Cell Line ; Glucocorticoids ; pharmacology ; HSP90 Heat-Shock Proteins ; metabolism ; Humans ; Mucin 5AC ; metabolism ; Respiratory System ; drug effects ; metabolism

C1q nephropathy is a rare type of glomerulonephritis manifested as the deposition of C1q in the glomerular mesangium during immunofluorescent staining. Systemic lupus erythematosus and type I membranoproliferative glomerulonephropathy need to be excluded in the diagnosis of C1q nephropathy. C1q nephropathy has various manifestations under a light microscope, mainly including minimal change disease, focal segmental glomerulosclerosis, and proliferative glomerulonephritis. This disease is mainly manifested as persistent proteinuria or nephrotic syndrome and occurs more frequently in boys. Currently, glucocorticoids are mainly used for the treatment of this disease. Patients with C1q nephropathy show a good response to immunosuppressant treatment, but have a high rate of glucocorticoid resistance. Therefore, in this case, methylprednisolone pulse therapy or a combination with immunosuppressant treatment helps to achieve a good prognosis.
Complement C1q ; metabolism ; Diagnosis, Differential ; Glomerulonephritis ; diagnosis ; drug therapy ; etiology ; Glucocorticoids ; therapeutic use ; Humans ; Prognosis

Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine D3 are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucoccorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin D3, 1,25-dihydroxycholecalciferol [1.25-(OH)2D3], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin D3 and dexamethasone, one of glucocorticoids, on osteblastic cell line(MC3T#-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nm/ml of 1,25-(OH)2D3 and/or 10nM/ml, 100nM/ml, 1micrometer/ml of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with 1micrometer/ml of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in 1micrometer/ml of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin D3 was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin D3 at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin D3 at 2-day cultivation, and was again decreased at 3-day cultivation.
Bone and Bones ; Bone Resorption ; Calcitriol ; Cholecalciferol ; Collagen Type I ; Dexamethasone* ; Glucocorticoids ; Metabolism ; Osteoblasts* ; Osteogenesis ; Vitamins*

Chronic psychological stress can promote vascular diseases, such as hypertension and atherosclerosis. This study aims to explore the effects and mechanism of chronic psychological stress on aortic medial calcification (AMC). Rat arterial calcification model was established by nicotine gavage in combination with vitamin D₃ (VitD₃) intramuscular injection, and rat model of chronic psychological stress was induced by humid environment. Aortic calcification in rats was evaluated by using Alizarin red staining, aortic calcium content detection, and alkaline phosphatase (ALP) activity assay. The expression levels of the related proteins, including vascular smooth muscle cells (VSMCs) contractile phenotype marker SM22α, osteoblast-like phenotype marker RUNX2, and endoplasmic reticulum stress (ERS) markers (GRP78 and CHOP), were determined by Western blot. The results showed that chronic psychological stress alone induced AMC in rats, further aggravated AMC induced by nicotine in combination with VitD₃, promoted the osteoblast-like phenotype transformation of VSMCs and aortic ERS activation, and significantly increased the plasma cortisol levels. The 11β-hydroxylase inhibitor metyrapone effectively reduced chronic psychological stress-induced plasma cortisol levels and ameliorated AMC and aortic ERS in chronic psychological stress model rats. Conversely, the glucocorticoid receptor agonist dexamethasone induced AMC, promoted AMC induced by nicotine combined with VitD₃, and further activated aortic ERS. The above effects of dexamethasone could be inhibited by ERS inhibitor 4-phenylbutyrate. These results suggest that chronic psychological stress can lead to the occurrence and development of AMC by promoting glucocorticoid synthesis, which may provide new strategies and targets for the prevention and control of AMC.
Rats ; Animals ; Glucocorticoids/metabolism* ; Rats, Sprague-Dawley ; Nicotine/metabolism* ; Hydrocortisone/metabolism* ; Muscle, Smooth, Vascular ; Dexamethasone/metabolism* ; Vascular Calcification/metabolism* ; Myocytes, Smooth Muscle/metabolism* ; Cells, Cultured

OBJECTIVEEosinophils play a pivotal role in asthmatic airway inflammation. We previously found a significantly high expression of Slingshot-1L (SSH-1L) in peripheral eosinophils in acute exacerbations of asthma. Objective To investigate the expression and localization patterns of SSH-1L in peripheral blood eosinophils of asthmatic patients and their changes after treatment with inhaled corticosteroids.

