The Role Of CD36 Polyclonal Antibody In M1 And M2macrophage In Response To Ox-LDL In Rats

Objective:To explore the effect of CD36 polyclonal antibody on the cytokines secretion,surface marker expression and foam cell of M1 and M2 macrophages in response to the oxidized low-density lipoprotein in rats.Methods:Monocytes were isolated from peripheral blood of 30 rats given normal diet,and monocytes were induced as initial macrophages with macrophage colony stimulating factor. In vitro, macrophages were differentiated into M1 or M2 macrophages with lipopolysaccharide combined with interferon gamma or interleukin-4 in turn. The different subtype of macrophages were treated with oxidized low-density lipoprotein,CD36 polyclonal antibody prior to the stimulation with oxidized low-density lipoprotein respectively. CD86 on M1 macrophages surface and CD163 on M2 macrophages surface detected with immunofluorescence method, were analyzed with a Image-Pro Plus6.0software for mean fluorescence intensity.The concentration of tumor necrosis factor alpha, interleukin-6 and interleukin-10 in the culture medium of the M1 and M2 macrophages were measured with enzyme linked immunosorbent assay(ELISA) before and after the intervention. The ability of M1 and M2 macrophages to form foam cells was compared before and after the action of CD36 antibody. Using SPSS.16 statistical software to analyze the data, P<0.05 had statistical significance.The independent samples between the two groups were compared with t test, and the Chi-squared test was used for comparing the rate of foam cell formation.Results:(1) The production of interleukin-10 from M1 and M2 macrophages blocked by CD36 polyclonal antibody prior to the stimulation with oxidized low-density lipoprotein were higher than M1 and M2 macrophages stimulated with oxidized low-density lipoprotein, the difference was statistically significant(P < 0.001).(2) The production of tumor necrosis factor alpha, interleukin-6 and the expression of CD86 from M1 macrophages blocked by CD36 polyclonal antibody prior to the stimulation with oxidized low-density lipoprotein were less than those from M1 macrophages stimulated with oxidized low-density lipoprotein, the difference was statistically significant(P <0.05).(3) The production of tumor necrosis factor alpha, interleukin-6 from M2 macrophages blocked by CD36 polyclonal antibody prior to the stimulation with oxidized low-density lipoprotein were less than those of M2 macrophages stimulated with oxidized low-density lipoprotein, then the former CD163 expression was higher than the latter, and the difference was statistically significant(P < 0.001).(4) Blocking CD36 reduced the rate of M1 and M2 macrophages to form foam cells(P < 0.001).Conclusion:CD36 polyclonal antibody inhibited oxidized low-density lipoprotein’s pro-inflammatory effects on M1 and M2 macrophages, and the production of IL-10 from M1 and M2 macrophages increased after the CD36 on two subtypes of macrophages were blocked by CD36 polyclonal antibody. The CD36 polyclonal antibody reduced the formation of foam cells from M1 and M2 macrophages.