DNA Mutational Analysis ; Drug Resistance ; genetics ; Genetic Variation ; Glucocorticoids ; pharmacology ; Humans ; Mutation ; Receptors, Glucocorticoid ; classification ; genetics ; physiology

Animals ; Brain ; growth & development ; metabolism ; Emotions ; Glucocorticoids ; therapeutic use ; Humans ; Memory ; RNA, Messenger ; analysis ; Receptors, Glucocorticoid ; analysis ; genetics ; physiology

OBJECTIVETo investigate the effets of flurothyl-induced neonatal recurrent seizures on glucocorticoid receptor (GR) expression in the rat brain.

METHODSForty-eight seven-day-old Sprague-Dawley rats were randomly divided into two groups: control and seizure. Seizures were induced by inhalant flurothyl daily for six consecutive days. Brains were sampled on postnatal days 13, 15 and 19. The expression of GR protein in the cerebral cortex was detected by Western blot and immunohistochemical method.

RESULTSThe expression of GR in the cerebral cortical plasma protein was significantly lower in the seizure group than in the control group on postnatal day 15. The expression of GR protein in the cerebral cortical nuclear protein decreased significantly in the seizure group compared with that in the control group on postnatal days 15 and 19 (p<0.05). Compared to the control group, the accumulated optical density (AOD) of GR immunoreactivity (IR) decreased significantly in the parietal cortex on postnatal day 13 (p<0.05), the AOD of GR IR decreased significantly in the parietal cortex and the temporal cortex on postnatal day 15 (p<0.05), and the AOD of GR IR decreased significantly in the parietal cortex, temporal cortex and the frontal cortex in the seizure group on postnatal day 19 (p<0.05).

CONCLUSIONSRecurrent seizures in neonatal rats result in abnormal GR expression in the cerebral cortex which might play an important role in short-term brain injury induced by early recurrent seizures.

Animals ; Blotting, Western ; Cerebral Cortex ; chemistry ; Female ; Hypothalamo-Hypophyseal System ; physiology ; Immunohistochemistry ; Male ; Pituitary-Adrenal System ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; analysis ; physiology ; Recurrence ; Seizures ; metabolism

BACKGROUNDGlucocorticoid (GC) insensitivity/GC resistance is an important etiological and prognostic factor in multiple diseases and pathophysiological processes such as scald, shock and asthma. The function of GC was mediated by glucocorticoid receptor (GR). Scald not only decreased the expression of GR but also reduced the affinity of GR, which played an important role in GC resistance in scalded rats. Whereas the molecular mechanism responsible for the decrease of GR affinity resulted from scald remains unclear. Recent studies showed that the changes of heat shock proteins (hsp) especially hsp90 and hsp70 of GR heterocomplex were associated with GR low affinity in vitro.

METHODSThe affinity of GR in hepatic cytosols and in the cytosols of SMMC-7721 cells were determined by radioligand binding assay and scatchard plot. GR heterocomplex in cytosols were captured by coimmunoprecipation and the levels of hsp90 and hsp70 of GR complex were detected by quantitative Western blotting.

RESULTSSimilar with that of hepatic cytosol of scalded rats, a remarkable decrease of GR affinity was also found in the cytosol of heat stressed SMMC-7721 cells. The level of hsp70 of GR complex in hepatic cytosol of scalded rats (30% total body surface area immersion scald) and in cytosol of heat stressed human hepatocarcinoma cell line SMMC-7721 were both increased by 1.5 fold, whereas no change of hsp90 in GR heterocomplex was found. According to the correlation analysis, there may be a positive relationship between increased hsp70 of GR complex and decreased GR affinity in the cytosols.

CONCLUSIONSThe primary results indicated that the level of hsp70 of GR heterocomplex was increased in the hepatic cytosol of scalded rats and the cytosol of heat stressed SMMC-7721 cells. The increase of hsp70 of GR complex might be associated with the decrease of GR affinity.

Animals ; Blotting, Western ; Burns ; physiopathology ; Cell Line ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Response ; physiology ; Immunoprecipitation ; Protein Binding ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; metabolism

OBJECTIVETo study the effects of different assemblages formed by components of Shengmai Powder (SMP) on glucocorticoid receptor (GR) in liver of thermal injured rat to find the optimal ratio of assembling for GR regulation.

METHODSWith a orthogonal design adopted, the dosage of each component of SMP, including the total saponins of Ginseng (G), the water extract of lilyturf root (L), and the water extract of schisandra fruit (S), was ranked in three levels, namely, no participating, low dosage (G 7.1 mg, L 17.2 mg, S 9.6 mg), high dosage (G 14.2 mg, L 34.4 mg, S 19.2 mg). The components were assigned by L9(3(4)) orthogonal table and grouped, the best assembling ratio was determined through direct and variance analysis.

