STAT6沉默对弓形虫感染NR8383巨噬细胞iNOS、Arg-1基因表达和虫体增殖的影响OA

This study was aimed at studying the effects of Toxoplasma gondii infection on the expression of iNOS and Arg-1 genes in STAT6 silenced NR8383 macrophages.The inhibitory sequence was first designed and synthesized according to the STAT6 gene sequence,PCR amplified,ligated into the GV493 vector plasmid,and transformed into DH5α competent cells.Positive clonal transformants were selected for sequencing and identification.The supernatant was collected after transfection of a lentiviral vector for production of packaged STAT6,to determine the titer.After lentiviral infection,NR8383 rat alveolar macrophages were screened with puromycin to obtain cell lines with stable low STAT6 expression.Real-time PCR and western blotting were conducted to verify STAT6 gene and protein expression.The Toxoplasma gondii RH strain was co-cultured with negative control(NC)and transfected lentiviral group(LV)cells.The infected cells were collected,and the gene and protein expression of factors such as Arg-1 and STAT6 was de-tected.Differential staining analyses were used to assess the proliferation of Toxoplasma gondii in STAT6-silenced cells.Chronic dis-ease was successfully modeled,and stable cell lines were selected.Real-time PCR and western blotting indicated that the gene and protein expression of STAT6 was significantly lower in the LV group than the blank group and the NC group.After Toxoplasma infec-tion,the expression of Arg-1 in the NC group and the LV2 group significantly differed(t=21.65,p<0.001).In conclusion,Toxoplasma gondii regulates macrophage polarization through STAT6,thereby promoting Arg-1 expression,inhibiting STAT6 expression,and at-tenuating Toxoplasma gondii proliferation.