1.The regulation of Nrf2/Bachl on gamma-glutamylcysteine synthetase in inflammatory cells from bronchial asthmic patient sputum.
Xiu-feng ZHANG ; Ai-guo DAI ; Rui-cheng HU
Chinese Journal of Applied Physiology 2010;26(2):226-228
Adult
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Asthma
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metabolism
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physiopathology
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Basic-Leucine Zipper Transcription Factors
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genetics
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physiology
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Fanconi Anemia Complementation Group Proteins
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genetics
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physiology
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Female
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Glutamate-Cysteine Ligase
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metabolism
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Humans
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Inflammation
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metabolism
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pathology
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Male
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NF-E2-Related Factor 2
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genetics
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physiology
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RNA, Messenger
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genetics
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metabolism
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Sputum
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cytology
2.Clinical study on correlation between the level of CD4+CD25+regulatory T cells in peripheral blood and APACHE Ⅱ scores in patients with sepsis
Xingui DAI ; Yuhang AI ; Yeping CAI ; Yong GUO ; Jiafen HE
Chinese Journal of Postgraduates of Medicine 2008;31(34):16-18
Objective To evaluate the correlation between the level of CD4+CD25+regulatory T cells in periphend blood and disease severity in patients with sepsis.Methods Thirty-six septic patients and 5healthy controls were enrolled.Septic patients were divided into sepsis group(n=10),severe sepsis group (n=15)and septic shock group(n=11).The lymphocyte was seperated from peripheral blood and marked by PE-CD4 and FTTC-CD25 monoclonal antibody,the level of CD4+CD25+ regulatory T cells was detected by flow cytometry,and the clinical data of APACHE Ⅱ scores of septic patients was considered in 24 hours.The correlation between level of CD4+CD25+ regulatory T cells in peripheral blood and APACHE Ⅱ scores in septic patients was analyzed.Results Compared with the healthy controls [(5.48±0.98)%],the level of CD4+ CD25+ regulatory T cells in sepsis group(10.31±2.32)%,severe sepsis group(14.27 43.33)%,septic shock group(15.32±3.98)% had a significant increase(P<0.05 or<0.01).The log value of regulatory T cells in each group correlated positively with the APACHE Ⅱ scores(r=0.829,P=0.032;r=0.868,P=0.021;r=0.913,P=0.009),and the total coefficient of correlation was 0.903(P=0.013).Conclusion The level of CD4+CD25+ regulatory T cells in peripheral blood in septic patients has an abnormal increase,and their levels are related with the severity of disease.
3.Effects of feixin decoction on the contents of hypoxia-inducible factor-1alpha and vascular endothelial growth factor in the rat model of hypoxic pulmonary hypertension.
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(5):676-680
OBJECTIVETo explore the effects of Feixin Decoction (FXD) on the hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in the rat model of hypoxic pulmonary hypertension (HPH), and to study its mechanisms for treating HPH.
METHODSForty healthy male SD rats were randomly divided into four groups, i. e., the normal control group, the HPH model group, the FXD group, and the Nifedipine group, 10 rats in each group. The HPH rat model was prepared using normal pressure intermittent hypoxia method. Except the normal control group, rats in the rest groups were fed in a self-made hypoxic plexiglass cabin, with the poor oxygen condition for 8 h daily for 14 successive days. Then the distilled water (at 30 mL/kg) was given by gastrogavage to rats in the normal control group and the HPH model group. FXD (at 28 g/kg) and Nifedipine (at 20 mg/kg) were given by gastrogavage to rats in the FXD group and the Nifedipine group respectively, once daily, for 14 successive days. Besides, hypoxia was continued for 14 days while medicating. The mean pulmonary artery pressure (mPAP) was detected on the second day after the last medication. The morphology of the pulmonary arteriole was detected. The ratio of pulmonary artery wall area and tube area (WA%) was determined. The protein and mRNA expressions of HIF-1alpha and VEGF were detected using immunohistochemistry and in situ hybridization technique.
RESULTSCompared with the normal control group, mPAP, WA%, and the protein and mRNA expressions of HIF-1alpha and VEGF significantly increased in the model group (P < 0.01, P < 0.05). Compared with the HPH model group, mPAP, WA%, and the protein and mRNA expressions of HIF-1alpha and VEGF significantly decreased in the FXD group (P < 0.01, P < 0.05).
