2.Distribution of dexamethasone in cochlea after intratympanic injection in rats.
Yao-yao FU ; Tian-yu ZHANG ; Hui ZHAO ; Wen LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2009;44(3):237-241
OBJECTIVETo investigate the distribution and pharmacokinetics of dexamethasone of different concentrations in the inner ears of SD rats after intratympanic injection.
METHODSTotally 144 adult SD rats were anaesthetized and dexamethasone sodium phosphate of different concentrations (5 mg/ml, 10 mg/ml, 20 mg/ml) was injected into the tympanums. The rats were sacrificed at various postinjection survival times (5 min, 10 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 48 h, 72 h), and every 4 rats were included into each group. Then after a series of processes the inner ear tissue was cryostat sectioned. The distribution of dexamethasone was evaluated using immunofluorescence with semiquantitative analysis. Immunofluorescence was also used in another 4 normal SD rats to detect the distribution of Glucocorticoid receptor (GR) in the inner ear.
RESULTSDexamethasone was observed initially 15 min after local drug administration and 30 min to its peak level. The highest concentration of dexamethasone labeling was seen in the spiral ligament, organ of Corti and spiral ganglion, which paralleled the distribution of GR. The tissue concentration of 10 mg/ml and 20 mg/ml groups was higher than 5 mg/ml every corresponding time point, and the lasting time was also prolonged from 48 hours to 72 hours.
CONCLUSIONSDexamethasone can enter into the cochlear tissue quickly after transtympanic injection, and its distribution accords nearly exactly with that of GR. Increase of the concentration of dexamethasone results in higher tissue distribution and longer lasting time.
Animals ; Cochlea ; metabolism ; Dexamethasone ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; metabolism
3.Expression of glucocorticoid receptor alpha and beta in chronic rhinosinusitis with nasal polyp.
Ming-Ming WANG ; Hong-Ping ZHANG ; Zhe WANG ; Xue-Mei CHEN ; Da-Liang ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2007;42(2):143-146
OBJECTIVETo detect the expression of glucocorticoid receptor (GR) alpha and beta in nasal polyps, and analyze the possible relationship between over-expression of GR beta and steroid insensitivity.
METHODSThe expression of GR alpha and GR beta was examined by immunohistochemical SP method in the specimens from 17 patients with recurring nasal polyp, 18 patients with chronic rhinosinusitis and nasal polyp (CRSwNP), and 12 patients with chronic rhinosinusitis without nasal polyp (CRSsNP) that did not recur during follow-up for 1.5 - 2 years.
RESULTSThe difference of numbers of GR alpha-positive cells (x +/- s) between groups with recurrent nasal polyp (20.2 +/- 6.9), CRSwNP (20.7 +/-7.2) , CRSsNP (16.9 +/- 7.2) and normal subjects (16.1 +/- 5.3) was not significant (P > 0.05). The numbers of GR beta-positive cells in recurring group (34.2 +/- 7.4) or CRSwNP (31.5 +/- 5.9) were higher than that in CRSsNP (19.8 +/- 7.8) and normal group (10.1 +/- 6.7) respectively (all P < 0.05). There was a trend toward higher level in recurring polyp compared with that of CRSwNP patients without recurrence in follow-up period, although this was not statistically different (P = 0.558). The difference of GR beta/GR alpha ratios (x +/- s) in recurring specimens (1.80 +/- 0.47) and CRSwNP group (1.65 +/- 0.49) was significant compared with normal group (0.77 +/- 0.66) respectively (P < 0.05), while there was no significance compared with CRSsNP (1.23 +/- 0.27, P > 0.05).
CONCLUSIONSThe high expression of GR beta in nasal polyp is related to the development of nasal polyp.
Adult ; Aged ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Nasal Polyps ; metabolism ; Receptors, Glucocorticoid ; metabolism ; Rhinitis ; metabolism ; Sinusitis ; metabolism
4.Glucocorticoid receptor subunit expression in adenoid tissue of children with obstructive sleep apnea hypopnea syndrome.
Xiao-Wen ZHANG ; Yuan LI ; Min-Qiang XIE ; Ge-Hua ZHANG ; Xian LIU ; Zhen-Lin WANG ; Tao WANG ; Zhong-Han LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2007;42(2):109-112
OBJECTIVEThe expression of glucocorticoid receptor (GR) expression in obstructive sleep apnea hypopnea syndrome (OSAHS) in children is currently unknown. The aim of this study was to determine the GR-alpha and GR-beta status in the adenoidal tissues in children with OSAHS.
METHODSThirty-four pediatric patients (aged 3-14 years, median 7.8 years) had sleep study with polysomnography before adenoidectomy. According to the criteria of apnea hypopnea index (AHI) > or = 5 /h or/and apnea index (AI) > or = 1/h, they were divided into OSAHS and non-OSAHS sub groups. The study was based on fluorescent quantitative PCR (FQ-RT-PCR) for the mRNA expression of GR-alpha and GR-beta in the adenoidal tissues in children.
