The Influence Of Blocking Kv1.3 On Macrophage Polarization Induced By Inflammation Cytokines In Atherosclerosis

Background Atherosclerosis (atherosclerosis, AS) has been a more and more serious threat to the human life in world, but also the prevention and treatment of AS is still a global public health puzzle. At present the common belief is that As is a chronic inflammatory disease. In the process of As plaque formation、development and rupture, macrophages play a very important role. Macrophages in the organization can be divided into two different subtype, one type is M1 type (classic activation type), which is a proinflammatory phenotype and can assist Thl cells involved in inflammation and kill the tumor cells. However, the another type is M2 type (alternative activation type), which mainly control tissue repair and remodeling, and promote thr Th2 cells to participate in the immune response. The phenotypic differentiation between Macrophages M1 and M2 can adjust the lymphocyte subsets Th1/Th2 polarization. Studies show that As the existence of Th1/Th2 imbalance and M1/M2 imbalance, thus the regulation of macrophage phenotypic differentiation will help to curb M1 macrophages and Thl lymphocytes mediated proinflammatory response and enhance type M2 macrophages and Th2 lymphocyte mediated anti-inflammatory effects.Voltage-dependent potassium channel on the macrophage cell membrane is one of the important channels which are involved in the regulation of cell function, wherein the potassium channel Kv1.3 is the major subtype of monocyte-macrophage cell Kvl family channels. The self-designed and synthesized antibody against the people E314 peptides of Kv1.3 potassium channels (hKvl.3E314 antibody) can specifically inhibit the Kvl.3 channel on macrophage cell membrane. The study found that blocking the potassium channels on the surface of the macrophages can reduce the secretion of proinflammatory cytokines from the macrophages, on the contrary the overexpression of macrophages Kvl.3 can significantly increase the level of TNF-a from M1 macrophages, and After silencing expression of the macrophages Kv1.3 gene, TNF-a levels were significantly reduced. Thus, Kv1.3 channels for the function changes of macrophages has an important regulatory role.Objective To investigate the effects of blocking Kvl.3 on macrophage polarization induced by inflammation cytokines in the process of occurrence and development of atherosclerosis. To investigate the effects of Kv1.3 channel blockers, in order to provide new methods for the As prevention.Methods The THP-1 monocytes bought from the cell bank of Chinese academy of sciences were given PMA 100ng/ml 72h induced to differentiate into macrophages. The adherent macrophages were seeded in 6-well plates,which were divided into four groups numbered 1,2,3,4:group 1 was joined LPS and IFN-γto incubate cells; group 2 cells were given hKvl.3E314 antibodies; group 3 was given hKvl.3E314 antibodies and LPS and IFN-y in turn forincubation; group 4 cells were given PBS. An inverted phase contrast microscope was used to watch the cell morphology; flow cytometry and qRT-PCR was used to identify macrophage phenotype marker molecules iNOS, Arg-1 and CD206; ELLIS A was used to detect the expression level of IL-10, IL-23, TNF-a in cell supernatants.Results Flow cytometry and qRT-PCR results of four groups showed that LPS and IFN-y synergies can promote macrophage iNOS expression increased(P<0.05), while CD206 and Arg-1 expression decreased(P<0.05). The macrophages induced by hKv1.3E314 antibodies+LPS+IFN-y can promote CD206 and Arg-1 to express highly and iNOS lowly(P<0.05). ELLISA showed LPS+IFN-γ can promote macrophages to secrete IL-23, TNF-a significantly higher (P<0.05), while hKv1.3E314 antibodies+LPS+IFN-γ can promote IL-10 expression levels to significantly increase (P<0.05). When macrophages were only interventioned by hKv1.3E314 antibodies, the results of streaming results, qRT-PCR and ELLISA suggest no significant difference with the PBS intervention group (P> 0.05).Conclusion The Kv1.3 channel is involved in the regulation of macrophage phenotype and we can promote macrophages toward M2 macrophages differentiation which has anti-inflammatory effect by blocking the Kvl.3 channel macrophages.