CONCLUSIONSFXD down-regulated the expression of VEGF through decreasing the expression of HIF-1alpha. One of its mechanisms for treating HPH might be partially due to reversing the remodeling of pulmonary vascular smooth muscle.
Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypertension, Pulmonary ; drug therapy ; etiology ; metabolism ; Hypoxia ; complications ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism
4.The preventative effects of protein tyrosine kinase on the inflammation and airway remondeling in lung of guinea pigs with bronchial asthma.
Yun-fu ZHU ; Ai-guo DAI ; Rui-cheng HU
Chinese Journal of Applied Physiology 2009;25(2):151-154
AIMTo investigate the effects of protein tyrosine kinase on the inflammation and airway remodeling in lung of guinea pigs with bronchial asthma.
METHODS30 adult male guinea pigs were randomly divided into 3 groups (n=3): control group (C group), asthmatic group(A group)and genistein group (B group). Asthmatic model was established by ovalbumin intraperitoneal injection and ovalbumin inhalation. The total cell and the proportion of inflammatory cell in bronchial alveolar lavage fluid(BALF), inflammatory cell infiltration and index of remodeling of bronchiole were measured, respectively. The expression of p-tyrosine in lung tissue was examined by immunohistochemistry.
RESULTSThe total cell and proportion of eosinophil in BALF of A group were significantly higher than that of C group (P < 0.01), but compared with A group, the total cell and proportion of eosinophil in BALF of B group were much lower (P < 0.01). The number of eosinophile and lymphocyte of bronchiole in A group were significantly higher than that of C group (P < 0.01), but compared with A group, the number of eosinophile and lymphocyte in bronchiole of B group were much lower (P < 0.01). Compared with A group, the remodeling of bronchiole of B group was significantly relieved (P <0.01), there was no difference between B and C group (P > 0.05). Immunohistochemistry indicated that in A group the p-tyrosine was more positively expressed at the bronchial smooth muscle, bronchial epithelium, smooth muscle of vessel and inflammatory cell, especially at smooth muscle of bronchi and vessel and inflammatory cell than that of C group (P <0.01), there was no difference between B group and C group (P > 0.05).
CONCLUSIONPTK played a key role in inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma. The Protein tyrosine kinase inhibitor genistein could prevent and inhibit the inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma.
Airway Remodeling ; physiology ; Animals ; Asthma ; physiopathology ; prevention & control ; Genistein ; pharmacology ; Guinea Pigs ; Inflammation ; prevention & control ; Male ; Ovalbumin ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; physiology ; Random Allocation
5.Hypoxia-inducible factor-1 alpha regulates the role of vascular endothelial growth factor on pulmonary arteries of rats with hypoxia-induced pulmonary hypertension.
Chinese Medical Journal 2004;117(7):1023-1028
BACKGROUNDHypoxia-inducible factor-1alpha (HIF-1alpha) is one of the pivotal mediators in the response of lungs to decreased oxygen availability, and increasingly has been implicated in the pathogenesis of pulmonary hypertension. Vascular endothelial growth factor (VEGF), a downstream target gene of HIF-1alpha, plays an important role in the pathogenesis of hypoxic pulmonary hypertension and hypoxic pulmonary artery remodelling. In this study, we investigated the dynamic expression of HIF-1alpha and VEGF in pulmonary artery of rats with hypoxia-induced pulmonary hypertension.
METHODSForty male Wistar rats were exposed to hypoxia for 0, 3, 7, 14 or 21 days. Mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were estimated. Lungs were inflated and fixed for in situ hybridisation and immunohistochemistry.