RESULTSGR isoforms mRNA encoding for expression of both GR-alpha and GR-beta were detected in the adenoids of all children. GR-alpha mRNA level [(9.40 +/- 3.06) x 10(5) cDNA copies/microg total RNA] in the adenoidal tissues in OSAHS was lower than those in the non-OSAHS [(1.60 +/- 0.26) x 10(6) cDNA copies/microg total RNA] (F = 40.285, P < 0.001), whereas no differences found for GR-beta [(1.57 +/- 0.35) x 10(4) cDNA copies/microg total RNA, (1.52 +/- 0.18) x 10(4) cDNA copies/microg total RNA]. GR-alpha/GR-beta ratio was 62.3 +/- 20. 3 in OSAHS and 107.4 +/- 24.4 in non-OSAHS. AHI or AI was not related to the mRNA levels of GR-alpha and GR-beta in OSAHS or non-OSAHS.
CONCLUSIONSGR-alpha and GR-beta were detectable in the adenoidal tissues in children. These data indicated that the relationship between the expressions of GR and the clinical significance in OSAHS need further and profound investigation.
Adenoids ; metabolism ; Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; RNA, Messenger ; genetics ; Receptors, Glucocorticoid ; metabolism ; Sleep Apnea, Obstructive ; metabolism
5.Role of glucocorticoid receptor and nuclear factor-kappaB in patients with stable chronic obstructive pulmonary disease.
Tao LIU ; Min PENG ; Bai-qiang CAI
Acta Academiae Medicinae Sinicae 2010;32(2):147-150
OBJECTIVETo explore the expressions of glucocorticoid receptor (GCR) and nuclear factor-kappaB (NF-kappaB) in patients with stable chronic obstructive pulmonary disease (COPD).
METHODSBronchial biopsies and blood specimens were obtained from 8 smokers with stable COPD (COPD group) and 8 nonsmokers with normal lung function (control group). The expressions of NF-kappaB and GCR in nucleus protein of bronchial biopsies and peripheral blood lymphocyte from these two groups were examined by Western blot.
RESULTSThe expressions of GCR in nuclear protein of peripheral blood lymphocyte and the bronchial biopsies, especially in bronchi, were significantly lower in COPD group than in control group (P<0.05, P <0.01). Also, the expressions of NF-kappaB in nuclear proteins of the bronchial biopsies and peripheral blood lymphocyte were significantly higher in COPD group than in control group (P<0.05). The expressions of NF-kappaB and GCR were significantly higher in bronchial biopsies than in peripheral blood lymphocyte in both groups (P<0.05, P<0.01).
CONCLUSIONChronic inflammation, especially airway inflammation, still exists in patients with stable COPD, as suggested by the different expressions of GCR and NF-kappaB between COPD patients and normal controls.
Aged ; Female ; Humans ; Male ; Middle Aged ; NF-kappa B ; metabolism ; Pulmonary Disease, Chronic Obstructive ; metabolism ; Receptors, Glucocorticoid ; metabolism
6.Effects of Chinese herbs on glucocorticoid receptor in rat brain regions with chronic immobilization stress.
Jia-Xu CHEN ; Jian-Xin YANG ; Xin ZHAO ; Zhu-Feng WANG ; Guang-Xin YUE
Chinese Journal of Applied Physiology 2005;21(4):402-406
AIMTo investigate the changes of glucocorticoid receptor (GR) in rat brain regions with chronic immobilization stress and the influence of Chinese herbs.
METHODSWe copied the rat model of chronic immobilization stress (180 min daily, repeated 7 days or 21 days), and characterized the changes of GR in hippocampus CA1, cortex, dentate gyrus via immunohistochemistry integrated image analysis.
RESULTSThe contents of glucocorticoid receptors in hippocampus CA1 , cortex, dentate gyrus were increased and the intensity of immunity reaction was significantly stronger in the model group of 7 days than that in the normal control group (P < 0.05). The contents of GR were significantly decreased and the intensity of immunity reaction was weaken in the model group of 21 days (P < 0.05).
CONCLUSIONCompared with the model group of 21 days, the contents of GR were significantly increased and immunity reaction was intensified in three groups of treatment with Chinese herbs (P < 0.05), in which the group treated with soothing-liver herbs Xiaoyaopowder was the best.
Animals ; Brain ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; drug effects ; metabolism ; Restraint, Physical ; Stress, Physiological
7.Effects of electromagnetic irradiation on glucocorticoid in serum and its receptor expression in rat hippocampus.
Mao-quan LI ; Yan-yan WANG ; Guang-bin ZHANG ; Zheng-ping YU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2007;25(4):215-219
OBJECTIVETo explore the role and mechanism of glucocorticoid (GC) in the harmful bio-effects of electromagnetic irradiation.
