The Effect Of ADMA On Monocyte Activate Into Macrophage And On Expression Of LOX-1 And MCP-1

Objective:Atherosclerosis (AS) is a chronic, complex vascular inflammatory disease, and the monocytes activate into macrophage is the initial and key event in the development of AS. Asymmetric dimethylarginine (ADMA) is an endogenous nitric oxide synthase (NOS) inhibitor, can mediate inflammatory reaction via induction of NOS uncoupling to increase intracellular oxygen free radicals generation contributing to the process of AS. The role of ADMA on the monocytes activate into macrophage is still not clear. We aimed to explore the influence of ADMA on monocy tes/macrophages chemotaxis by observing the regulation of ADMA on the expression of monocyte chemotactic protein-1 (MCP-1) and lectin-like oxidized low density lipoprotein receptor-1 (LOX-1).Method:(1) Observate the role of ADMA in the expression of MCP-1 in the monocytes:ADMA (20umol/L) or ADMA (20umol/L) plus L-arginine(L-Arg) (1.2mmol/L)were incubated with monocytes for 48h, determination of the level of MCP-1 in concentration medium were measured by ELISA, the mRNA expression of MCP-1 in cell measured by RT-PCR method; (2) Observate the role of ADMA in the expression of LOX-1 in the process of monocytes activate macrophages. First, use the PMA inducted the monocytes activate into macrophages, in this process, ADMA (20umol/L) or ADMA (20umol/L) plus L-Arg (1.2mmol/L) were incubated with monocyte for 48h,Western Blotting analysis of the expression of LOX-1 protein. DAB staining method, the mRNA expression of LOX-1 in cell were measured by RT-PCR method. Statistical evaluation was performed with 16.0 software. Data were expressed as Mean+ SD. Differences among group were compared by ANOVA and were analyzed by q test, P<0.05 was considered statistically significance.Results:(1) Compared with the control group, treatment of monocytes with 20umol/LADMA for 48h, resulted in an increase in the mRNA and protein levels for MCP-1 obviously (P<0.01). (2) Compared with the control group, treatment of monocytes with 20umol/l ADMA+ 1.2mmol/L L-Arg for 48h, the difference of mRNA and protein expression for MCP-1 were insignificance (p>0.05).(3) Compared with the 20umol/L ADMA group, treatment of monocytes with 20umol/L ADMA+1.2mmol/L L-Arg for 48h, resulted in an decrease in the mRNA and protein expression for MCP-1 obviously (P<0.01).(4) Compared with the control group, treatment of monocytes were incubated plus PMA with 20umol/L ADMA for 48h, resulted in an increase in the mRNA (P<0.05) and protein levels obviously (P<0.01) for LOX-1.(5) Compared with the control group, treatment of monocytes were incubated plus PMA with 20umol/l ADMA+1.2mmol/L L-Arg for 48h, resulted in an increase in the mRNA and protein levels for LOX-1 (P<0.05), and treatment with 1.2mmol/L L-Arg for 48h, the difference of the mRNA and protein expression for LOX-1 were insignificance (p>0.05).(6) Compared with the 20umol/L ADMA group, treatment of monocytes were incubated plus PMA with 20umol/L ADMA+1.2mmol/L L-Arg for 48h, resulted in an decrease in the mRNA (P<0.05) and protein levels obviously (P<0.01) for LOX-1, and treatment with 1.2mmol/L L-Arg for 48h, resulted in an decrease in the mRNA (P<0.05) and protein levels obviously (P<0.01) for LOX-1.(7) Compared with the 20umol/L ADMA+1.2mmol/L L-Arg group, treatment of monocytes were incubated plus PMA with 1mmol/L L-Arg for 48h, resulted in an decrease in the mRNA and protein levels for LOX-1 (P<0.05).Conclusion:(1) ADMA can upregulate the expression levels of protein and mRNA for MCP-1 and LOX-1, L-Arg can antagonize the effects of ADMA prove that ADMA may promote monocyte activate into macrophage cell.(2) ADMA could promote monocyte activate into macrophage cell prove that ADMA is a risk factor in the AS.