| 生物活性 |
Fludrocortisone (9α-Fludrocortisone) is an orally active mineralocorticoid and glucocorticoid receptor agonist. Fludrocortisone suppresses pro-inflammatory cytokine expression, reduces CCL2, IL-6, IL-8 levels, upregulates mineralocorticoid receptor (MR) expression, induces PI3K/Akt, GSK-3β, CREB, ERK1/2, mTOR phosphorylation, blocks Tau hyperphosphorylation, prevents apoptosis, promotes survival and proliferation, enhances renal sodium and water transport, increases plasma volume and blood pressure, reduces plasma potassium and renin activity, stimulates erythropoietin expression, modulates uterine receptivity genes, and reverses PP242-induced MUC1 upregulation. Fludrocortisone can be used for the research of congenital adrenal hyperplasia, postural hypotension, and adrenal insufficiency[1][2].
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| 体外研究 |
Fludrocortisone (0.01-10 μg/μ L; 12 h) showed no cytotoxicity towards MIO-M1, ARPE19, and 661W human retinal cell lines at concentrations not exceeding 1 μ g/μ L. Only a decrease in cell viability was observed in MIO-M1 and 661W cells at concentrations of 10 μg/μL [1].
Fludrocortisone (1 μg/μL; 12 h) can effectively inhibit the mRNA expression of CCL2, IL-6, and IL-8 in the human M ü ller glial cell line MIO-M1 induced by IL-1 β and TNF - α [1].
Fludrocortisone (1 μM; 24 h) can upregulate the expression of mineralocorticoid receptor (MR) genes in AHPs without altering glucocorticoid receptor (GR) expression [2].
Fludrocortisone (0.5-2 μM; 24 h) can promote cell survival and proliferation, and reduce cell apoptosis in growth factor deficient AHP, with 1 μM being the optimal concentration [2].
Fludrocortisone (1 μM) can activate the cAMP/PKA/CREB signaling pathway in AHPs, which is necessary for its pro survival and pro proliferative effects [2].
Fludrocortisone (1 μM) can activate the PI3K/Akt and mTOR/p70S6K signaling pathways, and inactivate GSK-3 β, which is necessary for its protective effect in growth factor deficient AHP [2].
Fludrocortisone (1 μM) can protect AHPs from toxic damage induced by A β 1-42 by promoting survival and proliferation, reducing cell apoptosis, restoring Akt/GSK-3 β phosphorylation, and blocking Tau hyperphosphorylation.
Fludrocortisone (4-6 h) can stimulate the mRNA expression of Epo, HIF2 α, HIF1 α, and PHD2 in microscopically isolated rat renal unit segments, mouse renal unit segments, and peritubular cells, which is significantly increased compared to the baseline state.
Fludrocortisone (6 h, 72 h) significantly upregulated the expression of Epo protein in rat renal cortex compared to baseline, but had no effect on the expression of Epo protein in rat liver.
Fludrocortisone (4-6 h) can stimulate the production of Epo protein in mouse MAL, CCD, and OMCD, and is specifically upregulated in collecting duct type A leap cells. Its effect decreases to baseline levels at 72 h.
