1.Correlation between endotoxin tolerance in human monocyte leukemia cell line THP-1 with glucocorticoid receptor-alpha.
Zhaohui, YANG ; Xiangjun, BAI ; Haiping, WANG ; Zhanfei, LI ; Siqi, LI ; Bo, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(1):28-30
Human monocyte leukemia cell line THP-1 was stimulated with lipopolysaccharide (LPS) to simulate the sepsis model and the expression of human glucocorticoid receptor-alpha (GR-alpha) mRNA in montocytes with endotoxin tolerance was investigated. THP-1 cells were cultured in serum-free medium, randomly divided into groups A, B, C, D and E, and stimulated with 0, 10, 10, 100, 0 ng/mL LPS for 24 h followed with 100, 100, 10, 100, 0 ng/mL LPS for another 24 h respectively. The expression of GR-alpha mRNA was detected by semi-quantitative reverse transcriptional polymerase chain reaction. Tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme linked immunosorbent assay (ELISA). The results showed that the A values of GR-alpha/beta-actin in groups A, B, C, D and E was 0.607 +/- 0.006, 0.368 +/- 0.005, 0.484 +/- 0.008, 0.509 +/- 0.004 and 0.564 +/- 0.014 respectively with the difference being significant among the groups (P < 0.05). The GR-alpha mRNA expression was negatively correlated with the TNF-alpha expression (P < 0.01). It was concluded that the down-regulation of the expression of GR-alpha mRNA in endotoxin tolerance THP-1 cells might play an important role in the development of endotoxin tolerance in THP-1 cells.
Drug Tolerance
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Endotoxins/*pharmacology
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Leukemia, Monocytic, Acute/*metabolism
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Leukemia, Monocytic, Acute/pathology
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Monocytes/*metabolism
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RNA, Messenger/biosynthesis
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RNA, Messenger/genetics
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Receptors, Glucocorticoid/*biosynthesis
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Receptors, Glucocorticoid/genetics
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Tumor Cells, Cultured
2.Glucocorticoid pathway mediated the inhibition of testosterone in rats exposed to dibutyl phthalate.
Xiao-feng ZHANG ; Jing ZHENG ; Zi LI ; Yang ZHANG
Chinese Journal of Preventive Medicine 2009;43(8):710-713
OBJECTIVETo investigate the inhibitory mechanisms of testosterone (T) biosynthesis in rats exposed to dibutyl phthalate (DBP).
METHODSMale Wistar rats were randomly divided into five groups by weight, including 0.25, 0.50, 1.00, 2.00 g/kg DBP groups and corn oil control group, with 16 rats in each group. DBP was administered by gavage once a day. After 30 days exposure, eight rats in each group were killed and the others were killed after 15 days without DBP administration. The levels of T and glucocorticoid (GC) in serum were determined by radioimmunoassay. The expression levels of 11 beta-dedroxysteriod dehydrogenase (11 beta-HSD) mRNA and steroidogenesis acute regulatory protein (StAR) mRNA were determined by RT-PCR. The protein expression level of glucocorticoid receptor (GR) was investigated by Western blotting.
