Effects of TNF-alpha on MCP-1 and IL-8 mRNA expression in human umbilical vein endothelial cells / 中国实验血液学杂志

ABSTRACT

This study was aimed to explore monocyte chemotactic protein 1 (MCP-1) and interleukin 8 (IL-8) expressions in human umbilical vein endothelial cells (HUVECs) affected by TNF-alpha and its molecular mechanism. RT-PCR assay was used to detect the expression of MCP-1 and IL-8 mRNA in HUVECs at various times following stimulation with TNF-alpha; the nuclear factor-kappa B (NF-kappaB) activation in HUVECs were detected by immunofluorescence. The results showed that after stimulation with NF-kappaB, the expression levels of MCP-1 and IL-8 mRNA in HUVECs increased, and changed along with different time; the expression of IL-8 mRNA reached to peak level at 8 hours and the expression of MCP-1 mRNA reached to peak level at 12 hours; the expression of NF-kappaB p65 protein began increasing at 30 minutes and reached to peak level at 1 hour; then the staining of cytoplasm gradually weakened, while staining of nuclei was enhanced, which indicated significant nuclear translocation of NF-kappaB p65. It is concluded that the TNF-alpha induces expressions of IL-8 mRNA, MCP-1 mRNA and NF-kappaB in HUVECs, and NF-kappaB activities signal pathway may play a role in IL-8 mRNA and MCP-1 mRNA expressions.