Cell Viability Assay
| Cell Line: |
human MIO-M1 Müller glial cells, ARPE19 retinal pigment epithelial cells, 661W cone photoreceptor-like cells |
| Concentration: |
0.01-10 μg/μL |
| Incubation Time: |
12 hours |
| Result: |
| Did not significantly alter cell viability in ARPE19 cells at all tested concentrations. Showed unchanged viability in MIO-M1 and 661W cells at 0.01, 0.1, and 1 μg/μL; caused a slight but statistically significant reduction in viability only at the highest concentration of 10 μg/μL in MIO-M1 and 661W cells. |
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Real Time qPCR
| Cell Line: |
human MIO-M1 Müller glial cells |
| Concentration: |
1 μg/μL |
| Incubation Time: |
12 hours (co-incubated with IL-1β stimulation) |
| Result: |
Completely abrogated IL-1β-induced significant increases in CCL2, IL-6, and IL-8 mRNA expression, suppressing expression to near baseline levels. |
Real Time qPCR
| Cell Line: |
human MIO-M1 Müller glial cells |
| Concentration: |
1 μg/μL |
| Incubation Time: |
12 hours (co-incubated with TNF-α stimulation) |
| Result: |
Sharply reduced TNF-α-induced significant increases in CCL2, IL-6, and IL-8 mRNA expression, suppressing expression to near baseline levels. |
Real Time qPCR
| Cell Line: |
human MIO-M1 Müller glial cells |
| Concentration: |
1 μg/μL |
| Incubation Time: |
12 hours (co-incubated with IL-1β stimulation) |
| Result: |
Had its suppressive effect on IL-1β-induced CCL2, IL-6, and IL-8 expression completely abrogated by co-incubation with glucocorticoid receptor antagonist RU486. |
Real Time qPCR
| Cell Line: |
Adult rat hippocampal progenitor cells (AHPs) |
| Concentration: |
1 μM |
| Incubation Time: |
24 hours |
| Result: |
Strongly increased mineralocorticoid receptor (MR) gene expression; showed no significant change in glucocorticoid receptor (GR) gene expression. |
Cell Viability Assay
| Cell Line: |
Adult rat hippocampal progenitor cells (AHPs) |
| Concentration: |
0.01, 0.1, 0.5, 1, 2 μM |
| Incubation Time: |
24 hours |
| Result: |
Progressively increased cell survival and proliferation at 0.5, 1, and 2 μM; strongly reduced serum starvation-induced apoptosis (via decreased caspase-3 activity) at 0.5, 1, and 2 μM; identified 1 μM as the optimal concentration for subsequent experiments. |
Western Blot Analysis
| Cell Line: |
Adult rat hippocampal progenitor cells (AHPs) |
| Concentration: |
1 μM (cAMP/CREB phosphorylation assays); 1 μM (functional validation with KT5720) |
| Incubation Time: |
5, 15, 30, 60, 90 min (cAMP/CREB phosphorylation assays); 24 h (functional validation with KT5720) |
| Result: |
Increased intracellular cAMP levels, with a peak at 15 min and sustained elevation through 90 min; promoted CREB phosphorylation (Ser133), peaking at 15-30 min and returning to near basal levels by 90 min; had induced survival and proliferation reduced by co-treatment with PKA inhibitor KT5720. |
Western Blot Analysis
| Cell Line: |
adult rat hippocampal progenitor cells (AHPs) |
| Concentration: |
1 μM (phosphorylation assays); 1 μM (functional validation with wortmannin/rapamycin) |
| Incubation Time: |
5, 15, 30, 60, 90 min (phosphorylation assays); 24 h (functional validation with wortmannin/rapamycin) |
| Result: |
Induced time-dependent phosphorylation of Akt (peaking at 30 min, sustained through 90 min), GSK-3β (sustained elevation from 5 to 90 min), and p70S6K (progressive elevation through 90 min); had induced survival, proliferation, and antiapoptotic effects reduced by co-treatment with Wortmannin or Rapamycin. |
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| 体内研究 |
Fludrocortisone (1 μg/μL; intravitreal injection (3 μL); Single dose plays a neuroprotective role in a mouse model of photooxidative retinal degeneration, maintaining retinal structure and function, reducing apoptosis of photoreceptor cells, and inhibiting macrophage infiltration [1].
Fludrocortisone (1.5 mg/kg; i.p.; twice on Days 4 and 5 of pregnancy) can regulate uterine receptivity in pregnant BALB/c mice by upregulating the expression of LIF, HB-EGF, Msx.1, and miRNA Let-7a and activating the ERK1/2-mTOR pathway without altering the expression of MUC1 or miRNA 223-3p.
The combination administration of Fludrocortisone (1.5 mg/kg; i.p.; twice on Days 4 and 5 of pregnancy) and PP242 (30 mg/kg) can reverse the upregulation of MUC1 induced by PP242 and restore the upregulation of key uterine receptivity genes (LIF, HB-EGF, Msx.1, miRNA Let-7a) in pregnant BALB/c mice.