RESULTSmPAP values were significantly higher than the control values after 7days of hypoxia [(18.4 +/- 0.4) mmHg, P < 0.05]. RVHI developed significantly after 14 days of hypoxia. Expression of HIF-1alpha protein increased in pulmonary arterial tunica intima of all hypoxic rats. In pulmonary arterial tunica media, HIF-1alpha protein was markedly increased by day 3 (0.20 +/- 0.02, P < 0.05), reached the peak by day 7, then declined after day 14 of hypoxia. HIF-1alpha mRNA increased significantly after day 14 of hypoxia (0.20 +/- 0.02, P < 0.05). VEGF protein began to increase markedly after day 7 of hypoxia, reaching its peak around day 14 of hypoxia (0.15 +/- 0.02, P < 0.05). VEGF mRNA began to increase after day 7 of hypoxia, then remained more or less stable from day 7 onwards. VEGF mRNA is located mainly in tunica intima and tunica media, whereas VEGF protein is located predominantly in tunica intima. Linear analysis showed that HIF-1alpha mRNA, VEGF and mPAP were correlated with hypoxic pulmonary artery remodelling. HIF-1alpha mRNA was positively correlated with VEGF mRNA and protein (P < 0.01).
CONCLUSIONHIF-1alpha and VEGF are both involved in the pathogenesis of hypoxia-induced pulmonary hypertension in rats.
Animals ; Blood Pressure ; Chronic Disease ; Hypertension, Pulmonary ; etiology ; Hypertrophy, Right Ventricular ; etiology ; Hypoxia ; complications ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; Male ; Pulmonary Artery ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Transcription Factors ; genetics ; physiology ; Vascular Endothelial Growth Factor A ; genetics ; physiology
6.The pathway of PI3k/Akt-aPKCiota/zeta-Nrf2 regulating the expression of gamma-glutamylcysteine synthetase in the bronchial epithelial cells of rats.
Chinese Journal of Applied Physiology 2011;27(1):115-119
OBJECTIVETo observe the effect of the signal pathway of phosphoinositol-3-kinase (PI3K)/Akt-antypical protein kinase C(iotazeta) (aPKC(iotazeta))-Nuclear factor-E2 related factor (Nrf2) on gamma-glutamylcysteine synthetase (gamma-GCS) of the bronchial epithelial cells of rats after exposure to cigarette smoke extracts (CSE).
METHODSgamma-GCS, Nrf2, p-Akt and p-aPKC(iotazeta) proteins were semi-quantified by Western blot. gamma-GCS protein expression was assessed by immunocytochemistry. gamma-GCS mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Nrf2 protein was observed by immunofluorescence. The rate of the cells expressed p-Akt were analyzed by flow cytometry. GSH content and gamma-GCS activity were measured.
RESULTSGSH content, Nrf2 protein of nucleus, p-aPKC(iotazeta) protein, p-Akt protein and positive cells, gamma-GCS protein and mRNA and activity were significantly increased after exposure to CSE for 3 hours. aPK(iotazeta) inhibitor RO813220 significantly reduced the expression of p-aPKC(iotazeta) protein, gamma-GCS protein and mRNA and activity, but enhanced Nrf2 protein of cytoplasm expression, had no effect on p-Akt. p-Akt inhibitor LY294002 and RO813220 + LY294002 decreased p-aPKC(iotazeta) protein, p-Akt protein and positive cells, gamma-GCS protein and mRNA and activity expression, increased Nrf2 protein of cytoplasm expression. The correlation analysises demonstrated that there were a positive correlation between Nrf2 and gamma-GCS, p-Akt, p-aPKC(iotazeta), between p-Akt and Nrf2, p-aPKC(iotazeta), gamma-GCS, between p-aPKC(iotazeta) and Nrf2, p-Akt, gamma-GCS.
CONCLUSIONCSE might upregulate gamma-GCS expression through PI3K/Akt-aPKC(iotazeta)-Nrf2 signaling pathway in the bronchial epithelial cells of rats.
Animals ; Bronchi ; cytology ; enzymology ; Environmental Exposure ; adverse effects ; Epithelial Cells ; enzymology ; GA-Binding Protein Transcription Factor ; metabolism ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Isoenzymes ; metabolism ; Male ; Oncogene Protein v-akt ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Protein Kinase C ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tobacco Smoke Pollution ; adverse effects
7.Roles of NRF2 regulating gamma-glutamylcysteine synthetase in lung of rats with chronic obstructive pulmonary disease.
