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Multidrug resistance associated protein-1 (MRP1) and Notch signaling are closely related and both play a critical role in chronic obstructive pulmonary disease (COPD) establishment and progression. The aim of our work was to test whether Notch1 is involved in allyl isothiocyanate (AITC) induced MRP1 expression. We used cigarette smoke extract (CSE) to simulate the smoking microenvironment in vitro. The results demonstrated that CSE led to apoptosis as well as reduced the expression of Notch1, Hes1, and MRP1, while AITC significantly reversed this downregulation. Transfected with Notch1 siRNA downregulated MRP1 expression and activity, aggravated the suppression effect by CSE, and abolished the AITC-induced Notch1, Hes1, and MRP1 expression. Validation of the correlation between Notch1 and MRP1 was implemented by gel-shift assays (electrophoretic mobility shift assay). The result revealed an interaction between a specific promoter region of MRP1 and the intracellular domain of Notch1. In conclusion, Notch1 signaling positively regulated MRP1 in 16HBE cells and AITC induced MRP1 expression and function may be attributed to Notch1 signaling. These findings show that Notch1 and MRP1 might have a potential protective effect in the COPD process and become a new therapeutic target for COPD or other lung diseases. It also provides a theoretical basis for the therapeutic effects of AITC. Key words: allyl isothiocyanate (AITC), chronic obstructive pulmonary disease (COPD), cigarette smoke extract (CSE), Notch1 signaling, multidrug resistance associated protein-1(MRP1). La proteine 1 associee a la multiresistance aux medicaments (ou MRP1 pour <>) et la voie de signalisation Notch sont intimement liees et jouent toutes deux un role essentiel dans la presentation et l'evolution de la maladie pulmonaire obstructive chronique (MPOC). Nos travaux avaient pour but d'evaluer si la proteine Notch1 joue un role dans l'expression de la MRP1 engendree par l'isothiocyanate d'allyle (ITCA). Nous avons simule un microenvironnement de tabagisme in vitro a l'aide d'extraits de fumee de cigarette (EFC). Les resultats ont montre que les EFC menaient a l'apoptose, ainsi qu'a une diminution de l'expression des proteines Notch1, Hes1 et MRP1, tandis que l'ITCA inversait cette regulation a la baisse de facon notable. La transfection avec du siRNA du gene Notch1 entrainait une regulation a la baisse de l'expression et de l'activite de la proteine MRP1, une aggravation de l'effet suppresseur des EFC, avec une inhibition complete de l'expression des proteines Notch1, Hes1 et MRP1 produites par l'ITCA. Nous avons valide la correlation entre les proteines Notch1 et MRP1 a l'aide de la technique de retard sur gel (ou EMSA pour <>). Les resultats ont revele la presence d'une interaction entre une region promotrice specifique du gene MRP1 et le domaine intracellulaire de la proteine Notch1. En conclusion, la signalisation par la proteine Notch1 entrainait une regulation avantageuse du gene MRP1 dans les cellules 16HBE, et l'expression ainsi que le fonctionnement de la proteine MRP1 produite par l'ITCA pourraient etre attribues a la signalisation par la proteine Notch1. Ces resultats montrent que les proteines Notch1 et MRP1 pourraient avoir un effet protecteur eventuel contre les processus de la MPOC et devenir une nouvelle cible therapeutique dans la MPOC ou d'autres maladies pulmonaires. Ils fournissent aussi une base theorique pour expliquer les effets therapeutiques de l'ITCA. [Traduit par la Redaction] Mots-cles: isothiocyanate d'allyle (ITCA), maladie pulmonaire obstructive chronique (MPOC), extraits de fumee de cigarette (EFC), signalisation Notch1, proteine 1 associee a la multiresistance aux medicaments (MRP1)., Introduction Chronic obstructive pulmonary disease (COPD) is one common and serious disease characterized by irreversible and progressive airflow obstruction, which is associated with abnormal chronic inflammatory responses in the airway [...]