METHODSWe recruited 4 outpatients with acute exacerbations of asthma who received no previous corticosteroid treatment and 1 healthy volunteer. From all the subjects 30 ml peripheral venous blood samples were collected before and after a 3-month treatment with inhaled fluticasone. The eosinophils were isolated, purified and counted, and the expressions of SSH-1L in the eosinophils were examined by RT-PCR and Western blotting. The localization of SSH-1L phosphatases in the peripheral eosinophils was detected by immunofluorescence assay in one patient.

RESULTSSSH-1L phosphatases distributed diffusely in the cytoplasm, especially dense near the membrane of the peripheral eosinophils. Glucocorticoids treatment resulted in a significant reduction in both the SSH-1L mRNA expression (0.7403∓0.1124 vs 0.4101∓0.0363, P=0.001) and SSH-1L protein expression (0.3410∓0.1337 vs 0.1543∓0.0551, P=0.039).

CONCLUSIONA high expression of SSH-1L in peripheral eosinophils in acute exacerbations of asthma may play a role in the activation and migration of eosinophils. The efficacy of inhaled corticosteroids in asthma control might be partly attributed to a down-regulated expression of SSH-1L.

Adult ; Aged ; Asthma ; blood ; drug therapy ; Eosinophils ; metabolism ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Middle Aged ; Phosphoprotein Phosphatases ; metabolism

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

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 ; Cell Line ; Dexamethasone ; pharmacology ; Glucocorticoids ; pharmacology ; Male ; Mesangial Cells ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; genetics ; metabolism ; Transfection

OBJECTIVETo explore the expression and significance of secretions of hypothalamus-pituitary-adrenal (HPA) axis in human hypertrophic scar.

METHODSHypertrophic scar tissues obtained from 12 patients with deep-partial thickness burn or full-thickness burn and normal skin tissues from the same 7 patients with hypertrophic scar were harvested for determination of gene expression of corticotrophin-releasing hormone (CRH), CRH receptor 1 (CRH-R1), pro-opiomelanocortin (POMC), melanocortin receptor 2 (MC-2R), and glucocorticoid receptor α (GR-α) by real-time fluorescence quantitative PCR. After addition of corresponding antibodies, distribution differences of CRH, CRH-R1, adrenocorticotropic hormone (ATCH), MC-2R, and GR-α were observed with immunohistochemical staining. Data were processed with t test.

RESULTSThe mRNA expression of CRH, CRH-R1, POMC, and GR-α in hypertrophic scar was respectively 3.1 ± 0.8, 0.05 ± 0.03, 0.020 ± 0.007, and 0.0030 ± 0.0010, which were significantly lower than those in normal skin (20.6 ± 4.7, 0.30 ± 0.12, 0.060 ± 0.020, and 0.0200 ± 0.0070, with t values from 2.10 to 4.75, P values all below 0.05). There was no statistical difference in MC-2R mRNA expression between hypertrophic scar and normal skin (t = 1.48, P = 0.15). Immunohistochemical observation showed CRH, CRH-R1, ACTH, MC-2R, and GR-α in hypertrophic scar were located in basal layer of epidermis, fibroblast of dermis, and tube wall of sweat gland. Expressions of these indexes could also be observed in sebaceous gland and hair follicle besides above-mentioned structures.

CONCLUSIONSDecreasing expression of active material of HPA axis may be related to formation of hypertrophic scar.

Adolescent ; Adrenocorticotropic Hormone ; metabolism ; Adult ; Child ; Cicatrix, Hypertrophic ; metabolism ; Female ; Glucocorticoids ; metabolism ; Humans ; Hypothalamo-Hypophyseal System ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Young Adult

OBJECTIVETo investigate the expression of CLCA3 and Muc5ac in nasal mucosa in allergic rhinitis rats and the effects of glucocorticoid on its expression.