METHODSRats were exposed to 65 mW/cm(2) electromagnetic wave for 20 min. At 10 min, 30 min, 3 h, 12 h after irradiation, their learning and memory abilities were tested by Morris water maze. The levels of corticosterone (CORT) in serum were measured by radioimmunoprecipitation assay and the changes of total glucocorticoid receptor (GR) expression and GR nuclear translocation in rat hippocampus were measured by reverse transcription-polymerase chain reaction and Western blot.
RESULTSThe rats had learning and memory deficits at 10 min, 30 min and 3 h after irradiation, but at 12 h had no difference from the normal control. The levels of corticosterone in serum increased significantly at 10 min, 30 min, decreased at 3 h and increased significantly compared with 12 h after irradiation. GR mRNA and total GR protein expression in rat hippocampus had no significant changes at 10 min, 30 min after irradiation. At 3 h, 12 h GR mRNA expression significantly decreased by 69%, 76% respectively and GR total protein decreased by 58%, 67% respectively. There were significant differences between the two groups and the corresponding controls (P<0.05). And compared with the control, the GR nuclear translocation increased significantly at 3 h and 12 h (P<0.05).
CONCLUSIONGC may take part in the injury to learning and memory abilities after electromagnetic irradiation, and the non-genomic and genomic effects of GC may play a major role in the early and late stage, respectively.
Animals ; Corticosterone ; blood ; Electromagnetic Fields ; adverse effects ; Glucocorticoids ; blood ; Hippocampus ; metabolism ; radiation effects ; Male ; Rats ; Rats, Wistar ; Receptors, Glucocorticoid ; metabolism
8.Effect of glucocorticoid receptor beta on glucocorticoid action in glomerular mesangial cells.
Lei ZHANG ; Qing-nan HE ; Min ZHU ; Gang ZHOU ; Juan-juan DING ; Pin ZHOU ; Xiao-chuan WU ; Zhu-wen YI
Journal of Central South University(Medical Sciences) 2007;32(6):941-948
OBJECTIVE:
To construct mesangial cell lines over- or under- expressing glucocorticoid receptor beta (GRbeta), to investigate the effect of GRbeta on glucocorticoid biological function, and to determine whether the overexpression of GRbeta explains the glucocorticoid-resistant in glomerular mesangial cells (GMCs).
METHODS:
The recombinant human sense or anti-sense gene of GRbeta was transferred into the rat GMCs by retrovirus-mediated stable transfection technique. Expression of hGRbeta mRNA in GMCs was determined by reverse transcription of total RNA followed by quantitative reverse transcription-polymerase chain reaction (RT-PCR), and the product of RT-PCR was then analyzed by gene sequencing. The expression of hGRbeta protein in GMCs was tested by Western blot. The inhibitory rate of dexamethasone-mediated cells on lipopolysaccharide (LPS)-stimulated GMC proliferation was detected to assess the effect of GRbeta at different expression levels on the glucocorticoid action. The cell proliferative activity in different cells with different levels of GRbeta was tested by MTT chromatometry. The change of cell cycle was analyzed by flow cytometry.
RESULTS:
RT-PCR and gene sequencing showed that the recombinant sense and anti-sense genes were correctly integrated into genomic DNA of mesangial cells. The protein expression tested by Western blot showed that GRbeta in cells inserted with the sense hGRbeta gene was higher than those cells inserted with the anti-sense hGRbeta gene (109.74+/-10.63 vs. 19.08+/-1.01, P<0.05). The inhibitory rate of cell proliferation induced by dexamethasone was lower in GMCs transfected with sense hGRbeta gene than those transfected with anti-sense hGRbeta gene (18.47%+/-2.12% vs. 60.33%+/- 5.29%,P<0.05). Under the inhibition of dexamethasone, the decreased cell number of S-stage cells was significantly lower, and the increased cell number of G1- stage cells was significantly higher in GMCs transfected with sense hGRbeta gene than those of non-transfected cells.
CONCLUSION
The overexpression of GRbeta may inhibit the glucocorticoid action in GMCs. The GRbeta level in mesangial cells may be an important factor in determining whether they are sensitive or resistant to glucocorticoid.
Animals
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Cell Line
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Dexamethasone
;
pharmacology
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Glucocorticoids
;
pharmacology
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Male
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Mesangial Cells
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metabolism
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Rats
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Rats, Sprague-Dawley
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Receptors, Glucocorticoid
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genetics
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metabolism
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Transfection
9.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
10.Research progress on the expression and effect of glucocorticoid receptors in developing brain.
Chinese Journal of Contemporary Pediatrics 2007;9(3):278-280
Animals
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Brain
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growth & development
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metabolism
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Emotions
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Glucocorticoids
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therapeutic use
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Humans
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Memory
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RNA, Messenger
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analysis
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Receptors, Glucocorticoid
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analysis
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genetics
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physiology