The role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia induced apoptosis in alveolar epithelial cell A549
10.3969/j.issn.1000-3606.2013.11.017
- VernacularTitle:PKCβ/p66Shc氧化应激通路介导高氧诱导人肺泡上皮细胞凋亡的作用
- Author:
Zhongli CHE
;
Wenbin DONG
;
Qingping LI
;
Xiaoping LEI
;
Lan KANG
;
Lin GUO
;
Xuesong ZHAI
;
Feng CHEN
- Publication Type:Journal Article
- Keywords:
hyperoxia;
protein kinase C beta;
oxidative stress;
alveolar epithelial cells;
apoptosis
- From:
Journal of Clinical Pediatrics
2013;(11):1066-1069
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To explore the role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia-induced apoptosis of alveolar epithelial cells A549. Methods A549 cells were cultured in vitro and divided randomly into control (incubated with 5%CO2), hyperoxia group (exposed to a mixture of 900 ml/L O2 and 50 ml/L CO2 at speed of 3 L/min for 10 mins, then cultured in a closed environment) and LY333531 group (treated with 10μmol/L of PKCβinhibitor LY333531 for 24h then induced with hyperoxia for 10 mins). The cellular morphology was observed under inverted microscope at 12, 24 and 48 h of treatment. The cell apoptosis was detected by lfow cytometry. Expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were detected by immunohistochemistry after 24 h of treatment. Results Comparing to the control group, the cellular morphology of A549 in the hyperoxia group changed to spherical shapes and space between cells increased, the living cell count decreased and suspension cell increased. The living cell count in LY333531 group increased and suspension cell decreased than those in hyperoxia group but not reach the levels of the control group. The apoptosis rate of A549 cells and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 at 24 h were signiifcantly increased in the hyperoxia group than those in the control group, while the apoptosis rate and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were greatly decreased in the LY333531 group than those in the hyperoxia group (all P<0.01). Conclusions The expression of PKCβin A549 cells can be increased by the hyper-oxia induction but reduced by LY333531, and then the expressions of Pin1, p66Shc and p66Shc-Ser36 are reduced. Thus the re-duced apoptosis of A549 cells relieve the cell injury induced by hyperoxia.
