This study aimed to explore the regulation mechanism of DNA methylation editing of nitric oxide synthase(NOS)2 on Myoglobin,and study its effect on the expression level of neuronal nitric oxide synthase(nNOS)and endothelial nitric oxide synthase(eNOS).In this experiment,DNA methylation editing technology was used to methylate NOS2 in porcine kidney cell line PK-15 cells,and the DNA methylation level of the NOS2 gene promoter,the expression level of NOS2,the concentration of intracellular nitric oxide,as well as the expression level of myofibrillar-and myoglobin-related genes were detected in porcine kidney cells.The results revealed that the average DNA methylation level of the first 500 bp region of the NOS2 gene promoter was reduced,but the difference was not significant,while the methylation level of the CpG site near the gRNA2 binding site was increased.In addition,after DNA methylation editing of NOS2 gene,NOS2 gene expression and its iNOS protein expression were significantly reduced.With the decrease of NOS2 expression,nNOS and eNOS expression increased significantly,the expression level of MYHC-Ⅱ b and MYHC-Ⅱ x decreased significantly,and the expression of Myoglobin protein decreased significantly.In conclusion,the abnormal expression of the NOS 2 gene can affect the generation of NO in cells,as well as the expression of NOS 1 and NOS 3,and the expression of genes related to muscle development.
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