RESULTSDuring exposure period, in 1.00 and 2.00 g/kg DBP groups, the levels of T were (0.260 +/- 0.218) ng/ml and (0.260 +/- 0.342) ng/ml, the levels of GC were (13.470 +/- 5.661) ng/ml and (13.740 +/- 3.977) ng/ml, the levels of T and GC in control group were (1.045 +/- 1.222) ng/ml and (9.224 +/- 3.496) ng/ml. There were statistic differences between 1.00 and 2.00 g/kg DBP groups and control group (t(T) values were -2.295 and -2.295, t(GC) values were 2.159 and 2.296, respectively, P < 0.05). The expression level of StAR mRNA was significantly down-regulated in 1.00 and 2.00 g/kg DBP groups, while StAR/beta-Actin values were 0.657 +/- 0.060 and 0.407 +/- 0.033, and compared to control group (0.871 +/- 0.081), there was statistic difference (t values were -3.707 and -8.037, P < 0.05). In 1.00 and 2.00 g/kg DBP groups, the expression of 11 beta-HSD mRNA and the expression of GR protein were increased in DBP dose-dependent manner, while 11 beta-HSD/beta-Actin values were 0.538 +/- 0.138 and 0.988 +/- 0.133, and GR/beta-Actin were 0.785 +/- 0.106 and 0.956 +/- 0.076, respectively. There were statistic difference, as compared to the controls (0.285 +/- 0.106 and 0.275 +/- 0.035) (t(11 beta-HSD/beta-Actin) values were 2.829 and 7.860, t(GR/beta-Actin) values were 8.064 and 10.77, respectively, P < 0.05).Linear correlation and regression revealed that there were positive correlation between DBP dose and the expression levels of 11 beta-HSD mRNA and GR protein, with r values of 0.766 and 0.790, respectively. In post-exposure period, there were no statistic differences of all above index among DBP groups and control group.
CONCLUSIONDBP might inhibit T production in rats through GR mediation.
Animals ; Dibutyl Phthalate ; toxicity ; Glucocorticoids ; metabolism ; Male ; Rats ; Rats, Wistar ; Receptors, Glucocorticoid ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; biosynthesis ; blood
3.Glucocorticoid receptor agonist dexamethasone attenuates renal ischemia/reperfusion injury by up-regulating eNOS/iNOS.
Jiong ZHANG ; Jun-hua LI ; Le WANG ; Min HAN ; Fang XIAO ; Xiao-qin LAN ; Yue-qiang LI ; Gang XU ; Ying YAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):516-520
The aim of this study was to determine the effect of dexamethasone (DEX) on renal ischemia/reperfusion injury (IRI). C57BL/6 mice were randomly divided into Sham group, IRI group and DEX group. The mice in IRI and DEX groups subjected to renal ischemia for 60 min, were treated with saline or DEX (4 mg/kg, i.p.) 60 min prior to I/R. After 24 h of reperfusion, the renal function, renal pathological changes, activation of extracellular signal-regulated kinase (ERK) and glucocorticoid receptor (GR), and the levels of iNOS and eNOS were detected. The results showed DEX significantly decreased the damage to renal function and pathological changes after renal IRI. Pre-treatment with DEX reduced ERK activation and down-regulated the level of iNOS, whereas up-regulated the level of eNOS after renal IRI. DEX could further promote the activation of GR. These findings indicated GR activation confers preconditioning-like protection against acute IRI partially by up-regulating the ratio of eNOS/iNOS.
Animals
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Dexamethasone
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pharmacology
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Gene Expression Regulation, Enzymologic
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drug effects
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Glucocorticoids
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pharmacology
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Male
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Mice
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Nitric Oxide Synthase Type II
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biosynthesis
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Nitric Oxide Synthase Type III
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biosynthesis
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Receptors, Glucocorticoid
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agonists
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Reperfusion Injury
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enzymology
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pathology
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Up-Regulation
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drug effects
4.Increased Expression of Glucocorticoid Receptor beta Messenger RNA in Patients with Ankylosing Spondylitis.