Lin CHEN ; Ai-Guo DAI ; Rui-Cheng HU
Chinese Journal of Applied Physiology 2008;24(3):339-342
AIMTo investigate the expression and relationship of gamma-glutamylcysteine synthetase (gamma-GCS) and NF-E2-related factor2 (NRR2) in lung of rat with chronic obstructive pulmonary disease (COPD)in order to elucidate the possible important role of gamma-GCS and NRF2 in pathogenesis of COPD.
METHODSThe rat COPD model was established by intratracheal instillation of lipopolysaccharide twice and exposed to cigarette smoke daily. The gamma-GCS activity was measured, the expression of gamma-GCS mRNA in lung was examined by in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of NRF2, gamma-GCS in lung were detected by immunohistochemical (IH) and Western blot respectively.
RESULTSThe gamma-GCS activity was higher in COPD group than that in control group. The expressions of gamma-GCS mRNA in COPD group was stronger than those in control group. ISH showed that the gamma-GCS mRNA was expressed in alveolar epithelium and bronchial smooth muscle cell in COPD. The protein expressions of NRF2, gamma-GCS were significantly higher than the control group. IH showed that NRF2, gamma-GCS proteins were expressed in alveolar and bronchial epithelium in the COPD group. There was a positive correlation between NRF2 and gamma-GCS and gamma-GCS mRNA.
CONCLUSIONNRF2 may play an important role in the mechanism of COPD oxidative stress vis up-regulation of gamma-GCS.
Animals ; Glutamate-Cysteine Ligase ; metabolism ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; Pulmonary Disease, Chronic Obstructive ; metabolism ; physiopathology ; Random Allocation ; Rats ; Rats, Wistar
8.Location and expression of NF-kappaB in lung of rats with chronic obstructive pulmonary disease.
Shu-Dian LIN ; Ai-Guo DAI ; Shou-Min XI
Chinese Journal of Applied Physiology 2005;21(3):293-295
AIMTo elucidate the location and effects of transcription factor-nuclear factor-kappaB (NF-kappaB) in lung tissues of rats with chronic obstructive pulmonary disease (COPD).
METHODSFourteen male Wistar rats were randomly divided into COPD model and control groups equally. The COPD model was established by intratracheal instillation of lipopolysaccharide twice and exposure to cigarette smoke daily. We detected the NF-kappaB p65 protein in lung by immunohistochemical method, and the expression of NF-kappaB p65 mRNA in lung by in situ hybridization.
RESULTSImmunohistochemistry, the expression of NF-kappaB p65 protein in alveolar, bronchiolar epithelium and arteriolar endothelium was significantly higher in the COPD group (0.426 +/- 0.007, 0.434 +/- 0.012 and 0.313 +/- 0.007, respectively) than those of the control group (0.115 +/- 0.006, 0.116 +/- 0.005 and 0.095 +/- 0.007, respectively, all P < 0.01). In situ hybridization showed that the expressions of NF-kappaB p65 mRNA in alveolar epithelium (0.203 +/- 0.008), bronchiolar and arteriolar smooth muscle cell (0.208 + 0. 010 and 0.206 + 0.007) of rats in the COPD group were stronger than those in the control group (0.100 +/- 0.006, 0.102 +/- 0.002 and 0.103 +/- 0.003 respectively) by semiquantitative analysis (all P < 0.01).
CONCLUSIONThe expression and nuclear translocation of NF-kappaB may be the basis event of gene expression of many cytokines and inflammatory mediators, which may positively regulate gene expression of many cytokines and inflammatory mediators in various cell lines.
Animals ; Lung ; metabolism ; pathology ; Male ; Pulmonary Disease, Chronic Obstructive ; metabolism ; pathology ; Rats ; Rats, Wistar ; Transcription Factor RelA ; metabolism
9.Effects of adrenomedullin regulating inducible nitric oxide synthase on proliferation and apoptosis in hypoxic pulmonary artery smooth muscle cells.
Chi-guan LI ; Ai-guo DAI ; Cui-ping HUANG
Chinese Journal of Applied Physiology 2005;21(2):187-191
AIMTo investigate the effects of hypoxia on the proliferation and apoptosis of PASMC, to evaluate the role of iNOS protein expression and ADM on the hypoxic pulmonary hypertension (HPH) pathogenesis.
