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As one of the main members of the Rho GDI dissociation inhibitory factors,D4-GDI inhibits the dissociation of Rho protein and GDP,which is also involved in a wide range of celluar functions,such as cell contraction,adhesion,migration,proliferation and apoptosis.Recently,accumulating evidence has been suggested that D4-GDI is involved in the pathogenesis of several pulmonary diseases,such as lung cancer.Intervention of D4-GDI expression may improve the pathological changes and prognosis of these diseases.
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PRDI-BFI and RIZ homology domain containing proteins (PRDM) play a key role in cell differentiation and proliferation.Most members of the PRDM gene family are tumor suppressor genes which involved in tumorigenesis and abnormal expression in a variety of tumors.Aberrant DNA methylation often silences these genes,which may occur in the early stage of tumor,Cancer can be reversed by demethylation,which provides a new way for cancer treatment.
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Objective To investigate the effect of liver depression (Liver Qi Stagnation) on Th17,Treg,IL-17,IL-10 and airway inflammation in asthmatic rats,and to clarify the immune mechanism of asthma with liver depression.Methods Established the combined with disease and syndrome model of asthma with liver depression.Collected the bronchoalveolar lavage fluid (BALF) to count the total and differential cell.Lung tissue was observed in microscope; the proportion of Th17 cells and Treg cells of CD4 +T cells in peripheral blood was measured by flow cytometry; the levels of IL-17 and IL-10 were determined by ELISA.Results The total number of inflammatory cells[(96.86±4.43)× 107/L,(88.22±3.22)× 107/L],the proportion of eosinophils [(27.58 ±4.65) ％,(22.67±2.43) ％],Th17 cells[(6.86±0.98) ％,(6.01 ±0.77) ％] and IL-17 level [(48.88± 8.06)pg/ml,(43.24± 6.32) pg/ml] of asthma in liver depression group and asthma group were significantly higher than the control group [(30.58 ± 2.49) × 107/L,(0.78 ± 0.12) ％,(2.80± 0.82) ％,(24.11 ±3.40)pg/ml]; Treg cells [(3.09±0.55) ％,(3.96±0.66) ％] and IL-10 level [(19.79±2.80) pg/ml,(20.29±3.12) pg/ml] were significantly lower than the control group [(8.02± 1.26) ％,(30.79 ± 4.01) pg/ml].The total number of inflammatory cells (96.86 ±4.43) × 107/L,the proportion of eosinophils (27.58±4.65) ％ and Th17 cells(6.86±0.98) ％ and IL-17 level (48.88±8.06)pg/mL of asthma in liver depression group were significantly higher than the asthma group (88.22 ± 3.22) × 107/L,(22.67 ± 2.43) ％,(6.01 ± 0.77) ％,(43.24 ± 6.32) pg/ml;the proportion of Treg cells (3.09 ±0.55)％ was significantly lower than the asthma group (3.96± 0.66)％; and the lung histopathology symptoms was more severe than asthma group.Conclusion Liver Qi Stagnation can promote the inflammation of asthma,the imbalance of Th17/Treg and IL-17 level to aggravate the asthma.Liver depression is one of the major internal factors in recurrent episodes of asthma.
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AIM: To explore the effects of peroxisome proliferator-activated receptor γ (PPARγ) on the activity and expression of γ-glutamylcysteine synthetase (γ-GCS), and its role in rats with chronic obstructive pulmonary disease. METHODS: COPD model was established by the method of combining fumigation and lipopolysaccharide (LPS) intra-tracheal dripping. Meanwhile, some of the COPD rats were administered with rosiglitazone (RGZ), a PPARγ activator. The pulmonary function and the pathological changes were determined. ROS content and γ-GCS activity in lung tissues were detected. The levels of PPARγ, γ-GCS mRNA and protein expression in lung tissues were measured by immunohistochemistry, Western blotting, in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The pulmonary function (FEV_(0.3), FEV_(0.3)/FVC%, PEF) were significantly improved in RGZ group compared to COPD group. Under light microscope, lung pathological changes in COPD group conformed to pathological features of COPD. The pathological changes of lung tissue were obviously reduced in RGZ group compared to COPD group. In RGZ group, ROS content was obviously reduced and γ-GCS activity significantly increased compared to COPD group. Protein and mRNA expressions of PPARγ and γ-GCS in COPD group significantly higher than those in control group (all P<0.01), and those in RGZ group was markedly increased compared to COPD group (all P<0.05). Linear correlation analysis showed that PPARγ protein was positively correlated with γ-GCS activity (r=0.634, P<0.01), and was no significantly correlated with ROS content (r=0.214, P>0.05). PPARγ protein was positively correlated with γ-GCS protein and mRNA (r=0.553, r=0.442, all P<0.01). CONCLUSION: PPARγ activation by RGZ reduces the extent of COPD oxidant/antioxidant imbalance, which plays an important role in the prevention and treatment of COPD. In addition, PPARγ may play an important antioxidant protection role by reducing ROS production, and increasing activity and gene expression of γ-GCS in the lung.
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AIM: To investigate the expression of PPARγ and Nrf2/γ-GCS-h in inflammatory cells in bronchoalveolar lavage fluid(BALF) of guinea pig with bronchial asthma of acute episode, and to explore the roles of PPARγ on Nrf2/γ-GCS-h expression. METHODS: Forty adult male guinea pigs were randomly divided into 4 groups (10 guinea pigs in each group): control group (group A), asthmatic group (group B), dexamethasone treatment group (group C) and rogridone treatment group (group D). The asthmatic model was established by an ovalbumin challenge method. BALF was collected, and the total cell count and the proportion of the inflammatory cells were measured. After centrifugation, the concentrations of ROS and MDA in the clear supernatant were detected. The methods of in situ hybridization and immunohistochemistry were used for detecting the expression of PPARγ and Nrf2/γ-GCS-h at mRNA and protein levels. RESULTS: The proportion of eosinophils (EOS) in BALF in group B was significantly higher than that in groups A, C and D (P<0.01). The concentrations of ROS and MDA in BALF of group B was the highest. The difference of ROS and MDA was statistically significant (all P<0.05) as compared to the control. The results of immunohistochemistry and in situ hybridization indicated that the A value was the lowest in group B as compared to that in groups A, C and D (all P<0.01). In group B, the positive correlations were observed between PPARγ and Nrf2/γ-GCS-h, between γ-GCS-h and Nrf2. A negative correlation between the proportion of EOS in BALF and the expression of PPARγ and Nrf2/γ-GCS-h was also observed (all P<0.05). CONCLUSION: In acute asthmatic models induced by ovalbumin, the expression of PPARγ and Nrf2/γ-GCS-h is decreased, and PPARγ may up-regulate the expression of Nrf2/γ-GCS-h to inhibit the inflammatory and oxidative reactions, indicating a new way for prevention and treatment of bronchial asthma.
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Objective To observe the effect of PDK ( phosphoinositol -3-kinase, PI3K)/Akt-Nrf2 (Nuclear factor -E2 related factor) signal pathway on γ-glutamylcysteine synthetase (γ-GCS) in the bronchial epithelial cells of rats treated with cigarette smoke extract (CSE). Methods The bronchial epithelial cells were dealt with 10% concentration of CSE for different time and pretreated with PI3k inhibitor (LY294002). The expressions of Nrf2,p-Akt and γ-GCS proteins were examined by immunocytochemistry, flow cytometry, immunofluorescence and western blot. The expressions of γ-GCS mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Reduced glutathione(GSH) content and the lev-el of γ-CCS activi-ty were examined. Results GSH content in CSElh group was significantly decreased, but still higher in CSE3 and 6 groups compared to the control. Nrf2 protein mainly located in the cytoplasm. Nrf2 plasmosin mainly increased in the nucleus in control group, and Nrf2 nucleic protein significantly enhanced in CSE1, 3 and 6 groups. P-Akt protein was up-regulated at lh, reached its peak at 3h, declined slightly at 6h after exposure to CSE. The tendency of the percentage of p-Akt positive cells was as same as p-Akt protein. γ-GCS mRNA, protein and activity, gradually increased in CSE lh, CSE 3h,CSE 6h groups. Pretreated with LY294002, the expression of p-Akt protein was markedly decreases, while Nrf2 plasmosin expressed strongly, and γ-GCS mRNA, protein, activity and GSH content were significantly decreased compared to CSE3h group. Linear correlation analysis demonstrated that there were a positive correlation among Nrf2 and γ-GCS,γ-GCS activity, and among p-Akt and Nrf2,GSH,γ-GCS,γ-GCS activity. Conclusion P13K/Akt signal path might participate in Nrf2 nuclear translocation via regulating the expression of γ-GCS.