Chronic obstructive pulmonary disease (COPD) exacerbations accelerate loss of lung function and increased mortality. The complex nature of COPD presents challenges in accurately predicting and understanding frequent exacerbations. The present study aimed to assess the metabolic characteristics of the frequent exacerbation of COPD (COPD-FE) phenotype, identify potential metabolic biomarkers associated with COPD-FE risk and evaluate the underlying pathogenic mechanisms. An internal cohort of 30 stable patients with COPD was recruited. A widely targeted metabolomics approach was used to detect and compare serum metabolite expression profiles between patients with COPD-FE and patients with non-frequent exacerbation of COPD (COPD-NE). Bioinformatics analysis was used for pathway enrichment analysis of the identified metabolites. Spearman's correlation analysis assessed the associations between metabolites and clinical indicators, while receiver operating characteristic (ROC) analysis evaluated the ability of metabolites to distinguish between two groups. An external cohort of 20 patients with COPD validated findings from the internal cohort. Out of the 484 detected metabolites, 25 exhibited significant differences between COPD-FE and COPD-NE. Metabolomic analysis revealed differences in lipid, energy, amino acid and immunity pathways. Spearman's correlation analysis demonstrated associations between metabolites and clinical indicators of acute exacerbation risk. ROC analysis demonstrated that the area under the curve (AUC) values for D-fructose 1,6-bisphosphate (AUC=0.871), arginine (AUC=0.836), L-2-hydroxyglutarate (L-2HG; AUC=0.849), diacylglycerol (DG) (16:0/20:5) (AUC=0.827), DG (16:0/20:4) (AUC=0.818) and carnitine-C18:2 (AUC=0.804) were >0.8, highlighting their discriminative capacity between the two groups. External validation results demonstrated that DG (16:0/20:5), DG (16:0/20:4), carnitine-C18:2 and L-2HG were significantly different between patients with COPD-FE and those with COPD-NE. In conclusion, the present study offers insights into early identification, mechanistic understanding and personalized management of the COPD-FE phenotype. [ABSTRACT FROM AUTHOR]

The differences of transitional components and metabolic processes of Huatan Jiangqi Capsules(HTJQ) in rats under normal physiological and pathological conditions of COPD were analyzed by UPLC-Q-TOF-MS. The rat COPD model was established by passive smoking and intratracheal instillation of lipopolysaccharide. After the normal and COPD model rats were douched with HTJQ, the blood was collected from hepatic portal vein and the drug-containing serum samples were prepared by methanol precipitation of protein. Then, 10 batches of drug-containing serum samples of HTJQ were prepared and analyzed by UPLC serum fingerprint to evaluate the quality and stability of drug-containing serum samples. UPLC-Q-TOF-MS was used to collect the mass spectrometric information of the transitional components. Twenty-eight transitional components of HTJQ in normal rats and 25 transitional components of HTJQ in COPD model rats were identified by UPLC-Q-TOF-MS. Under pathological and physiological conditions, there were not only the same transitional components in rat serum, but also corresponding differences. Further studies showed that there were also differences in the metabolic process of transitional components between the two conditions. In normal rats, most of the metabolic types of transitional components were phase I reactions. In COPD model rats, phase Ⅰ reactions decreased and phase Ⅱ reactions increased correspondingly. With UPLC-Q-TOF-MS technology, the differences of transitional components and the metabolism process of HTJQ in rats under normal physiological and pathological conditions were analyzed. The results showed that types of transitional components and the activity of some metabolic enzymes would be changed in COPD pathological state, which would affect the metabolic process of bioactive components in vivo. It laid a foundation for further elucidating the metabolic process and pharmacodynamic substance basis of HTJQ.

Background: Chronic obstructive pulmonary disease (COPD) is predicted to become the fifth leading cause of disability and the third leading cause of death around the world by 2020. Though it is potentially treatable and preventable, evidence of brain structural alterations in COPD remains sparse and conflicting. We aim to investigate the effect of Liuweibuqi capsules on CD4 + CD25 + Forkhead box protein 3 + (Foxp3 + ) regulatory T cells (Tregs), helper T cells (Th) and lung function in patients with stable COPD complicated with lung Qi deficiency., Methods: COPD patients with lung Qi deficiency [458] were assigned into non-smoking COPD (NS-COPD), non-smoking control (NS-control), smoking COPD (S-COPD) and smoking control (S-control) groups, and healthy volunteers [245] into the non-smoking healthy (NSH) and smoking healthy (SH) groups. Levels of inflammatory cytokines were detected by Enzyme-linked immunoassay (ELISA). Contents of inflammatory cells, inflammatory marker, and CD4 + CD25 + Fox3 + Tregs were measured by flow cytometry. FEV1/FVC (%) and FEV1 (%) were detected by pulmonary function test apparatus. Correlation between FEV1 (%) and Th1, Th2, Th17, Th1/Th2 or CD4 + CD25 + Fox3 + Tregs was analyzed by Spearman rank correlation test. The related factors affecting treatment efficacy was assessed by logistic analysis., Results: COPD patients and smoking people showed higher level of INF-γ, IL-4, IL-17, Th1, Th2, Th17 and Th1/Th2 but lower level of CD4 + CD25 + Fox3 + Tregs. Liuweibuqi capsules could decrease level of inflammatory cells, cytokines, and markers (especially Th17 and IL-17), and increase level of CD4 + CD25 + Fox3 + Tregs. FEV1 (%) negatively correlated with Th1, Th2, Th17 and Th1/Th2 but positively correlated with CD4 + CD25 + Fox3 + Tregs, and smoking may strengthen their correlation, but Liuweibuqi capsules may weaker their correlation. Levels of inflammatory cytokines, cells, marker, CD4 + CD25 + Fox3 + Tregs, FEV1/FVC (%), FEV1 (%), smoking and Liuweibuqi capsules are factors affecting efficacy., Conclusions: Taken together, our data support the notion that smoking is an important factor to induce and aggravate COPD. Liuweibuqi capsules could stimulate proliferation of CD4 + CD25 + Fox3 + Tregs and decrease Th17 expression to improve the lung function in stable COPD patients with lung Qi deficiency, and it had obvious efficacy for smoking COPD patients., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest to declare.