METHODSTo culture rat pulmonary artery smooth muscle cell (PASMC), cultured PASMC cells were grouped into: normoxic group; hypoxic group; hypoxia + L-NAME group; hypoxia+ ADM group. Proliferation of PASMC were investigated by MTT and PCNA. Apoptosis of PASMC were examined by flow-cytometry. Westen blot was used to measure protein expression of iNOS induced by hypoxia.
RESULTS(By MTT, the value of 24 h hypoxia was significantly higher than that in the normoxic group (P < 0.01), the value of the hypoxia + ADM was significantly lower than that in hypoxia group, the value of the hypoxia + L-NAME was significantly higher than those of hypoxic group and normoxic group (P < 0.01). (2) By immunohistochemistry, PCNA was poorly positive in PASMC, whereas positive after 24 h hypoxia (P < 0.01), ADM inhibited the expression of PCNA significantly (P < 0.01), whereas L-NAME increased the expression of PCNA significantly (P < 0.01). (3) By FCM, apoptosis index was not significantly different between the normoxic group, hypoxic group, hypoxia + L-NAME and hypoxia + ADM (P > 0.05). (4) By Western blot, iNOS expression was poorly positive in control group, positive after 4 h hypoxia (P < 0.01), increasing as the hypoxia environment continued (P < 0.01). L-NAME had no effect on iNOS protein, ADM promoted iNOS expression (P < 0.01).
CONCLUSION(1) Hypoxia stimulates the proliferation of PASMC, and has no obvious effects on the apoptosis of PASMC. (2) Hypoxia induces the expression of iNOS, ADM can increase expression of iNOS, ADM and INOS plays a role of protection in HPH pathogenesis.
Adrenomedullin ; pharmacology ; Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cell Proliferation ; drug effects ; Cells, Cultured ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; pathology ; Nitric Oxide Synthase Type II ; metabolism ; Pulmonary Artery ; cytology ; pathology ; Rats ; Rats, Sprague-Dawley
10.The expression and possible role of SENP1 in the pulmonary vascular wall of rat during the development of hypoxic pulmonary hypertension.
Hua TIAN ; Ai-Guo DAI ; Dai-Yan FU ; Rui-Cheng HU ; Li-Ming ZHU
Chinese Journal of Applied Physiology 2012;28(2):123-127
OBJECTIVETo investigate the dynamic expression and role of SENP1 (SUMO-specific proteases-1) in the pulmonary vascular wall of rat during the development of hypoxic pulmonary hypertension (HPH).
METHODSForty adult male Wistar rats were randomly divided into 5 groups (n = 8), and exposed to normoxia (Control group) or exposed to hypoxia for 3, 7, 14 or 21 d, respectively. The HPH models were established by normobaric intermittent hypoxia. Mean pulmonary arterial pressure (mPAP), right ventricle hypertrophy index (RVHI), and vessel morphometry were measured. Reverse transcriptase-polymerase chain reaction(RT-PCR) and in situ hybridization were used to determine the mRNA expression of SENP1. Immunohistochemistry and Western blot were used to determine the protein expression of SENP1.
RESULTSThe hypoxic rats developed pulmonary vascular remodeling in pulmonary arterioles after 7 d of hypoxia exposure. Pulmonary vascular remodeling in pulmonary arterioles significantly increased after 14 d of hypoxia. The level of mPAP in hypoxic rats increased significantly after 7 d of hypoxia, reached its peak after 14 d of hypoxic exposure. RVHI was markedly increased after 14 d of hypoxia. In situ hybridization and immunohistochemical analysis showed that SENP1 mRNA and protein were positively stained in control. SENP1 mRNA expression had little changes after exposure to hypoxia compared with the control, however, SENP1 protein expression was declined gradually after 7 d of hypoxia. The results of RT-PCR and Western blot showed that the same dynamic expression of SENP1 mRNA and protein in lung tissues of rats. Linear correlation analysis showed that SENP1 protein were negatively correlated with mPAP, pulmonary vascular remodeling index and RVHI.
CONCLUSIONUnder chronic hypoxia, SENP1 protein can be degradated. The dynamic expression of SENP1 protein may play a role in implicating in the development of HPH.
Animals ; Endopeptidases ; metabolism ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Male ; Pulmonary Artery ; metabolism ; Rats ; Rats, Wistar