Objective To explore changes of 3’-glutamylcysteine synthetase( γ-GCS)and reduced glutathione(GSH)in patients with bronchial asthma．Methods Ten patients with acute asthma were enrolled and treated for six weeks according to guideline recommendations．Levels of -GCS，GSH and malondialdehyde(MDA)in total cells in induced sputum and GSH，MDA，reactive oxygen(ROS)in selum were measured and compared before and after therapy．Ten healthy volunteers were as normal contro1．Meanwhile，the pulmonary function(FEVl％pred)was measured and asthmatic symptoms were quantified using Hogg’s way．Results A．In serum and sputum of the asthma patients，GSH were lower and MDA were higher before treatment than those of the control(Plt;0．01)．And -GCS in induced sputumwere higher before treatment than those of contro1．B．After treated for six weeks．levels of GSH in serum and sputum of the asthma patients increased copmpared to baseline(all Plt;0．01)，but were still lower than that of control(Plt;0．05)．Activities of MDA in serum and sputum and -GCS in sputum were elevated compared to baseline(Plt;0．01)，but still higher than that of control(all Plt;0．05)．C．Levels of GSH in serum of all patients were correlated negatively witll asthmatic symptom scores and levels of MDA and ROS(r=－0.701，－0.901，－0.878；Plt;0.05，lt;0.01，lt;0.01)．There was a positive relationship between levels ofGSH in serum and FEV1％pred(r=0．854，Plt;0．01)．In induced sputum，activities of 3’-GCS in all patients was correlated positively with their asthmatic symptom scores and level of MDA f r=0．804，0.926；Plt;0.05，lt;0.叭)．Conclusion γ-GCS and GSH may participate the reaction of
ObjectiveTo determine whether there is an imbalance of KLF2/RelA in peripheral blood neutrophils in patients with bronchial asthma, and explore the relationship between KLF2/RelA imbalance and neutrophil apoptosis.MethodsFrom April 2011 to April 2012, a total of 39 patients with acute attack of asthma in Hunan People's Hospital and Third People's Hospital of Changsha were enrolled, with 13 cases in mild asthma group, 17 cases in moderate asthma group, and 9 cases in severe asthma group. Fifteen healthy subjects were recruited as control group. Peripheral blood were collected from all subjects followed by separation of neutrophils. The apoptosis of neutrophils were measured by flow cytometry. The expression of KLF2 and RelA were detected by Western blot. The relationship between the ratio of KLF2/RelA and neutrophil apoptosis rate was analyzed by Pearson correlation test.ResultsNeutrophil apoptosis rates in the mild, moderate and severe asthma groups [(4.45±0.76)%, (2.10±0.25)%, (1.81±0.67)%, repectively] were lower than that in the healthy control group [(5.36±0.57)%, all P<0.01]. The apoptosis rates of neutrophils in the moderate and severe asthma groups were lower than that in the mild asthma group (bothP<0.01), but there was no significant difference between the moderate asthma group and the severe asthma group (P>0.05). The ratios of neutrophil KLF2/RelA in the mild, moderate and severe asthma groups were lower than that in the normal control group (0.667±0.351, 0.384±0.203, 0.536±0.293vs. 4.038±2.011, all P<0.01). There was no significant difference between the groups of mild, moderate and severe asthma (P>0.05). The neutrophil apoptosis rate was positively correlated with the percentage of neutrophil KLF2/RelA (r=0.592 0, P<0.000 1).ConclusionThere is an imbalance of KLF2/RelA in peripheral blood neutrophils in patients with bronchial asthma, and the imbalance of KLF2/RelA may be the mechanism of apoptosis of peripheral blood neutrophils.