To investigate the ATP-sensitive potassium channel (KATP channel) protein expressions during different periods under hypoxia condition and explore the effect of Qibai Pingfei capsule medicated serum (hereinafter referred to as QBPF) on the correlation between the protein expressions of KATP channel and nitric oxide in rat pulmonary arterial smooth muscle cells(PASMCs). Qibai Pingfei capsules were given to SD rats via continuous gavage for 10 days to obtain QBPF. Primary rats PASMCs were cultured by the direct adherent culture method. Western blot was applied to detect the protein expression levels of KATP channel (Kir6.1 and SUR2B) in PASMCs. Then the noncompetitive inhibitor of NO synthase--Nω-nitro-L-arginine methyl ester(L-NAME) and KATP channel inhibitor--glyburide(GLYB) were applied respectively to evaluate the effect of QBPF on the protein expressions of KATP channel. The protein expressions of Kir6.1 and SUR2B were increased after 6-hour hypoxia treament, peaked at the 24-hour hypoxia treament, and decreased in both 48-hour and 72-hour hypoxia groups. Especially, QBPF could further up-regulate the Kir6.1 and SUR2B protein expressions under 24-hour hypoxia condition; however, such up-regulation effect could be blocked by KATP channel inhibitor GLYB and NO specific inhibitor L-NAME, indicating that QBPF played the role of opening KATP channel. The regulatory mechanism was probably associated with up-regulating KATP channel protein expression via NO relative pathway, involving pulmonary vasodilation, and thus relieving the occurence and development of COPD., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To observe the changes in proportion of CD4 + CD25 + Foxp3 + regulatory T cells (Treg) in peripheral blood of different stages of CPOD and intervention of Qibai Pingfei Capsules., Methods: The rats were randomly divided into three groups,including normal group, COPD group and Qibai Pingfei Capsules(2. 88 g/kg)group. At the end of 7,14,21 and 28 days,eight rats were sacrificed in each group. CD4 + CD25 + Foxp3 + Treg cells in peripheral blood were measured by flow cytometry method., Results: Compared with normal group, the percentages of CD4 + % in peripheral blood were not significantly different at the end of 7, 14, 21 and 28 days. However, CD4 + CD25 + % and CD4 + CD25 + Foxp3 +/CD4 + were significantly increased and CD4 + CD25 + Foxp3 + Treg% were significantly decreased at the end of 14,21 and 28 days. Compared with model group, CD4 + CD25 + %, CD4 + CD25 + Foxp3 +/CD4 + were significantly decreased, CD4 + CD25 + Foxp3 + Treg % were significantly increased at different stages of CODP., Conclusion: Immune disorders may exist in COPD, and Treg cells may be involved in the process of COPD. Meanwhile,the protective effect in COPD rats of Qibai Pingfei Capsules may be associated with improving the percentage of suppressive CD4 + CD25 + Foxp3 + Tregs.

