Effect of PGE2 on secretion function and apoptosis of airway smooth muscle cells

Abstract

Objective Airway remodeling is closely related to migration,proliferation,secretion function of airway smooth muscle cells ( ASMCs ). Prostaglandin E2 (PGE2) have a regulatory role in these functions.To culture with different concentration gradient of PGE2 and ASMCs in vitro and to study ASMCs secretion functions of interleukin 4 (IL -4) and interleukin 10 (IL -10) and Interferon-γ (IFN- γ),and apoptosis of ASMCs,this is favorable to understand ASMCs immune regulation mechanism in asthma,and the regulatiun role of PGE2.Methods The rat airway smooth muscle cells and the final concentration of 10 -9～ 10 -6 PGE2 were cultured for 24 hours,divided into 6 groups,and 4 concentration gradient. Flow cytometric were used determination of apoptosis with Annexin V-EGFP/PI double staining apoptosis detection kit,United States Bio-Rad 680 enzyme mark instrument were used detection of its concentration with IL- 10,IL- 4,IFN- γ ELISA kit.4 complex holes into 1 groups,the experiment was repeaied 2 times,average data for 8 holes.Results There were no significant changes in ASMCs cell apoptosis rate after application of the different concentrations of PGE2 compared with the control group.But interleukin secretion and PGE2 has significant dose-effect relationship.When PGE2 concentration were decreased,the secretion of IL- 4 and IL- 10 decreased significantly,but the IFN- γ secretion was increased.(Control,negative,10- 9 PGE2,10- 8 PGE2,10 -7 PGE2,10 -6 PCE2 group,IL- 4 was respectively 6.77 ±1.58,9.24 ± 2.45,38.17 ±11.33,29.27 ±11.12,26.05 ±9.49,21.11 ± 7.21;IL-10 was 4.8 ± 1.06,6.35±2.11,71.89 ±12.03,43.68 ±11.25,28.89 ±8.75,24.31 ± 6.67; 2.17 ± 1.97,IFN-γ 3.25± 1.48,10.63 ± 4.75,13.4 ± 5.57,15.47 ± 6.16,19.54 ± 6.35).Conclusions PGE2 can regulate the secretion of ASMCs cytokines,increased with concentration gradient can promote Th1 cytokines IFN-γ increased secretion,while inhibition of type Th2 cytokine secretion of IL-10,IL-4,but no significant effect on the apoptosis of ASMCs. We think the key role of the normal state of PGE2 secretion were related to maintaining Th1/Th2 balance in the airway. Key words: Airway smooth muscle cells ; Prostaglandin E2 ; Interleukin; Interferon-γ; Apoptosis