Selenoprotein S Attenuates Tumor Necrosis Factor-α-Induced Dysfunction in Endothelial Cells

Figure 3

Effect of SelS on the viability of HUVECs and NO and eNOS expressions. (a) Viability of the transfected HUVECs after a 12 h stimulation with TNF-α (100 ng/ml), tested using the CCK-8 method. (b) The concentrations of NO in transfected HUVECs with 6 h TNF-α (10 ng/ml) treatment, determined using the nitrate reductase assay. The protein expression of eNOS in 10 ng/ml TNF-α-induced HUVECs after transfection with SelS plasmid (c) or SelS siRNA (d) examined by the western blot. The mRNA levels of eNOS in 10 ng/ml TNF-α-induced HUVECs after transfection with SelS plasmid (e) or SelS siRNA (f). The cells were transfected with pcDNA3.1-SelS recombinant plasmid or SelS siRNAs for 30 h. The eNOS expression was tested after 6 h TNF-α treatment. The results are representative of triplicate independent experiments and are presented as mean ± SD, (). versus control; versus empty vector; versus empty vector; versus negative siRNA. CCK-8: cell counting kit-8; NO: nitric oxide; eNOS: endothelial nitric oxide synthase; E-vector: empty vector; Pc-SelS: pcDNA3.1-SelS plasmid; Neg.RNA: negative siRNA.

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