Glucocorticoid receptor

Resistance or sensitivity to glucocorticoids is considered to be of crucial importance for disease prognosis in childhood acute lymphoblastic leukemia. Prednisolone exerted a delayed biphasic effect on the resistant CCRF-CEM leukemic cell line, necrotic at low doses and apoptotic at higher doses. At low doses, prednisolone exerted a pre-dominant mitogenic effect despite its induction on total cell death, while at higher doses, prednisolone's mitogenic and cell death effects were counterbalanced. Early gene microarray analysis revealed notable differences in 40 genes. The mitogenic/biphasic effects of prednisolone are of clinical importance in the case of resistant leukemic cells. This approach might lead to the identification of gene candidates for future molecular drug targets in combination therapy with glucocorticoids, along with early markers for glucocorticoid resistance [1].

Nitric oxide is believed to participate in nonspecific cellular immunity. Gram negative bacterial endotoxins increase the production of reactive nitrogen intermediates (RNI) in phagocytic cells by inducing the enzyme nitric oxide synthase II (NOS II). Anti-inflammatory glucocorticoids attenuate endotoxin-induced increases in RNI. This study evaluated the effect of in vivo administration of prednisolone on Escherichia coli lipopolysaccharide endotoxin (LPS)-induced increases in plasma RNI and neutrophil mRNA for NOS II and production of RNI in the rat. We show that LPS rapidly induces mRNA for NOS II and production of RNI (NO2- and NO3- anion) in rat neutrophils within 2 hr after in vivo administration of a sublethal dose of 0.5 mg/kg, i.v. A pharmacologic dose of prednisolone (50 micrograms/kg, im) given 15 min before LPS-attenuated production of NO2- and NO3- by neutrophils and suppressed LPS-stimulated mRNA for NOS II. 3-Amino, 1,2,4-triazine inhibited NO2- and NO3- production without affecting gene expression for NOS II. These data demonstrate that LPS rapidly induces functional gene expression for NOS II and prednisolone prevents induction of NOS II activity by inhibiting transcription of its mRNA [2].

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Diaphragm atrophy and weakness occur after administration of massive doses of corticosteroids for short periods. In the present study the effects of prolonged administration of moderate doses of fluorinated and nonfluorinated steroids were investigated on contractile properties and histopathology of rat diaphragm. 60 rats received saline, 1.0 mg/kg triamcinolone, or 1.25 or 5 mg/kg i.m. prednisolone daily for 4 wk. Respiratory and peripheral muscle mass increased similarly in control and both prednisolone groups, whereas triamcinolone caused severe muscle wasting. Maximal tetanic tension averaged 2.23 +/- 0.54 kg/cm2 (SD) in the control group. An increased number of diaphragmatic bundles in the 5-mg/kg prednisolone group generated maximal tetanic tensions < 2.0 kg/cm2 (P < 0.05). In addition, fatigability during the force-frequency protocol was most pronounced in this group (P < 0.05). In contrast, triamcinolone caused a prolonged half-relaxation time and a leftward shift of the force-frequency curve (P < 0.05). Histological examination of the diaphragm showed a normal pattern in the control and 1.25-mg/kg prednisolone group. Myogenic changes, however, were found in the 5-mg/kg prednisolone group and, more pronounced, in the triamcinolone group. Selective type IIb fiber atrophy was found in the latter group, but not in the prednisolone groups. In conclusion, triamcinolone induced type IIb fiber atrophy, resulting in reduced respiratory muscle strength and a leftward shift of the force-frequency curve. In contrast, 5 mg/kg prednisolone caused alterations in diaphragmatic contractile properties and histological changes without fiber atrophy [3].

Prednisolone treatment [1]
Concentrations of Prednisolone were selected on the basis of the average in vivo dosage administrated intravenously to children at ages between 1 month and 12 years old (details in supplementary data, file: CCRFCEM Cytotoxixity Assay.xls). Also, bioactivity in cortisol equivalents is estimated to be in the range of 40–200 nM. To ensure that the study covers these ranges, prednisolone was diluted to the following 12 concentrations: control, 10 nM, 100 nM, 1 μM, 5.5 μM, 11 μM, 22 μM, 44 μM, 88 μM, 175 μM, 350 μM, and 700 μM.

