The role  of ZFP580 in the regulation of rat VSMCs migration by  ATRA and its mechanism

doi:10.12047/j.cjap.5806.2020.010

Abstract

Abstract: Objective:To investigate the probable roles of the novel C2H2 zinc finger transcription factor ZFP580 on all-transretinoic acid (ATRA)-regulated VSMCs migration and underlying mechanisms. Methods: Rat aortic VSMCs were isolated, cultured and identified. VSMCs were treated with ATRA at the concentrations of 0, 5, 10 or 20 μmol/L for 24 hours. The migration ability of VSMCs was observed in each group and compared with control group which was treated by 0 μmol/L ATRA. The mRNA and protein expression levels of ZFP580 were detected by QPCR and Western blot. ZFP580 protein expression in VSMCs was detected under ATRA stimulation when ERK inhibitor PD98059 was used to inhibit the protein expression of ERK. Adenovirus transfection technology was used to obtain VSMCs with overexpression or low expression of ZFP580, and QPCR and Western blot were used to detect the mRNA and protein levels of MMP-2, MMP-9 and ZFP580.Results: On the 10th day of VSMCs culture, immunofluorescence showed that SM22 alpha antibody, as a specific marker of smooth muscle cells, was positive. Compared to the control group, VSMCs migration was reduced by 32%, 43%, and 59% in the group of 5, 10, and 20 μmol/L ATRA pretreatment. Compared with the control group, VSMCs treated by 20 μmol/L ATRA reduced the cell migration by 49%, 36% and 22% at 24, 48 and 72 h. The mRNA and protein expression levels of ZFP580 were increased with the increase of ATRA stimulation solubility and the extension of stimulation time. ERK was increased significantly after 15 min of ATRA stimulation. Pretreatment with ERK inhibitor PD98059 (20 μmol/L) inhibited the expression of ERK protein and reduced the expression of ATRA-induced ZFP580 protein. Overexpression of ZFP580 inhibited the expressions of MMP-2 and MMP-9, whereas down-expression of ZFP580 promoted the expressions of MMP-2 and MMP-9.Conclusion: ATRA increased the expression of ZFP580 through the ERK signaling pathway, while ZFP580 was involved in ATRA's inhibition of VSMCs migration by affecting the expression of downstream MMP-2 and MMP-9.

Key words: ZFP580, vascular smooth muscle cells, all-trans retinoic acid, migration, mouse

WU Guo-dong, CHEN Kai, ZHANG Jing-jing, WEI Shu-ping, ZHANG Mei, ZHANG Wen-cheng. The role of ZFP580 in the regulation of rat VSMCs migration by ATRA and its mechanism[J]. CJAP, 2020, 36(1): 45-50.

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The role of ZFP580 in the regulation of rat VSMCs migration by ATRA and its mechanism

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