Effects of mechanical stretch on expression of pentraxin-3 mRNA and protein in A549 cells
- VernacularTitle:机械牵张对A549细胞穿透素-3表达的影响
- Author:
Huaqing SHU
;
Qingping WU
;
Lei YANG
;
Hong ZHANG
;
Shanglong YAO
- Publication Type:Journal Article
- Keywords:
Respiration,artificial;
Epithelial cells;
Pulmonary alveoli;
C-reactive protein
- From:
Chinese Journal of Anesthesiology
2008;28(11):1023-1025
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the effects of mechanical stretch on pentraxin-3(PTX-3)mRNA and protein expression in human alveolar epithelial cells (A549 cells).Methods The human lung epithelial adenocarcinoma cells A549(A549 cells)were purchased from cell biology laboratory,Tongji Medical College,Huazhong University of Science and Technology.The cultured A549 cells were inoculated on collagen Ⅰ BioFlex plates and divided into 5 groups(n=3 wells each):group Ⅰ normal control;groupⅡsham mechanical stretch;group Ⅲ mechanical stretch;groupⅣsiRNA and groupⅤ siRNA+mechanical stretch.In group Ⅲ the cells underwent square cyclic mechanical stretch for 4 h using the Flexercell Systcm.In group Ⅳ the cells were transfected with chemosynthetic PTX-3 specific siRNA by RNAi technique.In group Ⅴ at 24 h after being transfected with PTX-3 siRNA the cells underwent mechanical stretch for 4 h.In groupⅡ mechanical stretch of the cells were prevented by Flexstep.The expression of PTX-3 mRNA in the cells was detected by real-time PCR and the expression of PTX-3 protein in the culture media was determined by Western blotting.Apoptosis of the cells was measured bv flow cytometry(Beeten-Dickinson,USA).Results PTX-3 mRNA and protein expression was signlficantly up-regulated by mechanical stretch in groupⅢand decreased by transfection with siRNA in group Ⅳand Ⅴ as compared with group Ⅰ andⅡ(P＜0.05 or 0.01).The apoptosis ratio was significantly higher in group Ⅲ and Ⅴ than in groupⅡ and was significantly lower in group Ⅴ thanin group Ⅲ(P＜0.01).Conclusion Mechanical stretch can up-regulate PTX-3 mRNA expression in A549 cells.
