Gene Silencing of PTX3 by siRNA Inhibits Expression of Inflammatory Factors in Aortic Valve Interstitial Cells
10.3870/j.issn.1672-0741.2019.05.001
- VernacularTitle:靶向siRNA沉默长正五聚蛋白3对主动脉瓣膜间质细胞表达炎症细胞因子的影响
- Author:
Mei Li
1
;
Fei Li
;
Wei Su
Author Information
1. Department of Cardiovascular Surgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China
- Publication Type:Journal Article
- Keywords:
calcific aortic valve disease;
pentraxin 3;
lipopolysaccharide;
interleukin-6;
interleukin-8;
monocyte chemoattractant protein-1
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2019;48(5):497-502
- CountryChina
- Language:Chinese
-
Abstract:
Objective:
To explore the role of lipopolysaccharide（LPS）in the pathogenesis of calcific aortic valve disease（CAVD）by detecting the expression of inflammatory factors in aortic valve interstitial cells（AVICs），and the expression of interleukin-6（IL-6），interleukin-8（IL-8），monocyte chemoattractant protein-1（MCP-1）after silencing pentraxin 3（PTX3）gene，to test the effect of PTX3 on the pathophysiological process of CAVD.
Methods:We obtained aortic valves for immunohistochemistry staining from 12 patients with CAVD，and from 12 patients without aortic valve disease receiving heart transplant operation as controlAVICs were cultured in vitro，different concentrations of LPS（0，50，100，200 ng/mL）treated cells for 24 h，and then the expression levels of IL-6，IL-8 and MCP-1 were analyzed by real-time polymerase chain reaction（real-time PCR）. After AVICs were transfected with PTX3 siRNA for 48 h to knockdown the expression of PTX3 protein，PTX3 was tested by Western blotting. The levels of IL-6，IL-8 and MCP-1 were detected by real-time PCR 24 h after treatment with LPS（100 ng/mL）in AVICs with PTX3 siRNA transfection.
Results:The calcific aortic valves significantly expressed PTX3 as compared with control.LPS dose-dependently increased IL-6，IL-8 and MCP-1 in AVICs. Compared with control groups，100 ng/mL LPS significantly increased IL-6，IL-8 and MCP-1 mRNA（P<0.05，P<0.01）. PTX3 siRNA markedly decreased the levels of PTX3 protein compared with control groups（P<0.01）. Levels of IL-6，IL-8 and MCP-1 were significantly reduced in LPS plus PTX3 siRNA group compared with controls（all P<0.05）.
Conclusion:The calcific aortic valves have a higher level of PTX3 than control valves.LPS stimulates the expression of IL-6，IL-8 and MCP-1 in AVICs，but silencing PTX3 gene significantly inhibits LPS-stimulated expression of IL-6，IL-8 and MCP-1. PTX3 may play a role in CAVD pathogenesis by regulating expression of inflammatory factors
