Polarization Of THP-1-derived Macrophages Induced By Ox-LDL Mediated By β₂ Adrenergic Receptor

Background:Atherosclerosis is the basis of a variety of vascular diseases,a serious threat to human health.Objective:To observe the effect ofβ₂adrenergic receptor,β₂adrenergic receptor blocker and stimulant on the polarization of ox-LDL induced THP-1derived macrophages.Methods:THP-1 monocytes were induced to transform into macrophages with the concentration of PMA at 100 ng/m L.THP-1 cells were cultured in good condition,and M0macrophages were obtained after 48h induction with 100 ng/m L PMA concentration.First,different concentrations of ox-LDL were treated for 24 h.After treatment,cell activity was detected by CCK8to determine the safety of the drug.Then,the next experiment was carried out.According to the previous MTT drug cytotoxicity test,β₂adrenergic receptor blocker（ICI118 551）10μmol/L andβ₂adrenergic receptor agonist（salbutamol）10μmol/L were used as the subsequent experimental intervention concentrations.The cells were cultured to a good state,treated with ox-LDL,ICI118551,and salbutamol at different concentrations for 1 h,and then treated with ox-LDL for 48 h.The cells were collected and treated by oil red O staining to determine the degree of cell foaming after ox-LDL phagocytosis by THP-1-derived macrophages,and the foam cell model was constructed.After the successful construction of macrophage foaming model,ICI118 551,salbutamol,and salbutamol+ICI118 551 were treated with ICI118 551,respectively.The blank control group,ox-LDL group,ox-LDL+ICI118 551 group,ox-LDL+salbutamol group,ox-LDL+salbutamol+ICI118 551 group were set.The expression levels of CD206,Arg-1,TNF-αand i NOS were detected by q PCR.ELISA method to detect ICI118 551,salbutamol,ICI118 551+salbutamol after intervention of ox-LDL induced THP-1 source sex macrophages release of IL-10 and streaming apoptosis apoptosis detection to verify that the ox-LDL on the degree of polarization M0 macrophages and observe theβ₂adrenergic receptor mediated,β₂adrenergic receptor blockers and stimulant for ox-LDL induced THP-1 source sex M1,M2 macrophages degree of polarization transformation.Results:As shown under the microscope,THP-1 monocytes could be successfully induced to differentiate into M0 macrophages after 48h treatment with 100ng/ml PMA.Oil red O staining showed that THP-1derived macrophages foaming could be successfully induced when the concentration was 100μg/m L ox-LDL.100μg/m L of ox-LDL was selected as the treatment concentration for the next experiment.Flow cytometry analysis showed that different drug treatments had significant differences in the apoptosis of M0 macrophages.Salbutamol could enhance the occurrence of apoptosis after the macrophage phagocytosis of ox-LDL,ICI118551 could weaken the occurrence of apoptosis after the macrophage phagocytosis of ox-LDL.ELISA assay showed thatβ-blocker ICI118551 effectively inhibited the release of IL-10 compared with blank control group,andβ₂agonist effectively promoted the release of IL-10.q PCR assay showed that all treatments effectively promoted the upregulation of i NOS gene and inhibited the expression of TNF-αgene.ox-LDL+IC118551 group significantly inhibited the upregulation of CD206 gene,but the difference level was not significant in other groups.For Arg-1 gene,ox-LDL+IC118551 group significantly inhibited the expression of Arg-1 gene,ox-LDL+salbutamol group effectively promoted the expression of Arg-1 gene.Conclusion:β₂receptor blockers can promote the polarization of macrophages to M1 and inhibit the polarization of macrophages to M2.β₂receptor stimulant can inhibit the polarization of macrophages to M1 and promote the polarization of macrophages to M2.β₂receptor blockers andβ₂receptor stimulants may induce apoptosis of macrophages throughβ₂-ARS.