RESULTSAfter being acted by the different assemblages, the maximum binding volume of GR in rat's liver cell suspension was 161 +/- 26 fmol/mg protein in group 1, 271 +/- 40 fmol/mg protein in group 2, 166 66 fmol/mg protein in group 3, 222 +/- 45 fmol/mg protein in group 4, 192 +/- 26 fmol/mg protein in group 5, 194 +/- 23 fmol/mg protein in group 6, 166 +/- 15 fmol/mg protein in group 7, 165 +/- 47 fmol/mg protein in group 8 and 211 +/- 79 fmol/mg protein in group 9. The optimal GR level appeared during the dosage of G, L and S was 7.1 mg, 17.2 mg and 19.2 mg, respectively.

CONCLUSIONThe best assembling ratio of SMP for regulating GR in the liver of thermal injured rat was G:L:S = 3:3:4.

Animals ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Hot Temperature ; adverse effects ; Liver ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; metabolism ; Stress, Physiological ; physiology

OBJECTIVETo investigate the role of the change in glucocorticoid receptor in stress-induced gastric ulcer in severely burned rats.

METHODSWistar rats inflicted with 30% TBSA full-thickness burn were employed as the model. The dynamic postburn change in the glucocorticoid receptor in gastric mucosal tissue and gastric mucosal injury (injury index) were observed.

RESULTSThe gastric mucosal cortisol content increased as of 3 postburn hours (PBHs) and peaked at 12 PBHs. But the cytoplasmic glucocorticoid receptor content in the gastric mucosal cells decreased evidently. In addition, the gastric mucosal injury index increased obviously at 12 and 24 PBHs.

CONCLUSIONThe decrease of gastric mucosal cytoplasmic glucocorticoid receptor content might be closely related to the stress-induced gastric mucosal injury.

Animals ; Burns ; complications ; metabolism ; Disease Models, Animal ; Male ; Rats ; Rats, Wistar ; Receptors, Glucocorticoid ; metabolism ; physiology ; Stomach Ulcer ; complications ; metabolism ; Stress, Physiological ; complications

OBJECTIVETo study the changes of glucocorticoid receptor (GR) expression in embryonic rat cortical neurons exposed to transient Mg(2+)-free treatment.

METHODSSix days after rat cortical neuronal cultures, two groups were created based on the medium to which were transiently exposed. The control group was exposed to a physiological solution (PS), and the Mg(2+)-free group was exposed to the same medium as the control group except for the removal of magnesium. The expression of GR mRNA and protein was determined by real-time PCR and immunocytochemistry staining 1, 7 and 12 days after transient Mg(2+)-free treatment.

RESULTSCompared to the control group, the Mg(2+)-free group displayed the significantly less accumulated optical density (AOD) of GR immunoreactivity 12 days after transient Mg(2+)-free treatment (p<0.05). On the contrary, GR mRNA expression increased significantly 1 and 7 days after transient Mg(2+)-free treatment in the Mg(2+)-free group (p<0.05).

CONCLUSIONSGR expression is modified following Mg-free-induced injury in cultured developing neurons in rats.

Animals ; Cell Survival ; Cells, Cultured ; Cerebral Cortex ; metabolism ; Fetus ; metabolism ; Hypothalamo-Hypophyseal System ; physiology ; Magnesium ; physiology ; Neurons ; metabolism ; Pituitary-Adrenal System ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Receptors, Glucocorticoid ; analysis ; genetics

BACKGROUND AND OBJECTIVE: Acute liver failure (ALF) is a type of disease with high mortality and rapid progression with no specific treatment methods currently available. Glucocorticoids exert beneficial clinical effects on therapy for ALF. However, the mechanism of this effect remains unclear and when to use glucocorticoids in patients with ALF is difficult to determine. The purpose of this study was to investigate the specific immunological mechanism of dexamethasone (Dex) on treatment of ALF induced by lipopolysaccharide (LPS)/D-galactosamine (D-GaIN) in mice. METHODS: Male C57BL/6 mice were given LPS and D-GaIN by intraperitoneal injection to establish an animal model of ALF. Dex was administrated to these mice and its therapeutic effect was observed. Hematoxylin and eosin (H&E) staining was used to determine liver pathology. Multicolor flow cytometry, cytometric bead array (CBA) method, and next-generation sequencing were performed to detect changes of messenger RNA (mRNA) in immune cells, cytokines, and Kupffer cells, respectively. RESULTS: A mouse model of ALF can be constructed successfully using LPS/D-GaIN, which causes a cytokine storm in early disease progression. Innate immune cells change markedly with progression of liver failure. Earlier use of Dex, at 0 h rather than 1 h, could significantly improve the progression of ALF induced by LPS/D-GaIN in mice. Numbers of innate immune cells, especially Kupffer cells and neutrophils, increased significantly in the Dex-treated group. In vivo experiments indicated that the therapeutic effect of Dex is exerted mainly via the glucocorticoid receptor (Gr). Sequencing of Kupffer cells revealed that Dex could increase mRNA transcription level of nuclear receptor subfamily 4 group A member 1 (Nr4a1), and that this effect disappeared after Gr inhibition. CONCLUSIONS In LPS/D-GaIN-induced ALF mice, early administration of Dex improved ALF by increasing the numbers of innate immune cells, especially Kupffer cells and neutrophils. Gr-dependent Nr4a1 upregulation in Kupffer cells may be an important ALF effect regulated by Dex in this process.
Animals ; Dexamethasone/therapeutic use* ; Disease Models, Animal ; Kupffer Cells/physiology* ; Liver Failure, Acute/pathology* ; Male ; Mice ; Mice, Inbred C57BL ; Nuclear Receptor Subfamily 4, Group A, Member 1/physiology* ; Receptors, Glucocorticoid/physiology*