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Objective To investigate the relationship among the concentration of plasma interleukin (IL)-10 and plasma IL-13 and the clinical therapeutic effect in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods Thirty-six AECOPD inpatients were enrolled in this study.Blood samples of the subjects were collected as soon as hospitalization, and plasma IL-10 and IL-13concentrations were analyzed by enzyme-linked immunosorbent assay.The clinical manifestations of subjects were quantified by a special designed score standard, and were evaluated at the time points of hospitalization, 48 hours treatment and 96 hours treatment.The therapeutic effect was evaluated by clinical manifestations score combined with pulmonary ventilation functional parameter.Eventually, the correlation among the concentration of IL-10 concentration and IL-13 and the clinical therapeutic effect were analyzed.Results The correlation coefficients between clinical manifestations score decrease and pulmonary ventilation function improvement with plasma IL-10 concentration after 48 hours treatment were 0.85 and 0.48 respectively,then, 0.64 and 0.52 after 96 hours treatment.The correlation coefficients between clinical manifestations score decrease and pulmonary ventilation function improvement with plasma IL-13 concentration after 48hours treatment were -0.41 and -0.34, after 96 hours treatment , correlation coefficients between clinical manifestations score decrease with plasma IL-13 concentration was -0.36.All of the above correlation coefficients were statistically significant.Conclusion Plasma IL-10 concentration was positively, whereas, IL-13 concentration was negatively correlated with the therapeutic effect in AECOPD patients.
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Objective To survey the prevalence of chronic obstructive pulmonary disease (COPD)in urban areas of Hunan province and relevant risk factors and provide a basis of the prevention and treatment for COPD. Methods A questionnaire survey was conducted among 4248 residents, aged over 15, by a simple cluster random sampling method in Changsha, Hunan, Wulipai street North Station community. All the respondents filled out an unified epidemiological survey questionnaire. All of the respondents received examination for lung function. Those respondents showed FEV1/FVC ＜70% were further examined by ECG,X ray inspection for differential diagnosis. The data of epidemiological survey was analyzed by multivariate logistic regression method. Results The response rate was 92%. The total prevalence of COPD was 4. 81%.The prevalence of COPD in the males was 6. 6%, and 3. 0% in the females. The prevalence of COPD in the males was significantly higher than that in the females (x2 = 29. 915, P ＜ 0. 01). The prevalence increased with age increasing (P ＜0. 01). The more the education was, the lower the prevalence of COPD was. Risk factors analyzed with non-conditional logistic were as follow. The odd ratio (OR) for COPD in the age was 1.92(P ＜0. 01) and the odd ratio (OR) for COPD in the sex was 1.81 (P ＜0. 01). The weak lighting in house increased the risk with the OR of 4. 25(P ＜0. 01) and pet feeding further increased the risk with the OR of 12.08(P ＜0. 01). The odd ratio (OR) for COPD in the smokers was 1.74(P ＜0. 01) and the prevalence of COPD was related with smoking intensity (branch years of cigarette). Smoking intensity above 500 increased the risk of COPD. The passive smoking increased the risk with the OR of 16. 39(P ＜0. 01). The odd ratio (OR) for COPD in the paternal family history with chronic pulmonary disease was 2. 13(P ＜0. 01) and 2. 11 (P ＜ 0. 01) in the maternal family history. The odd ratio (OR)for COPD in the education degree was 0. 52(P ＜ 0. 01). Conclusions The prevalence of COPD was high in Changsha city, which might be attributed to the risk factors such as house lighting, pet feeding, cooking,aged, male, smoking, passive smoking, and family history. The education degree was the protective factor of COPD. We should intervene the relevant risk factors of COPD so that the prevalence of COPD might be cut down.