METHODSThirty SD rats were randomly divided into allergic rhinitis group, dexamethasone group and control group. Expression of CLCA3 mRNA and Muc5ac protein in nasal mucosa were detected by RT-PCR and immunohistochemical assay, respectively.

RESULTSCLCA3 mRNA and Muc5ac protein in allergic rhinitis group were significantly higher than those in control group (t = 8.565, 5.317, P < 0.01, respectively). The increased expression of CLCA3 mRNA in allergic rhinitis group was well correlated with the expression of Muc5ac protein and the correlation coefficient was 0.813 (P < 0.05). After treatment with dexamethasone, the expression of CLCA3 mRNA and Muc5ac protein was notably lower than that in allergic rhinitis group (t = 3.102, 2.226, P < 0.05, respectively).

CONCLUSIONSThe stronger gene expression of CLCA3 exists, complicated with mucus overproduction in the nasal mucosa of allergic rhinitis rats. CLCA3 expression may play a pivotal role in mucus overproduction in allergic rhinitis. Dexamethasone substantially downregulates the expression of CLCA3 mRNA and Muc5ac protein.

Animals ; Calcium Channel Agonists ; metabolism ; Chloride Channels ; metabolism ; Dexamethasone ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Mucin 5AC ; metabolism ; Nasal Mucosa ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic, Perennial ; metabolism

OBJECTIVETo study the effects of glucocorticoid on skeletal muscle protein metabolism in burn sepsis and its possible mechanism.

METHODSThe rats were randomly divided into four groups with 15 rats in each group. Group B, 30% TBSA full-thickness burn was produced on the back and endotoxin (6 mg/kg bw) was given intraperitoneally after the injury to simulate burn sepsis. Groups C and D, glucocorticoid receptor antagonist RU38486 (10 mg/kg bw) was given by gavage 2 hours before or 2 hours after burn with endotoxin, respectively. Group A, the rats received only normal saline in same volume as endotoxin. Plasma levels of cortisol were determined with standard procedure. Extensor digitorium longus muscles (EDL) were procured from both legs 12 hours after the injury. After weighing, the proteolytic rate was determined in vitro in an incubation system with oxygen rich environment by high performance liquid chromatography. The gene expressions of ubiquitin, E(2)-14kDa and C2 in the muscles were determined by Northern blot analysis.

RESULTSThe weight of EDL was significantly lower in group B than in group A (t = 9.03, P < 0.01). Although the weight of EDL muscles was also lower in groups C and D than in group A, it was significantly higher than in group B (t = 2.26, 6.42, P < 0.05 or P < 0.01). The concentrations of plasma cortisol were markedly higher in groups B, C and D than in group A (t = 9.03 - 22.94, P < 0.01). A 58.8% (210/357) of the total and 335.5% (4.16/1.24) of myofibrillar proteolytic rate in group B was higher than in group A (t = 36.99 and t = 46.19, P < 0.01), respectively. The total and myofibrillar proteolytic rate in group D was 28.3% (161/567) and 49.6% (2.68/5.40) and in group C 18.9% (108/567) and 23.2% (1.25/5.40), which were lower than those in group B (t = 5.34 approximately 34.68, P < 0.01), respectively. Although the expressions of ubiquitin mRNA (2.4 kb), E(2)-14 kDa mRNA (1.2 kb) and C2 mRNA in groups C and D were significantly higher than in group A, all the values were lower than those in group B (t = 3.22, 11.32, P < 0.01), especially in group C.

CONCLUSIONSThe proteolytic rate of skeletal muscle, especially the myofibrillar proteolytic rate, was enhanced during burn with sepsis. Hypersecretion of glucocorticoid could upgrade the gene expression of ubiquitin system, resulting in hyperdegradation of skeletal muscle protein during burn with sepsis. Glucocorticoid receptor antagonist RU38486 could decrease the hyperdegradation of skeletal muscle during burn with sepsis.

Animals ; Burns ; metabolism ; Gene Expression Regulation ; Glucocorticoids ; physiology ; Hydrocortisone ; blood ; Male ; Mifepristone ; pharmacology ; Muscle Proteins ; metabolism ; Muscle, Skeletal ; metabolism ; Rats ; Rats, Wistar ; Sepsis ; metabolism ; Ubiquitin ; metabolism