Glucocorticoid receptor (GR) is identified as a member of nuclear receptor family. To exert its biological action, the ligand bound GR is translocated from the cytoplasm into the nucleus by regulating transcriptional signals of related genes. In clinical practice, the effects of glucocorticoid are often mediated by GR signaling pathways. An increasing number of studies have indicated that GR signaling pathways play an essential role in the proliferation, invasion and prognosis of bladder cancer. Meanwhile, the new-generation selective GR activator improves its anti-tumor effects, and at the same time reduces the adverse reactions of hormones, which probably raises the prospect for the treatment of bladder cancer.
Animals ; Antineoplastic Agents ; pharmacology ; Cell Nucleus ; genetics ; Humans ; Prognosis ; Protein Transport ; genetics ; Receptors, Glucocorticoid ; agonists ; physiology ; Signal Transduction ; genetics ; Transcriptional Activation ; drug effects ; physiology ; Urinary Bladder Neoplasms ; genetics ; physiopathology

Background: Glucocorticoid (GC) is the first-line therapy for asthma, but some asthmatics are insensitive to it. Glucocorticoid-induced transcript 1 gene (GLCCI1) is reported to be associated with GCs efficiency in asthmatics, while its exact mechanism remains unknown. Methods: A total of 30 asthmatic patients received fluticasone propionate for 12 weeks. Forced expiratory volume in 1 s (FEV) and GLCCI1 expression were detected. Asthma model was constructed in wild-type and GLCCI1 knockout (GLCCI1) mice. Glucocorticoid receptor (GR) and mitogen-activated protein kinase phosphatase 1 (MKP-1) expression were detected by polymerase chain reaction and Western blotting (WB). The phosphorylation of p38 mitogen-activated protein kinase (MAPK) was also detected by WB. Results: In asthmatic patients, the change of FEV was well positively correlated with change of GLCCI1 expression (r = 0.430, P = 0.022). In animal experiment, GR and MKP-1 mRNA levels were significantly decreased in asthmatic mice than in control mice (wild-type: GR: 0.769 vs. 1.000, P = 0.022; MKP-1: 0.493 vs. 1.000, P < 0.001. GLCCI1: GR: 0.629 vs. 1.645, P < 0.001; MKP-1: 0.377 vs. 2.146, P < 0.001). Hydroprednisone treatment significantly increased GR and MKP-1 mRNA expression levels than in asthmatic groups; however, GLCCI1 asthmatic mice had less improvement (wild-type: GR: 1.517 vs. 0.769, P = 0.023; MKP-1: 1.036 vs. 0.493, P = 0.003. GLCCI1: GR: 0.846 vs. 0.629, P = 0.116; MKP-1: 0.475 vs. 0.377, P = 0.388). GLCCI1 asthmatic mice had more obvious phosphorylation of p38 MAPK than wild-type asthmatic mice (9.060 vs. 3.484, P < 0.001). It was still higher even though after hydroprednisone treatment (6.440 vs. 2.630, P < 0.001). Conclusions: GLCCI1 deficiency in asthmatic mice inhibits the activation of GR and MKP-1 and leads to more obvious phosphorylation of p38 MAPK, leading to a decremental sensitivity to GCs. Trial Registration ChiCTR.org.cn, ChiCTR-RCC-13003634; http://www.chictr.org.cn/showproj.aspx?proj=5926.
Animals ; Asthma ; drug therapy ; metabolism ; Dual Specificity Phosphatase 1 ; genetics ; metabolism ; Forced Expiratory Volume ; genetics ; physiology ; Glucocorticoids ; therapeutic use ; Mice ; Mice, Knockout ; Phosphorylation ; genetics ; physiology ; Receptors, Glucocorticoid ; deficiency ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism