Heparin Inhibit Expressions Of MUCIN5AC And Activity Of EGFR In Pulmonary Epithelial Cell Line(A549) Stimulated By PMA

Objective To study the influence of heparin on the activity of EGFR and expressions of mucin5AC in pulmonary epithelial cell line(A549).Methods (1) The experiments were performed in culture of PMA induced A549 in vitro .(2)A549 cells divided into 5 groups :the control group, the model group(PMA10), three the intervention groups (G50: 50U/ml concentration of heparin+PMA; G150: 150U/ml of heparin+PMA ;G450: 450U/ml of heparin+PMA).The intervention group,s cell were pretreated with inhibitors(heparin) for 30 min before exposure to stimuli(PMA).(3)The effect of heparin/PMA on epithelial proliferation was assayed using methyl thiazolyl tetrazoliu(MTT) method . The MUC5AC mRNA levels were analyzed by RT-PCR,MUC5AC mucin protein were detected by enzyme linked immunosorbent assay( ELISA) respectively , and phosphorylated epidermal growth factor receptor (p-EGFR)protein production was assayed by western blot.Results (1) The five groups OD detected by MTT were no significant difference (p<0.005). (2) A549 cell was stimulated with PMA and the expressions of mucin5AC was increased(compared with the control group p<0.01), but it was inhibited dose dependence by heparin(compared with the intervention group p<0.005). (3) A549 cell was stimulated with PMA and the activity of EGFR was increased(compared with the control group p<0.01), but it was inhibited by heparin , but there was no significant difference between the model group and the G50 group(P>0.05), there was significant difference between the model group and the G150/450 group(p<0.05,respectively).Conclusions Heparin inhibited expressions of mucin5AC in pulmonary epithelial cell line( A549) stimulated by PMA,it may be by inhibited activity of EGFR. Heparin may be used as a kind of drug against the hypersecretion of mucus.