Objective To explore the regulation of peroxisome proliferator-activated receptor γ coactivator 1α( PGC-1α) and NF-E2-related factor 2( Nrf2) on expression of γ-glutamylcysteine synthetase ( γ-GCS) , and their roles in chronic obstructive pulmonary disease( COPD) . Methods Twenty-four SD rats were randomly divided into a COPD group and a normal control group. COPD model was established by intratracheal instillation of lipopolysaccharide ( LPS) and daily exposure to cigarette smog in the COPD group. The lung function was measured and the pathological changes were observed. The protein and mRNA expressions of PGC-1α, Nrf2, and γ-GCS in lung tissue were measured by immunohistochemistry, Western blot, in site hybridization ( ISH) , and reverse transcription-polymerase chain reaction ( RT-PCR ) ,respectively. Results In the COPD group, the pulmonary function ( FEV0. 3, FEV0. 3 /FVC, PEF) damage and lung pathological changes were conformed as morphological characteristics of COPD. The mRNA of PGC-1α and Nrf2 expressed in lung tissues of two group rats in the region consistent with γ-GCS mRNA. The protein and mRNA expressions of PGC-1αand γ-GCS were markedly increased in the COPD group( all P lt;0. 05) ,and the protein expression of Nrf2 was obviously up-regulated ( P lt; 0. 01) , while Nrf2 mRNA had no significant difference between the two groups( P gt;0. 05 ) . Linear correlation analysis showed that the level ofPGC-1αprotein was positively correlated with the levels of Nrf2 protein and mRNA ( r = 0. 775, 0. 515, all P lt; 0. 01) , and the levels of PGC-1αand Nrf2 protein were positively correlated with the levels of γ-GCS protein ( r = 0. 531, 0. 575, all P lt; 0. 01) and mRNA ( r = 0. 616, 0. 634, all P lt; 0. 01) . Conclusions PGC-1α, which may serve as a co-activator of Nrf2, can up-regulate the expression of γ-GCS gene cooperatively with Nrf2 through a common pathway, which might involve in the oxidative and antioxidative mechanism in the pathogenesis of COPD.
Objective To investigate the dynamic expression of small ubiquitin-related modifiers-1 ( SUMO-1) in lung tissue in different phases of rat model of hypoxic pulmonary hypertension( HPH) .Methods Forty Wistar rats were randomly divided into 5 groups, and exposed to normoxia or to normobaric intermittent hypoxia for 3, 7, 14 or 21 days, respectively. Mean pulmonary arterial pressure( mPAP) , right ventricle hypertrophy index ( RVHI) , and the ratio of the vessel wall area to the total area( WA% ) weremeasured. RT-PCR and in situ hybridization were used to determine the mRNA expression of SUMO-1.Immunohistochemistry and Western blot were used to determine the protein expression of SUMO-1. Results The hypoxic rats developed pulmonary vascular remodeling in pulmonary arterioles after 7 days of hypoxia,with WA% and mPAP significantly higher than those in the normal control. Pulmonary vascular remodeling aggravated with much higherWA% and mPAP afer 14 days of hypoxia, and reached the peak afer 21 days of hypoxia. SUMO-1 mRNA and protein expression markedly increased after 3 days of hypoxia, and reached peak after 14 days. After 21 days of hypoxia, SUMO-1 mRNA expression weakened but still higher than that in the normal control ( P lt; 0. 05) , and SUMO-1 protein expression remained stable. SUMO-1 mRNA and protein expression were positively correlated with mPAP, WA% and RVHI( all P lt; 0. 01) . Conclusion SUMO-1 is transcriptionally induced in lung tissue under chronic hypoxia, and thus involves in the pathogenesis of HPH.
Abstract： Objective To investigate the extracellular matrix （ECM） gene expression profile of saphenous vein （SV） in end-stage renal disease （ESRD） patients undergoing coronary artery bypass grafting （CABG）. Methods Sixty-eight patients who were diagnosed as coronary artery disease by coronary angiography and admitted to Department of Cardiovascular Surgery，Zhongshan Hospital of Fudan University from July 2004 to December 2010 were enrolled in this study. According to whether or not they had preoperative ESRD history，all the 68 patients were divided into 2 groups，the ESRD group with 30 ESRD patients who needed maintenance hemodialysis，and the control group with 38 patients without preoperative renal disease. Preoperative clinical data of all the patients were collected in detail. SV samples were obtained at the time of CABG. Microarray，immunohistochemistry and Western blotting were used to investigate the expression profile of ECM genes of SV in ESRD patients undergoing CABG. Results There was no statistical difference in preoperative clinical variables between the 2 groups except the variables which were directly related to their kidney disease （P＞0.05）. There were 16 genes that were up-regulated at least 3-fold and 3 genes that were down-regulated at least 3-fold in the ECM gene expression profile of SV in the ESRD group patients before CABG. The expressions of matrix metalloproteinases-2 （MMP-2） and matrix metalloproteinases-9 （MMP-9） of the ESRD group were significantly higher than those of the control group （2.60±0.50 vs. 0.70±0.16，1.80±0.40 vs. 0.60±0.15，P＜0.01）. The expressions of tissue inhibitor of metalloproteinase-2 （TIMP-2） and tissue inhibitor of metalloproteinase-3 （TIMP-3） of the ESRD group were significantly lower than those of the control group （0.60±0.19 vs. 2.20±0.30，0.90±0.28 vs. 2.40±0.70，P＜ 0.05）. Conclusion A variety of ESRD-related risk factors of cardiovascular diseases may severely influence on the balance of ECM gene expression of SV before CABG，and the resulting imbalance is a risk factor to aggravate SV graft disease after CABG.