Effects of Hcy and lncRNA ANRIL on atherosclerotic inflammatory response / 中国医师杂志

ABSTRACT

Objective:To study the relationship between the level of serum homocysteine (Hcy) and the antisense non coding gene (ANRIL) of long chain non coding RNA (lncRNA) cell cycle dependent kinase inhibitor 2B gene, and the effect on Atherosclerosis inflammation, that is, the expression of interleukin-10 (IL-10) and Monocyte chemoattractant protein-1 (MCP-1) in Human umbilical vein endothelial cell (HUVEC).Methods:HUVEC was cultured in vitro and cells were treated with different concentration gradients (blank control group, 0.5, 1.0, 2.0, 5.0 mmol/L) of Hcy. The expression level of lncRNA ANRIL was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of MCP-1 and IL-10. LipoFilter transfection reagents were used to transfect shANRIL and shNC into different cells, respectively. In the above experiment, the optimal Hcy concentration (5.0 mmol/L) was selected for intervention for 24 hours. Western blot was used to detect the protein expression levels of MCP-1 and IL-10.Results:After 24 hours of intervention with different concentrations of Hcy in HUVEC, Hcy significantly damaged endothelial cells, and the higher the Hcy concentration, the more severe the cell damage. Compared with the blank control group, the Hcy intervention group showed an increase in lncRNA ANRIL and MCP-1, while IL-10 decreased (all P<0.05); As the concentration of Hcy intervention increases, IL-10 decreases, while lncRNA ANRIL and MCP-1 increased (all P<0.05). Compared with the blank control group, the Hcy group, the shNC+ Hcy group, and the shANRIL+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). Compared with the shANRIL+ Hcy group, the Hcy group and the shNC+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). There was no statistically significant difference in the expression levels of IL-10 protein and MCP-1 protein between the shNC+ Hcy group and the Hcy group (all P>0.05). Conclusions:Hcy upregulates MCP-1 expression and downregulates IL-10 expression by promoting lncRNA ANRIL expression. Thus, it can promote cellular inflammatory reaction and participate in Atherosclerosis.