Effect of vascular injury on the expression of polymerase Ⅰ and transcript release factor in vascular smooth muscle and its mechanism

doi:10.3969/j.issn.1006-7795.2017.04.012

Abstract

Abstract: Objective To investigate the role of polymerase Ⅰ and transcript release factor or Cavin-1 in vascular restenosis after vascular injury and its molecular mechanism. Methods 200-220 g Sprague Dawley(SD)male rats were randomly divided into sham and injury groups with injury groups establishing balloon injury models. Cellular content from carotid taken after the sham and balloon injuries were shown by hematoxylin and eosin staining. Expression Cavin-1 protein from above groups were determined by Western blotting and immunofluorescence. The Cavin-1 mRNA from above groups was detected by real-time RT-PCR, respectively. Immunochemical staining, Western blotting and Co-immunoprecipitation revealed ubiquitinated protein in carotid artery from sham and injury groups. Rat aortic smooth muscle cells were divided into: ①control group(CTRL);②CHX group rats were pretreated with cycloheximide (CHX, 25μmol/L) for 1 h;③CHX+MG group rats were pretreated with CHX (25μmol/L) for 1 h followed by treatment with MG 132(10 μmol/L)for additional 24 hours.④CHX+CQ group rats were pretreated with CHX (25μmol/L) for 1 h followed by treatment with chloroquin(CQ,50 μmol/L)for additional 24 hours.Expression Cavin-1 protein from above groups were determined by Western blotting. Results As for animal models, the carotid vascular wall became thicker after the balloon injury and the expression of Cavin-1 protein was significantly decreased(P<0.05) while real-time RT-PCR showed no significant difference of Cavin-1 mRNA between sham and injury groups. Ubiquitinated protein levels were higher in the injured carotid compared with sham group(P<0.05) As for cell experiment, expression of Cavin-1 was decreased significantly from CHX group compared with CTRL(P<0.05).CHX+MG group can reverse the above effects (P<0.05). Conclusion Expression of Cavin-1 protein from balloon injury carotid arteries were reduced, and its mechanism may relate to up-regulated ubiquitination degradation pathway.

Key words: vascular injury, smooth muscle cell, ubiquitination, vascular restenosis

CLC Number:

Chen Xueying, Yao Ye, Zhou Gaoshi, Tao Shiwan, Chen Zhaoyu, Tan Zhi. Effect of vascular injury on the expression of polymerase Ⅰ and transcript release factor in vascular smooth muscle and its mechanism[J]. Journal of Capital Medical University, 2017, 38(4): 546-552.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2017.04.012

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2017/V38/I4/546

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