| Chapter 1. Pulmonary vascular endothelial cells apoptosis and the expression of NF-κB in COPD patientsObjective:To investigate apoptosis and the expression of nuclear factor-kappa B(NF-κB) in the pulmonary vascular endothelial cells from patients with chronic obstructive pulmonary disease.Methods:According to the latest COPD diagnostic criteria and WHO standards for smoking-related investigation,19 patients underwent pneumonectomy were divided into 3 groups:group A:non-smokers without COPD; group B:smokers without COPD; group C:smokers with COPD. The HE staining was used to evaluate the pathology changes of lung tissues. The apoptosis of pulmonary vascular endothelial cells was ascertained by using the methods of TdT-mediated dUTP nick end labeling(TUNEL). Immunohistochemistry was used to investigate the expression of nuclear NF-κB in the pulmonary vascular endothelial cells.Results:The apoptotic rate of pulmonary vascular endothelia cells in group C [(11.87±0.79)%] was higher than that in group B [(8.80±0.63)%] and group A [(4.84±0.45)%],differences between every two groups were of significance(P<0.05).The expression rate of NF-κB p65 in group A was (3.72±0.33)%, in group B was(15.92±1.13)%, and in group C was(37.23±4.75)%.Each variant increased in turn and differences between every two group were significant(all P<0.05).Conclusions:Increased apoptosis and expression of NF-κB exists in pulmonary vascular endothelial cells in the smokers with COPD and the smokers without COPD. Cigarette smoking may be related with increases of pulmonary vascular endothelial cell apoptosis and NF-κB expression. Chapter 2. Pulmonary vascular endothelial cells apoptosis and the expression of NF-κB in COPD mice modelsObjective:To investigate the pulmonary vascular endothelial cells apoptosis and the expression of nuclear factor-kappa B(NF-κB) in the pulmonary vascular endothelial cells from mice with chronic obstructive pulmonary disease.Methods:16 C57BL/6J male mice were divided into control group(8 mice) and COPD group(8 mice), the COPD model was established by exposing the mice to cigarettes smoke. The HE staining was used to evaluate the pathology of lung tissues, the methods of TUNEL was uesed to ascertain apoptosis of pulmonary vascular endothelial cells, and immunohistochemistry was used to investigate the expression of nuclear NF-κB in the pulmonary vascular endothelial cells.Results:The apoptotic rate of pulmonary vascular endothelia cells in COPD mice group [(15.91±1.15)%] was higher than that in control group [(3.39±0.64)%],differences between the two groups were of significance(P<0.05).The expression rate of NF-κB p65 in COPD mice group was(35.30±4.20)%,in control group was(6.37±0.54)%.The variant increased in the COPD mice group and differences between the two groups were significant(all P<0.05). Conclusions:Increased apoptosis and expression of NF-κB existsin the pulmonary vascular endothelial cells is increased in the cigarette smoking COPD mice group. Chapter 3. The effect of NF-κB on the endothelial cell apoptosis induced by cigarette smoking extractObjective:To investigate the function of NF-κB in CSE-induced HUVEC apoptosis.Methods:Human umbilical vascular endothelial cells(HUVECs) were cultured in vitro and given different interfered:In part one, HUVEC were exposed at 0%,2.5%,5%,10% and 20% CSE for 12h; In part two, HUVEC were exposed at 5%CSE for 0.5h,2h,4h,8h and 12h; In part three, after being interfered with 100μmol/l pyrrolidine dithiocarbamate(PDTC) in 0.5h, HUVECs were interfered with 5% CSE for 1h and 2h. The translocation rate of NF-κB was tested by immunohistochemistry, the apoptosis rate of HUVEC was evaluated by using the flowcytometry with Annexin V/PI double staining.Results:The translocation rate of NF-κB p65 increased in the HUCEC in CSE-dose dependent manner when CSE concentration was less the 5%. And in 5% CSE interfering group, it reached the maximum[(18.13±1.37)%], differences between the other groups were of significance (P<0.05). The translocation rate of NF-κB p65 was maximum[(93.80±3.12)%] when HUVEC were exposed to 5% CSE for 0.5h, and it decreased with the exposure time was prolonged. The apoptosis rate of the HUVEC increased in a dose-dependent manner, and in 10% CSE interfere group it was the maximum[(45.19±4.91)%], differences between the other groups were of significance(P<0.05).The apoptosis rate increased in a time-dependent manner when HUVEC were exposed to 5% CSE. After being interfered PDTC, the translocation rate of NF-κB p65 induced by 5% CSE in HUVEC in 1h and 2h interfering group were(5.10±0.55)%,(4.51±0.85)%, were much lower than the non-pretreated groups[(31.93±2.94)%],[(25.32±3.54)%] respecting significant difference (P<0.05). After being interfered PDTC, the apoptosis rate in 1h and 2h interfering groups[(11.20±1.11)%],[(38.83±3.43)%] were much higher than the non-pretreated groups[(6.53±0.80)%],[(28.87±3.53)%], respecting significant difference (P<0.05).Conclusions:Cigarette smoking extract induces endothelial cell apoptosis in concentration-and time-dependent effects. The nuclear translocation of NF-κB induced by CSE in HUVEC happens in 0.5h. NF-κB may play a protective role in the development of CSE induced apoptosis of endothelial cells.
|
PDF Full Text Request