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Whether surfactant protein B (SP-B)-18A/C and 1580C/T polymorphism were associated with susceptibility to chronic obstructive pulmonary disease (COPD) in Chinese Han population was investigated. After genomic DNA was isolated from blood of COPD smokers and control smokers, the genotypes of SP-B-18A/C and SP-B1580C/T polymorphism loci were determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) respectively. The results showed that there was significant difference in genotypes distribution frequency of SP-B1580C/T polymorphism locus between COPD smokers and control smokers. C-->T mutation rate (including TT homozygote and CT heterozygote) in COPD smokers was higher than in control smokers (57.9% vs 41.7%, chi2 = 4.93, P<0.05), whereas there was no significant difference in genotypes distribution frequency of SP-B1580-18A/C locus between COPD smokers and control smokers. The allele frequency (29.1%) of SP-B1580-18A/C locus is lower than T allele (70.9%) in Chinese Han Population, and the distribution was different from that in Mexican, in which, the A and T allele frequencies were 85% and 15% respectively. It was concluded that SP-B1580 T allele was probably associated with increased susceptibility to COPD in Chinese Han population; The polymorphism of SP-B-18A/C locus maybe varied with race.
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Alleles , China/ethnology , Genetic Predisposition to Disease/genetics , Genotype , Polymorphism, Genetic/genetics , Pulmonary Disease, Chronic Obstructive/genetics , Pulmonary Surfactant-Associated Protein B/genetics , Pulmonary Surfactant-Associated Protein B/physiology , Smoking/geneticsABSTRACT
Whether surfactant protein B (SP-B)-18A/C and 1580C/T polymorphism were associated with susceptibility to chronic obstructive pulmonary disease (COPD) in Chinese Han population was investigated. After genomic DNA was isolated from blood of COPD smokers and control smokers, the genotypes of SP-B-18A/C and SP-B1580C/T polymorphism loci were determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) respectively. The results showed that there was significant difference in genotypes distribution frequency of SP-B1580C/T polymorphism locus between COPD smokers and control smokers. C-->T mutation rate (including TT homozygote and CT heterozygote) in COPD smokers was higher than in control smokers (57.9% vs 41.7%, chi2 = 4.93, P<0.05), whereas there was no significant difference in genotypes distribution frequency of SP-B1580-18A/C locus between COPD smokers and control smokers. The allele frequency (29.1%) of SP-B1580-18A/C locus is lower than T allele (70.9%) in Chinese Han Population, and the distribution was different from that in Mexican, in which, the A and T allele frequencies were 85% and 15% respectively. It was concluded that SP-B1580 T allele was probably associated with increased susceptibility to COPD in Chinese Han population; The polymorphism of SP-B-18A/C locus maybe varied with race.
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Female , Humans , Male , Alleles , China , Ethnology , Genetic Predisposition to Disease , Genetics , Genotype , Polymorphism, Genetic , Genetics , Pulmonary Disease, Chronic Obstructive , Genetics , Pulmonary Surfactant-Associated Protein B , Genetics , Physiology , Smoking , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of hypoxia-inducible factor 1 alpha (HIF-1alpha) and inducible nitric oxide synthase (iNOS) genes in rats' pulmonary arteries in different phases of hypoxia-induced pulmonary hypertension development.</p><p><b>METHODS</b>Models of chronic hypoxic pulmonary hypertension rat were duplicated by intermittent hypoxia. Mean pulmonary arterial pressure (mPAP) was measured by right-heart catheterization. HIF-1alpha and iNOS messenger ribonucleic acid (mRNA) were detected by in situ hybridization. HIF-1alpha and iNOS protein were measured by immunohistochemical analysis.</p><p><b>RESULTS</b>Expression of HIF-1alpha protein was upregulated in pulmonary arterial tunica intimae of all hypoxic rats. In pulmonary arterial tunica media, the level of HIF-1alpha protein was markedly upregulated at days 3 and 7 of hypoxia (P < 0.01), then tended to restore at 14 days and 21 days. HIF-1alpha mRNA levels in pulmonary arteries of rats began to increase significantly at day 14 of hypoxia (P < 0.01). Expression of iNOS mRNA and protein in pulmonary arteries of rats were upregulated by hypoxia for 3 days (P < 0.01), then reached its peak and maitained the same level while the extension of hypoxia. Linear correlation analysis showed that iNOS protein was associated with both mean pulmonary arterial pressure (r = 0.74, P < 0.01) and hypoxic pulmonary vascular remodeling (r = 0.78, P < 0.01), whereas the inverse was associated with HIF-1alpha protein (r = -0.52, P < 0.01).</p><p><b>CONCLUSIONS</b>HIF-1alpha and iNOS are both involved in the pathogenesis of hypoxia-induced pulmonary hypertension in rat. HIF-1alpha protein may upregulate the expression of iNOS gene by transcriptional activation; in addition, iNOS protein may inhibit the expression of HIF-1alpha protein.</p>
