The Effect Of Treg Cells On Polarization Of M1/M2 Macrophages And In Vitro Studies

ObjectiveThe co-culture system of Treg cells and M1/M2 type macrophages was established to observe the effect of Treg cells on the polarization of M1/M2 type macrophages in vitroMethodsUsing age-appropriate C57BL/6 female mice,the adherent method was used to extract the type M1/M2 macrophages from the abdominal cavity,and the spleen magnetic beads were used to separate CD4~+T and induce the differentiation of Treg.Under the condition of M1/M2 macrophages differentiation,Treg was co-cultured with macrophages.The experiment was divided into 6 groups:M,M1,M2,Treg+M,Treg+M1 and Treg+M2 groups.The expressions of F4/80~+i NOS~+M1 macrophages and F4/80~+CD206~+M2 macrophages were detected by flow cytometry(FACS).RT-PCR was used to detect the m RNA expression of IL-10,TGF-β1,arg-1 and i NOS.The expression of IL-10 and TGF-β1 in supernatant was detected by ELISAResultsFACS results showed that compared with M1 group,the expression of F4/80~+i NOS in Treg+M1 group increased.Compared with M2 group,the expression of F4/80~+CD206 in Treg+M2 group was decreased.RT-PCR results showed that,compared with M1 group,the expression of inos m RNA in Treg+M1 group increased,and the expression of arg-1,IL-10 and TGF-β1 was not significant.Compared with M2 group,the expression of arg-1m RNA in Treg+M2 group decreased,and the expression of i NOS IL-10 and TGF-β1was not significant.ELISA results showed that,compared with M1 group,the expression of IL-10 and TGF-β1 in supernatant of Treg+M1 group was decreased,and that of IL-10 and TGF-β1 in supernatant of Treg+M2 group was decreasedConclusionTreg cells can affect the polarization of M1/M2 type macrophages in vitro,that is,the purified Treg cells can promote the differentiation of M1 type macrophages and inhibit the differentiation of M2 type macrophages...