Fludrocortisone (2.5 mg/100 g BW/day; i.p.; daily) can stimulate renal Epo mRNA expression (peak at 4 hours) and Epo protein expression (6 hours and 72 hours) in male Sprague Dawley rats. A statistically significant increase in plasma Epo concentration was observed 6 hours after administration.
| Animal Model: |
C57BL/6J (60 post-natal days old) |
| Dosage: |
1 μg/μL |
| Administration: |
intravitreal injection; single dose |
| Result: |
Preserved outer nuclear layer thickness at locations 1-2 mm and 2-3 mm superior to the optic nerve compared to controls; Achieved significantly higher a-wave and b-wave ERG responses; Reduced the number of TUNEL+ apoptotic photoreceptors significantly compared to vehicle-treated mice; Decreased the number of subretinal IBA1+ macrophages significantly compared to vehicle-treated mice. |
| Animal Model: |
BALB/c (8-week-old virgin female; pregnancy model, estrous induced, mated with fertile males, vaginal plug confirmed) |
| Dosage: |
1.5 mg/kg |
| Administration: |
i.p.; twice on Days 4 and 5 of pregnancy at 8:00 a.m. |
| Result: |
Did not significantly alter mRNA expression of mucin-1 (MUC1) or miRNA 223-3p; significantly upregulated mRNA expression of leukemia inhibitory factor (LIF), heparin-binding epidermal growth factor (HB-EGF, Msx.1, and miRNA Let-7a; significantly increased phosphorylated ERK1/2 (p-ERK1/2) and phosphorylated mammalian target of rapamycin (p-mTOR) levels; did not change phosphorylated eukaryotic translation initiation factor 4E-binding protein 1 (p-4E-BP1) levels significantly; induced round epithelial cell nuclei, cuboidal endometrial epithelium, no detectable glycoprotein layer on the epithelial surface, a thin continuous basement membrane beneath epithelial cells, and proliferating fibroblast-like stromal cells. |
| Animal Model: |
BALB/c (8-week-old virgin female; pregnancy model, estrous induced, mated with fertile males, vaginal plug confirmed) |
| Dosage: |
1.5 mg/kg (fludrocortisone); 30 mg/kg (PP242) |
| Administration: |
i.p.; twice on Days 4 and 5 of pregnancy at 8:00 a.m. (fludrocortisone); co-administered with PP242 |
| Result: |
Significantly upregulated mRNA expression of LIF, HB-EGF, Msx.1, and miRNA Let-7a compared to PP242-only group; significantly downregulated mRNA expression of MUC1 compared to PP242-only group; did not change miRNA 223-3p expression significantly compared to PP242-only group. |
| Animal Model: |
Sprague Dawley (male) |
| Dosage: |
2.5 mg/100 g BW/day |
| Administration: |
i.p.; daily |
| Result: |
Increased Epo, HIF2α, and HIF1α mRNA expression in renal cortex at 4 hours post-injection; the stimulating effect disappeared by 72 hours. Increased Epo protein expression in renal cortex at 6 and 72 hours. Increased plasma Epo concentration at 6 hours (statistically significant elevation vs. vehicle), but not at 72 hours. Did not alter Epo mRNA or protein expression in the liver. |
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| 分子式 |
C21H29FO5
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| 分子量 |
380.4503632
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| CAS号 |
127-31-1
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| 中文名称 |
氟氢可的松
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| 运输条件 |
Room temperature in continental US; may vary elsewhere.
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| 储存方式 |
| Powder |
-20°C |
3 years |
| |
4°C |
2 years |
| In solvent |
-80°C |
6 months |
| |
-20°C |
1 month |
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| 溶解性数据 |
In Vitro:
DMSO : 100 mg/mL (262.85 mM; Need ultrasonic)
配制储备液
| 浓度 溶剂体积 质量 |
1 mg |
5 mg |
10 mg |
| 1 mM |
2.6285 mL |
13.1423 mL |
26.2847 mL |
| 5 mM |
0.5257 mL |
2.6285 mL |
5.2569 mL |
| 10 mM |
0.2628 mL |
1.3142 mL |
2.6285 mL |
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。
In Vivo:
请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：
——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用；以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶
-
1.
请依序添加每种溶剂： 10% DMSO → 90% (20% SBE-β-CD in saline)
Solubility: ≥ 2.5 mg/mL (6.57 mM); Clear solution
此方案可获得 ≥ 2.5 mg/mL (6.57 mM，饱和度未知) 的澄清溶液。
以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。
-
2.
请依序添加每种溶剂：10% DMSO → 90% corn oil
Solubility: ≥ 2.5 mg/mL (6.57 mM); Clear solution
此方案可获得 ≥ 2.5 mg/mL (6.57 mM，饱和度未知) 的澄清溶液，此⽅案不适⽤于实验周期在半个⽉以上的实验。
以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL ⽟⽶油中，混合均匀。
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Data Sheet