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Cell proliferation assay [1]
Cell population counts were determined with the use of a NIHON KOHDEN CellTaq-α hematology analyzer. Cells were counted at the −24 h time point as well as 0 h, 4 h, 24 h, 48 h, and 72 h after initiation of exposure to prednisolone. For this purpose, 200 μl of cell suspensions were obtained from each flask and counted directly with the analyzer.

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Protein extraction and Western blotting [1]
Cells were harvested after 1 h and 4 h exposure to different concentrations of Prednisolone. Protein extraction and Western blotting were performed as previously described. Total protein content was determined by the Bradford method using bovine serum albumin as a standard. Proteins were separated by SDS-PAGE and Western blotting was carried out, with anti-p65 antibodies

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Microarray analysis [1]
cDNA microarray chips (1200 genes) were obtained from TAKARA (Human Cancer Chip v.40). Hybridization was performed with the CyScribe Post-Labeling kit as described by the manufacturer, utilizing the Cy3 and Cy5 fluorescent dyes. Slides were scanned with a microarray scanner. Images were generated with ScanArray microarray acquisition software. cDNAs from three experimental setups were used, each one consisting of three independent experiments. The experimental setups consisted of the three following pairs: control vs. 10 nM Prednisolone (designated as 0vs1), 10 nM prednisolone vs. 700 μM prednisolone (designated as 1vs3), control vs. 700 μM prednisolone (designated as 0vs3). This is a ‘simple loop’ experimental design, taking into account all possible combinations between samples, as previously described. Raw microarray data are available as supplementary data.

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Real-time reverse transcription PCR (qRT-PCR) [1]
The GRIM19 (NDUFA13) gene was tested from three samples control, 10 nM and 700 μM Prednisolone at 4 h and 48 h treatment, using the one-step Plexor™ qRT-PCR kit. The set of primers was designed using the on-line tool Plexor™ Primer Design System v1.2 by Promega

Study design, animals, and treatment [3]
60 adult male Wistar rats, aged 14 wk, weighing 350-400 g, were randomized in quadruplets, into one of four treatment groups: control (C), saline, 0.05 ml/d i.m.; low dose prednisolone (LD), 1.25 mg/kg per d i.m.; high dose prednisolone (HD), 5 mg/kg per d i.m.; or triamcinolone-diacetate (TR), 1 mg/kg per d i.m. Dilution of medication was performed such that with each injection all animals received a similar volume (0.05 ml). During 4 wk the animals were injected daily in the left hindlimb. They were fed ad libitum and weighed twice weekly. After the treatment period, contractile properties, histological, and histochemical characteristics of the diaphragm were examined.[3]

rabbit LD50 intravenous 360 mg/kg Chemical and Pharmaceutical Bulletin., 22(1439), 1974 [PMID:4373183]

[1]. Prednisolone exerts late mitogenic and biphasic effects on resistant acute lymphoblastic leukemia cells: Relation to early gene expression. Leuk Res, 2009. 33(12): p. 1684-95.

[2]. Rapid induction of messenger RNA for nitric oxide synthase II in rat neutrophils in vivo by endotoxin and its suppression by prednisolone. Proc Soc Exp Biol Med. 1994 Mar;205(3):220-9.

[3]. Triamcinolone and prednisolone affect contractile properties and histopathology of rat diaphragm differently. J Clin Invest. 1993 Sep;92(3):1534-42.

Prednisolone sodium phosphate can cause developmental toxicity according to state or federal government labeling requirements.
Prednisolone sodium phosphate is an organic sodium salt of prednisolone phosphate. It is an ophthalmology drug used for the treatment of short-term inflammatory eye conditions and helps relieve inflammation, redness and irritation of the eyes. It has a role as an ophthalmology drug, an anti-inflammatory agent, a glucocorticoid receptor agonist, a prodrug and an anti-allergic agent. It is an organic sodium salt and a glucocorticoid. It contains a prednisolone phosphate(2-).
Prednisolone Sodium Phosphate is the sodium phosphate salt form of prednisolone, a synthetic glucocorticoid with anti-inflammatory and immunomodulating properties. As a glucocorticoid receptor agonist, prednisolone sodium phosphate binds to specific intracellular glucocorticoid receptors, and causes the ligand- receptor complex to be translocated to the nucleus where it initiates the transcription of glucocorticoid-responsive genes such as various cytokines and lipocortins. Lipocortins inhibit phospholipase A2, thereby blocking the release of arachidonic acid from membrane phospholipids and preventing the synthesis of prostaglandins and leukotrienes, both potent mediators of inflammation. This agent also decreases the number of circulating lymphocytes, induces cell differentiation, and stimulates apoptosis in sensitive tumor cell populations.
See also: Prednisolone (has active moiety); Neomycin sulfate; prednisolone sodium phosphate (component of) ... View More ...