Chang Keun LEE ; Eun Young LEE ; You Sook CHO ; Keun Ae MOON ; Bin YOO ; Hee Bom MOON
The Korean Journal of Internal Medicine 2005;20(2):146-151
BACKGROUND: Glucocorticoids have been known to be less effective for treating ankylosing spondylitis (AS) patients than for treating rheumatoid arthritis (RA) patients. To elucidate the mechanisms underlying this phenomenon, we evaluated whether the glucocorticoid receptor (GR) beta expression of the peripheral blood mononuclear cells (PBMCs) in patients with AS is increased compared with patients with RA. METHODS: PBMCs were isolated from the subjects of 3 study groups: the healthy controls (n=25), the RA patients (n=25), and the AS patients (n=25). All the subjects had never taken corticosteroids and the patients with RA or AS were newly diagnosed. The expression of GR beta messenger RNA (mRNA) was determined by reverse transcription of the total RNA, and this was followed by semi-quantitative polymerase chain reaction analysis (RT-PCR). RESULTS: The level of GR alpha mRNA expression was not different among three groups. GR beta mRNA expression of the AS patients (2.02 [range: 0.99-7.21], median [25th-75th percentiles]) was enhanced compared with that of the controls (0.78 [range: 0.43-1.62]) and the RA patients (0.98 [range: 0.79-1.18]). The level of GR beta mRNA expression was not related to the inflammatory markers or the disease activity score 28 for the RA patients, and it was not related to the Bath ankylosing spondylitis disease activity index for the AS patients. CONCLUSION: The expression of GR beta mRNA, which is a dominant negative regulator for the glucocorticoid response, was increased in AS patients. The results suggest that the increased expression of GR beta mRNA may be related to the ineffectiveness of glucocorticoids for the treatment of AS.
Adult
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Comparative Study
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Female
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*Gene Expression
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Genetic Markers
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Humans
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Leukocytes, Mononuclear/metabolism
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Male
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Middle Aged
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RNA, Messenger/biosynthesis/*genetics
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Receptors, Glucocorticoid/biosynthesis/*genetics
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Research Support, Non-U.S. Gov't
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Retrospective Studies
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Reverse Transcriptase Polymerase Chain Reaction
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Spondylitis, Ankylosing/*blood/genetics
5.Impact of psychosocial stress on airway inflammation and its mechanism in a murine model of allergic asthma.
Bei LI ; Xiao-Hong DUAN ; Jin-Feng WU ; Bao-Jun LIU ; Qing-Li LUO ; Hua-Liang JIN ; Yi-Jie DU ; Hong-Ying ZHANG ; Yu-Xue CAO ; Jing-Cheng DONG
Chinese Medical Journal 2013;126(2):325-334
BACKGROUNDIt has already been recognized that psychosocial stress evokes asthma exacerbation; however, the mechanism of how stress gets inside the body is not clear. This study aimed to observe the impact of psychosocial stress on airway inflammation and its mechanism in the ovalbumin-induced asthmatic mice combined with social disruption stress.
METHODSThirty-six male BALB/c mice were randomly divided into: control group, asthma group (ovalbumin-induced), asthma plus social disruption stress group (SDR), and SDR group. The open field video tracking system was used to assess animal behaviors. The invasive pulmonary resistance (RL) and dynamic lung compliance (cdyn) test system from Buxco was applied to detect pulmonary function. The enzyme-linked immunosorbent assay (ELISA) was utilized to determine OVA-IgE, T-helper type 2 (Th2) cytokines (IL-4, IL-5, IL-13) and corticosterone in mouse serum, the Th2 cytokines (IL-4, IL-5, IL-13, IL-6, TNF-α) in bronchoalveolar lavage fluid (BALF), and IL-6 and TNF-α levels in the supernatant of splenocytes cultured in vitro. Hematoxylin-eosin (H&E) staining was used to assess airway inflammation in lung histology. The cell count kit-8 assay (CCK-8) was applied to evaluate the inhibitory effect of corticosterone on splenocyte proliferation induced by lipopolysaccharide (LPS). Real time-PCR and Western blotting were utilized to determine glucocorticoid receptor (GR) mRNA and GR protein expression in lungs.