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Animals , Male , Rats , Gene Expression Regulation, Enzymologic , Hypertension, Pulmonary , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit , Nitric Oxide Synthase , Genetics , Nitric Oxide Synthase Type II , Pulmonary Artery , RNA, Messenger , Rats, Wistar , Transcription Factors , Genetics , Physiology , Up-RegulationABSTRACT
AIM: To investigate atypical protein kinase C(aPKC), extracellular signal regulated kinnase (ERK) regulating NF-E2-related factor 2 (NRF2)-?-gutamylcysteine synthetase (?-GCS) and the effect on lung of rats with chronic obstructive pulmonary disease (COPD). METHODS: The rat COPD model was established by intratracheal instillation of lipopolysaccharide twice and exposure to cigarette smoke daily. The ?-GCS activity was measured. The expression of ?-GCS mRNA in lung tissue was examined by in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR). The protein expressions of p-aPKC?/?, p-ERK, NRF2 and ?-GCS in lung tissue were detected by immunohistochemistry (IH) and Western blotting, respectively. RESULTS: (1) The ?-GCS activity was higher in COPD group than that in control group. (2) The expression of ?-GCS mRNA in the COPD group was stronger than that in control group. ISH showed that the ?-GCS mRNA was expressed in alveolar epithelium and bronchiolar smooth muscle cell in the COPD group. (3) The protein expressions of p-aPKC, p-ERK, NRF2, ?-GCS were significantly higher than those in control group. IH showed that p-aPKC, p-ERK, NRF2, ?-GCS proteins were expressed in alveolar and bronchiolar epithelium in the COPD group. (4) There was a positive correlation between NRF2 and ?-GCS. ?-GCS mRNA, p-aPKC?/?, p-ERK were also positively correlated with NRF2. CONCLUSION: By upregulating the signal transduction of NRF2-?-GCS, the ERK and aPKC?/? may play an important role in the mechanism of COPD formation.
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AIM: To investigate Nrf2 that regulates the expression of ?-glutamylcysteine synthetase(?-GCS) in the inflammatory cells in bronchial asthma guinea pig bronchoalveolar lavage fluid(BALF).METHODS: Adult male guinea pigs were randomly divided into control group(group A),asthmatic group(group B) and dexamethasone group(group C).Asthmatic model was established by the method of ovalbumin challenge.MDA concentration in the lung tissue homogenate was detected.The total cell count and the proportion of inflammatory cells in BALF were measured.The methods of immunohistochemistry and in situ hybridization were used for detection of the protein and mRNA expressions of Nrf2,Bach1 and ?-GCS.RESULTS:(1) The proportion of eosinophils(EOS) in BALF and the MDA concentration of the lung tissue in group B were higher than those in group A and group C.(2) The result of in situ hybridization indicated that the A value of ?-GCS was the highest in group A compared to group B and group C,but the A value of Nrf2 and Bach1 in 3 groups has no statistical significance.(3) Immunohistochemistry indicated that the A value of ?-GCS in group B was lower than that in group A.The positive rate in cell nucleus of Nrf2 in group B was lower than that in group A and group C.The positive rate in cell nucleus of Bach1 in group B was higher than that in group A and group C.(4) The mRNA expression of ?-GCS(A value) showed positive correlation with the positive rate in cell nucleus of Nrf2 and negative correlation with the positive rate in cell nucleus.A negative correlation between the proportion of EOS in BALF of group B and the ?-GCS mRNA was observed.CONCLUSION: There is disequilibrium between oxidation and anti-oxidation in bronchial asthma guinea pig.Inflammatory reaction decreases the expression of ?-GCS in the inflammatory cells in bronchial asthma guinea pig.Dexamethasone regulates the nuclear translocation of Nrf2/Bach1 and increases the expression of ?-GCS.