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)
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
; (6): 497-502, 2019.
Article
в Zh
| WPRIM
| ID: wpr-792845
Ответственная библиотека:
WPRO
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
Полный текст:
1
База данных:
WPRIM
Язык:
Zh
Журнал:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
Год:
2019
Тип:
Article