RESULTSThe open field test showed that combined allergen exposure and repeated stress significantly shortened the time the mice spent in the center of the open field (P < 0.01), increased ambulatory activity (P < 0.01) and the count of fecal boli (P < 0.01), but deceased vertical activity (P < 0.01). Results from pulmonary function demonstrated that airway hyperresponsiveness (AHR) was enhanced by psychosocial stress compared with allergy exposure alone. The ELISA results showed that cytokines in serum and BALF were significantly increased (P < 0.05). Moreover, the lung histology showed that infiltrated inflammatory cells were significantly increased in the asthma-SDR group compared with the asthma group (P < 0.05). Interestingly, serum corticosterone was remarkably raised by psychosocial stress (P < 0.05). In addition, the inhibitory effect of corticosterone on IL-6 and TNF-α in LPS-stimulated splenocyte cultures in vitro was diminished in the asthma-SDR group compared to the asthma group. The CCK-8 test revealed that the inhibition effect of corticosterone on splenocyte proliferation induced by LPS was significantly impaired in the SDR and asthma-SDR groups, while no significant effect was observed in the control and asthma groups. Furthermore, expression of GR mRNA and GR protein were significantly reduced in the lung tissues of the asthma-SDR group (P < 0.05).
CONCLUSIONSSocial disruption stress can promote anxiety behavior, activate the hypothalamic-pituitary-adrenal (HPA) axis, increase AHR and inflammation, and also impair glucocorticoid sensitivity and its function in a murine model of asthma. The down-regulation of GR expression induced by social disruption stress is in part associated with glucocorticoid insensitivity, which leads to asthma exacerbation.
Animals ; Anxiety ; etiology ; Asthma ; etiology ; Bronchial Hyperreactivity ; etiology ; Corticosterone ; blood ; Cytokines ; biosynthesis ; Disease Models, Animal ; Lung ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Glucocorticoid ; analysis ; physiology ; Stress, Psychological ; complications
6.Effects of jingui shenqi pill combined prednisone on expression of glucocorticoid receptor and its clinical effect in treating bullous pemphigoid patients.
Bao-guo LIU ; Zhi-ying LI ; Ming DU
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(10):881-884
OBJECTIVETo investigate the effect of Jingui Shenqi Pill (JSP) on the expression of glucocorticoid receptor (GR) in the skin lesion and its clinical effect in treating bullous pemphigoid (BP) patients.
METHODSThirty BP patients were randomly divided into the treatment group (n=15) treated with JSP plus prednisone and the prednisone group (n=15) with prednisone alone both for 4 weeks. And a normal control group was set up also. Expressions of GR-alpha and GR-beta in the skin lesion of BP patients and the normal skin of the normal control were detected by immunohistochemical assay. Results The total effective rate was 93.33% in the treatment group, significantly higher than that in the prednisone group which was 73.33% (P < 0.05); GR-alpha expression was higher in the treatment group than that in other two groups (P < 0.01), while GR-beta expression in the treatment group was lower than that in the prednisone group (P < 0.01).
CONCLUSIONJSP could increase GR-alpha expression and decrease GR-beta expression in the skin lesion of BP patients, so as to improve sensitivity of skin to glucocorticoid.
Aged ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Pemphigoid, Bullous ; drug therapy ; metabolism ; pathology ; Phytotherapy ; Prednisone ; therapeutic use ; Receptors, Glucocorticoid ; biosynthesis ; Skin ; drug effects ; metabolism ; pathology ; Treatment Outcome
7.Regulatory effect of ginsenoside on glucocorticoid receptor in mice with ischemic liver damage.
Ying-Lu FENG ; Bin-Bin CHENG ; Chang-Quan LING
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(3):252-254
OBJECTIVETo study whether ginsenoside (GS) can regulate the glucocorticoid receptor (GR) in mice with ischemic liver damage, and to preliminarily observe its dose-effect relationship for providing an experimental bases in seeking a new way to relieve the damage from view of GR.
METHODSAdult male SD mouse was used to establish liver ischemia model, and different doses (100, 50, and 25 mg/kg) of GS was given via gastric infusion before modeling. The maximal GR binding capacity (Bmax) of liver and the level of GR mRNA expression in liver were dynamically determined at various time points (2 h, 6 h, 12 h and 24 h) after modeling.