Objective: To determine T box expressed in T cells (T bet), GATA binding protein-3 (GATA 3), and retinoid-related orphan nuclear receptor gammat (RORgammat) in rats with chronic obstructive pulmonary disease (COPD)., Methods: Thirty rats were randomly divided into a control and a COPD group. The COPD model was established through smoking and lipopolysaccharide (LPS) tracheal instillation. Pulmonary function of the rats was measured 28 d after the establishment of the COPD model by a spirometer. Enzyme linked immunosorbent assay was performed to detect serum γ interferon (IFN-γ), interleukin (IL)-4, and IL-17. The expressions of T-bet, GATA-3, and RORgammat protein in lung tissues were determined by Western blot., Results: Compared with the controls, the COPD rats had decreased pulmonary function and expression of serum IL-4, and increased INF-γ, IL- 17, Th1/Th2, T-bet, T bet /GATA-3, and RORgammat protein (P<0. 05). Forced expiratory volume in 0. 3 seconds (FEV 0.3) was negatively correlated with INF γ and T-bet/GATA-3. Forced vital capacity (FVC) was positively correlated with IL 4. FEV0.3/FVC was negatively correlated with Thl/Th2, T-bet and T-bet/GATA-3. Peak expiratory flow (PEF) was negatively correlated with IL-17, T bet, and RORgammat (P<0. 05). Thl/Th2 was positively correlated with T bet/GATA-3. IL-17 was positively correlated with RORgammat. T bet/GATA-3 was positively correlated with RORgammat (P<0. 05)., Conclusion: Imbalanced regulation of T bet / GATA 3 and RORgammat on Th1/Th2 and Th17 cells is associated with the occurrence of COPD.

Objective: By using the metabonomics method, to study the plasma metabonomics of the stable phase chronic obstructive pulmonary disease (COPD) patients of Fei-qi deficiency syndrome (FQDS), and of Chinese materia medica (CMM) intervention, thus exploring possibly existent biomarkers., Methods: Forty stable phase COPD patients of FQDS were recruited as Group A. Liuwei Buqi Capsule (LWBQC) was given to them as intervention. A healthy control group (Group B, 37 cases) was set up. The pulmonary function test was performed on patients in Group B and Group A before and after intervention. The plasma metabolites were detected using high performance liquid chromatography-mass spectrometry (HPLC-MS/MS). Statistical data were analyzed using principal component analysis (PCA) and partial least squares (PLS). The original spectrum and data of plasma metabonomics were compared between the two groups. The whole spectrum amino acid metabonomics tests were performed in the two groups., Results: Compared with Group B, the pulmonary function significantly decreased before intervention in Group A (P < 0.05). The pulmonary function was more mildly improved after 30 days' treatment than before treatment, but still lower than it in Group B (P < 0.05). The metabolic spectrum before treatment in Group A was significantly different from Group B, but showing regressive trend to Group B after treatment. Fifteen possible disease markers were found in COPD patients of FQDS. Results of the whole spectrum of amino acid metabolomics showed different features., Conclusions: The changes of metabolic spectrum and amino acids could be found in the stable phase COPD patients of FQDS using plasma metabonomics, and potential markers could be detected. The intervention of the stable phase COPD patients of FQDS by Chinese medicine could brought positive changes in the metabolic profiling and amino acid metabolism.

Objective: To study T lymphocyte related genes with differential expression in patients with chronic obstructive pulmonary disease (COPD) of Fei-qi deficiency (FQD) syndrome type by gene chips., Methods: Lymphocytes in peripheral blood were isolated by Ficoll technique from blood samples collected from COPD patients of FQD syndrome type, Fei-yin deficiency (FYD) syndrome type, and also from healthy subjects for control. They were sorted and purified by flow cytometry, and the different expressed genes were screened from them by gene chip technique., Results: There were 15 genes with high differential expression between patients of FQD type and those of FYD syndrome type, and between patients of FQD type and healthy subjects., Conclusion: Gene chip technique could be used for studying the gene expression profiles of TCM syndrome, and the T-lymphocyte related genes with differential expression in COPD patients with FQD were screened preliminarily.

Objective: To establish an experimental animal model of Alzheimer's disease (AD) with clinical and pathological specificity conformed to AD in human, and to observe the effect of Zhinao Capsule (ZNC) on learning, memory and patho-morphological parameters in the model., Methods: The experimental AD model of rats was improved and established by combined injection of beta-amyloid protein (beta-AP) to lateral ventricle and transforming growth factor beta 1 (TGF beta 1) to brain. The learning and memory function of the model rats were tested by step-through test and water-maze test, and the number and cross area of beta-AP deposited macula in cerebral cortex and CA1 region of hippocampus were estimated quantitatively using immunohistochemical method and image pattern analysis., Results: The improved AD animal model showed both the specificity of behavior (learning and memory impairments) and the typical pathological specificity (beta-AP deposited macula). ZNC could effectively improve the impairment of learning and memory and reduce the number and cross area of beta-AP deposited macula in cerebral cortex and hippocampus CA1 region in the AD model rats., Conclusion: The AD rat model induced by the combined injection of beta-AP and TGF beta 1 is a good animal model simulated to the clinical reality, which could be used to screen and evaluate the anti-dementia agents. ZNC could display anti-dementia effect of the AD model rats.