation(Grant No.C2016015)with a project titled‘‘Biological function analysis of F3050H gene of black currant(Ribes nigrum L.)’’
文摘Flavonoid 3’-hydroxylase(F3’ H) and 3’,5’-hydroxylase(F3’5’ H) generate precursor molecules for the synthesis of cyanidin-based anthocyanins(red) and delphinidin-based anthocyanins(blue to purple) in Ribes nigrum L.(black currant). In this study, full-length 1780 and 165’-bp cDNA homologs of RnF3’5’ H1 and RnF3’ H1 from black currant were identified and cloned using a homologous cloning technique. Data revealed that Rn F3’5’ H1 and RnF3’ H1 are homologs that encode enzymes involved in anthocyanin synthesis from different plants,which phylogenetically cluster with the CYP75 B and CYP75 A families in the P45’ superfamily, respectively.The enzymes encoded by these two genes also shared a high homology with flavonoid hydroxylases identified from other plants. Furthermore, RnF3’5’ H1 and RnF3’ H1 levels were upregulated during fruit maturation. RnF3’5’ H1 levels were associated with both anthocyanin and soluble carbohydrate levels in blackcurrant, while RnF3’ H1 expression did not have such an association. The structure and expression patterns of RnF3’5’ H1 and RnF3’ H1 in blackcurrant were also characterized. Further studies should aid understanding of anthocyanin biosynthesis in black currant to develop molecular approaches and manipulate anthocyanin production in blackcurrant.
