ABSTRACT
AIM:To investigate whether cigarette smoke(CS)promotes the expression of endoplasmic reticu-lum-associated apoptosis protein CCAAT/enhancer-binding protein homologous protein(CHOP)in rat lung tissues. METHODS:Adult male Wistar rats(n=40)were randomly divided into 4 groups with 10 rats in each group: control group,CS-2 group(exposed to CS for 2 months),CS-4 group(exposed to CS for 4 months)and ex-smoking(Ex-S)group (exposed to CS for 4 months and then quit smoking for 1 month).The percentage of forced expiratory volume in 0.3 second to forced vital capacity(FEV0.3/FVC)and peak expiratory flow(PEF)were measured.TUNEL assay was used to detect the apoptotic cells.In situ hybridization and RT-PCR were used to determine the mRNA expression of CHOP.The methods of immunohistochemistry and Western blot were used to determine the protein expression of CHOP.Western blot was also used to determine the protein levels of protein kinase R-like endoplasmic reticulum kinase(PERK),p-PERK,eukaryotic initiation factor(eIF)2αand p-eIF2α.RESULTS:The pulmonary function greatly decreased in the rats exposed to CS for 2 months in comparison with control group(P<0.05),markedly decreased in the rats exposed to CS for 4 months as com-pared with the rats after exposure to CS for 2 months(P<0.05),and was improved little in ex-smoking rats(P>0.05). The structural destruction of the lung was observed in the rats exposed to CS for 2 months,and more obvious changes were found in the rats exposed to CS for 4 months.However,the structural destruction of the lung remained obvious in ex-smok-ing rats.The apoptotic cells were markedly increased in the rats exposed to CS for 2 months and were even more in the rats exposed to CS for 4 months.The apoptotic cells were alveolar epithelial cell I(ACE I),ACE II,vascular endothelial cells and bronchial epithelial cells.The protein levels of p-PERK,p-eIF2αand CHOP were remarkably increased in the rats af-ter exposure to CS for 2 months compared with the control rats(P<0.05),significantly elevated in the rats exposed to CS for 4 months compared with the rats exposed to CS for 2 months(P<0.05),and slightly decreased in ex-smoking rats in comparison with the rats after exposure to CS for 4 months(P>0.05).The total protein levels of PERK and eIF2αdid not change between the control rats and those exposed to CS.CONCLUSION: CS promotes the development of chronic ob-structive pulmonary disease(COPD)by inducing the expression of endoplasmic reticulum-associated apoptosis protein CHOP via PERK/eIF2α/CHOP signaling pathway.
ABSTRACT
<p><b>OBJECTIVE</b>To study the expression of lung Krüppel-like transcription factor (KLF2/LKLF) in lung tissues of rats with chronic obstructive pulmonary disease (COPD) and the relationship between KLF2 and NF-E2-related factor 2 (Nrf2), and make further explore the effects of KLF2 on the expression of gamma-glutamylcysteine synthetase (gamma-GCS).</p><p><b>METHODS</b>Twenty-two male SD rats were randomly divided into a COPD group (n = 10) and a normal control group (n = 11). The rat model of COPD established by cigarette smoking and intratracheal instillation of lipopolysaccharide (LPS), and lung tissues were obtained. The expressions of KLF2, Nrf2, gamma-GCS mRNA and protein in lung tissues were measured by immunohistochemistry (IHC), Western blot, in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR). To explore the relationship between KLF2 and Nrf2 protein,we utilize the method of co-immunoprecipitation (CO-IP).</p><p><b>RESULTS</b>IHC and Western blot showed that protein expressions of KLF2, Nrf2, gamma-GCS were higher in the lung tissues from rats with COPD than those in the control groups (all P < 0.05). The levels of KLF2, gamma-GCS mRNA were markedly increased in the COPD group (all P < 0.01) while Nrf2 mRNA expression in COPD group had no significant difference with that in control group ( P > 0.05). CO-IP result showed that KLF2 were obviously present in immunoprecipitates of Nrf2 (P < 0.01) . Linear correlation analysis showed that the level of KLF2 protein was positively correlated with the level of Nrf2 protein (P < 0.05), and KLF2, Nrf2 proteins were positively correlated with gamma-GCS mRNA and protein (all P < 0.05).</p><p><b>CONCLUSION</b>The expression of KLF2 is significantly up-regulated in COPD, which maybe up-regulate gamma-GCS mRNA expression by increasing Nrf2 expression and nuclear translocation against oxidative stress.</p>