The Role And Mechanism Of PGAM1 In Pulmonary Hypertension

Phosphoglycerate mutant enzyme 1(PGAM1),as a glycolytic enzyme,is capable of converting 3-phosphoglycerate(3-PG)to 2-phosphoglycerate(2-PG).Studies have shown that PGAM1 plays important roles in regulating a variety of biological events such as energy production and nucleotide biosynthesis.Studies have shown that PGAM1 is highly expressed in various cancers,including lung cancer,osteosarcoma,and pancreatic cancer,where it functions in promoting cell proliferation,migration,and invasion.However,the role of PGAM1 in pulmonary vascular disease has not been reported.In order to explore the roles of PGAM1 in pulmonary hypertension,this research explored the expression of PGAM1 in the PH animal model,and then focused on the effect of PGAM1 on the proliferation of pulmonary artery smooth muscle cells and on the Endothelial-Mesenchymal Transformation of pulmonary vascular endothelial cells,and revealed the regulatory effect of HIF on PGAM1.The following results were obtained:(1)Expression of PGAM1 in PH animal models: The protein and m RNA expression of PGAM1 were analyzed by Western Blot and q-PCR in lung tissues of HPH,MCT-PH and SUHX-PH animal models.The distribution of PGAM1 in lung tissues was detected by immunofluorescence.The results showed that the protein and m RNA expression levels of PGAM1 in PH animal model were higher than those in control group.(2)The effect of PGAM1 on the proliferation of pulmonary artery smooth muscle cells :The influence of PGAM1 si-RNA or PGMI-004A(PGAM1 inhibitor)were detected by Western Blot and Ed U on the regulation of pulmonary artery smooth muscle cell proliferation.The results show that the deletion of PGAM1 significantly inhibit the proliferation of pulmonary artery smooth muscle cell both under normoxia and hypoxia.(3)The effect of PGAM1 on endothelial to mesenchymal transformation of pulmonary vascular endothelial cells: The influence of PGAM1 deletion by si-RNA or PGMI-004A(PGAM1 inhibitor)were detected by Western Blot and Immunofluorescence on the Endothelial-Mesenchymal Transformation of pulmonary vascular endothelial cells induced by TGF-β.The results show that inhibition of PGAM1 could effectively alleviate the EndothelialMesenchymal Transformation of pulmonary vascular endothelial cells.(4)The regulatory effect of HIF on PGAM1: The expression of PGAM1 were detected by Western Blot and q-PCR in pulmonary artery smooth muscle cells and pulmonary artery endothelial cells after treatment with DMOG.Double luciferase report assay was performed to determine the direct effect of HIF to PGAM1.The results show that the activation of HIF could increase the expression of PGAM1 under normoxia,and HIF can bind to Hypoxia response element of PGAM1 promoter region which plays a role in transcriptional regulation.