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| ln Vitro |
In LPS-stimulated RAW 264.7 macrophages, 4′-Hydroxywogonin (8-Methoxyapigenin; 0.5-15 μM; 0-24 h) suppresses iNOS and COX-2 expression, which prevents NO and PGE2 production[1]. It also displays low cytotoxicity. -Induction of pro-inflammatory cytokines by LPS in RAW 264.7 macrophages and inhibition of LPS-induced activation of NF-κB are both inhibited by hydroxywogonin (0.5-15 μM; 1 and 12 h)[1]. RAW 264.7 macrophages treated with 4′-hydroxywogonin (0.5–15 μM; 1 h) are less susceptible to LPS-induced IκB–α degradation, TAK and IKK activation, and MAPK and AKTin phosphorylation[1]. In LPS-stimulated RAW 264.7 macrophages, 4′-hydroxywogonin (0.5–15 μM; 24 h) minimizes ROS production[1]. When exposed to concentrations and times that vary, 4′-hydroxywogonin (0–10 μg/mL; 24 h) diminishes the survival of SW620 cells and lowers the expression of VEGF-A (vascular endothelial growth factor–A), the main pro-angiogenic cytokine in tumor angiogenesis, both in mRNA and protein[2]. Apoptosis is induced and the expression of C-MYC, BCL-2, and cleaved caspase 3 is reduced by 4′-hydroxywogonin (24 h; SUP-B15 and Jurkat cells)[3].
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| ln Vivo |
In a mouse model, 4′-hydroxywogonin (10 and 20 mg/kg; ip; male C57BL/6 mice) reduces LPS-induced acute lung damage (ALI)[1].
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: RAW 264.7 macrophages Tested Concentrations: 0.5, 5 and 15 μM Incubation Duration: 24 hrs (hours) Experimental Results: Had low cytotoxicity in RAW 264.7 macrophages. Western Blot Analysis[1] Cell Types: RAW 264.7 macrophages Tested Concentrations: 0.5, 5 and 15 μM Incubation Duration: 1 hrs (hours) Experimental Results: Attenuated the increase of iNOS and COX-2 mRNA expression induced by LPS in RAW 264.7 cells. Western Blot Analysis[1] Cell Types: RAW 264.7 macrophages Tested Concentrations: 0.5, 5 and 15 μM Incubation Duration: 1 and 12 hrs (hours) Experimental Results: decreased TNF-α, IL-6 and IL-1β mRNA expression in a dose-dependent manner. Inhibited LPS-induced p65 phosphorylation and nuclear translocation. Western Blot Analysis[1] Cell Types: RAW 264.7 macrophages Tested Concentrations: 0.5, 5 and 15 μM Incubation Duration: 1 hrs (hours) Experimental Results: Attenuated LPS induced IκB-α degradation. Attenuated the phosphorylation of ERK1/2 and p38 in a dose-dependent manner. decreased the intensity of the TAK1/TAB1 band. Western Blot Analysis[2] Cell Types: SW620 cells Concen |
| Animal Protocol |
Animal/Disease Models: Male C57BL/6 mice (6-8 weeks old; 20 g) with acute lung injury model[1]
Doses: 10 and 20 mg/kg Route of Administration: intraperitoneal (ip)injection, 12 and 1 h before LPS treatment Experimental Results: Had potential protective effects against inflammation in LPS induced ALI mice. Attenuated the degree of leukocyte infiltration. |
| References |
[1]. Fan C, et, al. 4'-Hydroxywogonin suppresses lipopolysaccharide-induced inflammatory responses in RAW 264.7 macrophages and acute lung injury mice. PLoS One. 2017 Aug 8;12(8):e0181191.
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| Molecular Formula |
C16H12O6
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|---|---|
| Molecular Weight |
300.26
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| CAS # |
57096-02-3
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| SMILES |
O1C(=C([H])C(C2C(=C([H])C(=C(C1=2)OC([H])([H])[H])O[H])O[H])=O)C1C([H])=C([H])C(=C([H])C=1[H])O[H]
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3304 mL | 16.6522 mL | 33.3045 mL | |
| 5 mM | 0.6661 mL | 3.3304 mL | 6.6609 mL | |
| 10 mM | 0.3330 mL | 1.6652 mL | 3.3304 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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