The Effects Of Heme Oxygenase-1 On Apoptosis Of Primary Culture Type â…¡ Alveolar Epithelium Cells In Rats

Objective:To investigate the influence of heme oxygenase-1 on apoptosis of typeⅡalveolar epithelial cells (AEC-Ⅱ) of oxidative damage induced by hygrogen peroxide (H2O2) and to observe the mitochondrial damage.Methods:Twenty-six healthy male SD rats were subjected to anesthesia, tracheal intubation and open-chest to take lung, purification, the AEC-Ⅱwere cultured by 1 X 106/ml in cell culture plate, the primary culture lasted for 24 h, then randomly divided into four groups:normal group, H2O2 group, HO-1 group, inhibit group (zinc original porphyrin IX). H2O2 (0.5mmol/L) were used to stimulate AEC-Ⅱto establish acute lung injury cell model.Using HO-1 (0.2 mmol/L) or zinc original porphyrinⅨ(20μmol/L) outside the cells for direct intervention. Electron microscopy identificated the AEC-Ⅱ;Fluorescence microscope,streaming instrument detected the cells apoptosis;streaming instrument detected the cells mitochondria membrane potential;Western blotting detected mitochondria related proteins Bcl-2,p53 expression changes.Results:By electron microscopy to discover cells characteristic structure:surface different length,different thicknesses microvilli, cytoplasm ranging in different size,number and mature stage eosinophils osmium tetroxide sex shelf vorkommen. Fluorescence microscope detection:compared with normal group, H2O2,HO-1 inhibit groups were visible in mass of green and red fluorescent apoptotic cells; HO-1 group's fluorescence cells were obviously reduced. Flow cytometric analysis: compared with normal group, H2O2,HO-1 inhibit group apoptosis rate were increased (P<0.05);with the extension of H2O2,the cell apoptosis rate increased; Mitochondrial membrane potential of depolarization proportion were reduced (P< 0.05).Compared with H2O2 group, HO-1 group AEC-Ⅱcell apoptosis rate reduced significantly (P<0.05); Mitochondrial membrane potential of depolarization proportion increased (P<0.05). Western blotting:compared with normal group, HO-1 group's Bcl-2 expression clearly enhanced, p53 expression is abated in statistically significant difference(P< 0.05). Compared with HO-1 group, H2O2,HO-1 inhibit group's Bcl-2 expression weakened and p53 expression clearly enhanced in statistically significant difference (P< 0.05).Conclusions:HO-1 reduce H2O2 oxidative damage on AEC-Ⅱcell apoptosis rate, and stable mitochondria membrane potential and regulate the mitochondrial related proteins expression of Bcl-2,p53 which possibly participate HO-l's antiapoptosis.