RESULTSCompared with the normal control group, GR Bmax and GR mRNA expression in model rats were lower obviously (P < 0.01). As compared with the control group, GR Bmax and GR mRNA expression in model rats treated with 50 mg/kg GS significantly raised at 2 h, 6 h, 12 h (P < 0.01), while the changes in modeling rats treated with other two doses of GS were of no statistical significance.
CONCLUSIONGS in dose of 50 mg/kg can elevate the GR Bmax of liver and the level of GR mRNA expression in liver of rats with ischemic damage.
Animals ; Gene Expression Regulation ; drug effects ; Ginsenosides ; pharmacology ; Ischemia ; physiopathology ; Liver ; blood supply ; drug effects ; metabolism ; Male ; Mice ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Receptors, Glucocorticoid ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors
8.Effects of different doses of hydrocortisone on acute lung injury in rats with early septic shock induced by Escherichia coli.
Tao ZHOU ; Xun-mei FAN ; Yong-qing WANG ; Yu-jie QI ; Hui CHEN ; Su-yun QIAN
Chinese Journal of Pediatrics 2004;42(9):644-648
OBJECTIVETo observe the effects of different doses of hydrocortisone (HC) on acute lung injury (ALI) and inflammatory response in rats at early stage of septic shock induced by Escherichia coli and to investigate the possible mechanisms for such differences.
METHODSALI model of early septic shock was induced in rats by two injections of Escherichia coli at 5 hours interval, with the first intraperitoneal injection of 6.50 x 10(10) cfu/kg and followed by an external jugular vein injection of 2.00 x 10(11) cfu/kg. Forty Wistar rats were randomly divided into the following five groups: normal control, ALI without HC treatment, high-dose HC (150 mg/kg), medium-dose HC (20 mg/kg) and low-dose HC (6 mg/kg). Two hours after the treatment, the specimens were collected for histopathological examination and the biological indexes of lung injury were measured. The expressions of intercellular adhesion molecule-1 (ICAM-1) and glucocorticoid receptor (GR) in lung tissues were also investigated by immunohistochemical assays.
RESULTSThe biological indexes of lung injury [wet/dry weight ratio (g/g), total protein concentration in bronchoalveolar lavage fluid (mg/L) and lung permeability index (10(-3))] in ALI group (4.76 +/- 0.10, 278.96 +/- 60.45, 4.73 +/- 0.60) were significantly increased as compared to those in normal control group (4.10 +/- 0.07, 67.46 +/- 13.27, 1.12 +/- 0.15) (P < 0.05). The grades of ALI pathologic changes in ALI group (11.13 +/- 1.13) was significantly higher than that in the normal control group (0.50 +/- 0.53, P < 0.05). The ratio of expression area of ICAM-1 in ALI group (0.149 +/- 0.037) was significantly increased as compared to that in the normal control group (0.051 +/- 0.018) (P < 0.05). The ratio of expression area of GR all group (0.043 +/- 0.037) was significantly decreased as compared to that in the normal Control group (0.124 +/- 9.040) (P < 0.05) After administration of HC, all the lung injury indexes, pathological grades and the ratios of expression area of ICAM-1 and GR were significantly improved, with the most remarkable effects observed in the low-dose HC group. The expressions of ICAM-1 and GR showed a significantly negative linear correlation (r = 0.55, P < 0.0001).
CONCLUSIONThese results indicated that the low-dose HC treatment had the most remarkable effects of improving the biological indexes of lung injury, inflammatory mediators and pathological changes. These HC dose dependent therapeutic effects might be associated with the level of GR expression.
Acute Lung Injury ; drug therapy ; metabolism ; microbiology ; Animals ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Escherichia coli ; Escherichia coli Infections ; complications ; Glucocorticoids ; therapeutic use ; Hydrocortisone ; therapeutic use ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Lung ; metabolism ; Rats ; Rats, Wistar ; Receptors, Glucocorticoid ; biosynthesis ; Shock, Septic ; drug therapy ; metabolism ; microbiology