Gene Silencing of PTX3 by siRNA Inhibits Expression of Inflammatory Factors in Aortic Valve Interstitial Cells / 华中科技大学学报(医学版)

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 controlAVICs 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