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In conclusion, this study shows: (a) prednisolone exerts a delayed biphasic effect on CCRF-CEM cells, necrotic at low doses and apoptotic at higher doses, (b) at low doses, prednisolone exerts a pre-dominant mitogenic effect despite its induction on total cell death, while at higher doses, prednisolone's mitogenic and cell death effects are counterbalanced. These mitogenic effects are of clinical importance in the case of resistant leukemic cells. It is of crucial importance if a certain administered dose of GCs to an ALL patient possesses proliferative rather than suppressive actions, (c) NF-κB is constitutively localized in the nucleus and its inhibition emerges as a possible candidate for the treatment of resistant leukemia, (d) prednisolone activates genes related to at least four different pathways upon 4 h treatment: apoptosis and tumor suppression, cell cycle progression, metabolism and intra- and extra-cellular signaling. Several of these genes manifested a biphasic differential expression profile. This approach might lead to the identification of genes candidates for future molecular drug targets in combination therapy with GCs. These drugs may affect the potential of GCs to inhibit growth of resistant leukemic cells. Also, this type of approach could identify potential early markers for GCs resistance. Early detection of resistance could facilitate the efficiency of ALL therapies.[1]

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In conclusion, the present study shows that triamcinolone and prednisolone have different effects on morphology and contractile properties of the rat diaphragm. Fluorinated steroids such as triamcinolone caused severe muscle wasting due to selective type Ilb fiber atrophy, resulting in markedly reduced respiratory muscle strength. Prednisolone in four times higher doses caused a tendency towards lower tetanic tensions and increased fatigability of diaphragmatic muscle bundles, with distinct alterations in muscle histology.[3]

C21H27NA2O8P

484.39

484.123

C, 52.07; H, 5.62; Na, 9.49; O, 26.42; P, 6.39

125-02-0

Prednisolone;50-24-8;Prednisolone hemisuccinate;2920-86-7;Prednisolone acetate;52-21-1; 630-67-1 (sodium metazoate); 72064-79-0 (valerate acetate); 125-02-0 (Na+ phosphate); 50-24-8 (free)

441409

White to light yellow solid powder

100 °C

361.9ºC

4.17E-21mmHg at 25°C

2.551

2

8

3

32

878

7

C[C@@]1([C@@]2(O)C(COP([O-])([O-])=O)=O)[C@](CC2)([H])[C@@](CCC3=CC4=O)([H])[C@]([C@]3(C=C4)C)([H])[C@@H](O)C1.[Na+].[Na+]

VJZLQIPZNBPASX-OJJGEMKLSA-L

InChI=1S/C21H29O8P.2Na/c1-19-7-5-13(22)9-12(19)3-4-14-15-6-8-21(25,17(24)11-29-30(26,27)28)20(15,2)10-16(23)18(14)19;;/h5,7,9,14-16,18,23,25H,3-4,6,8,10-11H2,1-2H3,(H2,26,27,28);;/q;2*+1/p-2/t14-,15-,16-,18+,19-,20-,21-;;/m0../s1

disodium;[2-[(8S,9S,10R,11S,13S,14S,17R)-11,17-dihydroxy-10,13-dimethyl-3-oxo-7,8,9,11,12,14,15,16-octahydro-6H-cyclopenta[a]phenanthren-17-yl]-2-oxoethyl] phosphate

prednisolone; Prednisolone Phosphate Sodium; prednisolone sodium phosphate; 125-02-0; Prednisolone phosphate sodium; Phortisolone; Metreton; Orapred; Predair; Pediapred; AKOS016010152; AK115681; sodium phosphate Predsol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O: ~50 mg/mL (~103.2 mM)

Solubility in Formulation 1: 100 mg/mL (206.45 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)