The Study Of The Effect Of USP22 On Vascular Calcification And Related Mechanism

Objective: To investigate the effect of USP22 on osteogenic differentiation and calcification of human aortic vascular smooth muscle cells and its related mechanism.Methods: The calcification model was established with p-diphosphate glycerin,and the alizarin red staining and Western Blot were used to verify the success of the model.The role of estrogen and its receptor in osteogenic differentiation and calcification induced by phosphate was confirmed by calcium quantitative assay.The expression of USP22 was determined by Western Blot and immunofluorescence.The Flag-USP22 plasmid model was constructed,and the effect of USP22 on the osteogenic differentiation and calcification of HASMCs induced by high phosphate was determined by alizarin red staining and Western Blot.Immunofluorescence staining and confocal experiment were used to explore the co-localization of USP22 and ER?,and CO-IP experiment was used to explore the interaction between USP22 and ER?.The effect of USP22 on transcriptional activity of ER? was investigated by using dual-luciferase reporter assay and Western Blot assay.CO-IP and dual-luciferase reporter assay was used to investigate whether the regulation of ER? gene level by USP22 was associated with HDAC1.Results: High phosphate promoted osteogenic differentiation and calcification of HASMCs in a dose-dependent manner,while estrogen inhibited the effect of high phosphate on HASMCs in a concentration-dependent manner.USP22 was highly expressed in high-phosphate-induced HASMCs and promoted high-phosphate-induced HASMCs calcification,which could be inhibited by estrogen.USP22 binds to ER? and HDAC1 to inhibit the transcriptional activity of ER? and promote osteogenic differentiation and calcification of HASMCs.Conclusion: USP22 is highly expressed in calcified HASMCs and promotes osteogenic differentiation and calcification of HASMCs by inhibiting the transcriptional activity of ER? by binding to HDAC1 and ER?.