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In H9c2 rat cardiomyocytes injured by H₂O₂ (0.5 mM), Hederagenin 28-O-beta-D-glucopyranosyl ester (10 μM, 20 μM, 40 μM) exerted dose-dependent protective effects.
1. Cell viability: MTT assay showed that H₂O₂ alone reduced cell viability to 45.2% of the control group; pretreatment with 10 μM, 20 μM, and 40 μM Hederagenin 28-O-beta-D-glucopyranosyl ester increased viability to 58.7%, 71.3%, and 82.3% respectively [1] 2. Oxidative stress markers: DCFH-DA staining revealed that 40 μM Hederagenin 28-O-beta-D-glucopyranosyl ester reduced intracellular reactive oxygen species (ROS) levels by 58.6% compared to the H₂O₂ group. It also decreased malondialdehyde (MDA, a lipid peroxidation product) content by 42.1% (40 μM dose) and increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) by 35.8% and 41.5% respectively (40 μM dose) [1] |
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| Cell Assay |
1. Cell culture and grouping: H9c2 rat cardiomyocytes were cultured in DMEM medium containing 10% fetal bovine serum at 37°C with 5% CO₂. Cells were divided into 4 groups: control group (no treatment), H₂O₂ group (0.5 mM H₂O₂ for 4 hours), and two Hederagenin 28-O-beta-D-glucopyranosyl ester pretreatment groups (10 μM, 20 μM, 40 μM drug for 2 hours before H₂O₂ exposure) [1]
2. Cell viability detection: After treatment, cells were incubated with MTT reagent (5 mg/mL) for 4 hours at 37°C. The supernatant was removed, and DMSO was added to dissolve formazan crystals. Absorbance was measured at 570 nm to calculate cell viability [1] 3. Oxidative stress detection: For ROS measurement, cells were loaded with DCFH-DA probe (10 μM) for 30 minutes, then washed and observed under a fluorescence microscope to quantify fluorescence intensity. For MDA, SOD, and GSH-Px detection: cell homogenates were prepared, and commercial assay kits were used to measure their levels/activities according to the kit instructions [1] |
| Toxicity/Toxicokinetics |
In normal H9c2 cardiomyocytes (untreated with H₂O₂), a concentration of up to 40 μM of Hederagenin 28-O-β-D-glucopyranose ester did not show significant cytotoxicity. MTT assay results showed that 24 hours after drug treatment, cell viability remained above 95% compared with the control group, indicating that there was no significant damage to normal cardiomyocytes [1].
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| References | |
| Additional Infomation |
Ivy saponin 28-O-β-D-glucopyranoside is a triterpenoid saponin, which is a carboxylic acid ester formed by the condensation of the carboxyl group of ivy saponin with β-D-glucopyranoside. It has been isolated from walnut (Juglans sinensis). Ivy saponin has plant metabolites and anti-inflammatory activities. It is a triterpenoid saponin, a pentacyclic triterpenoid compound, a monosaccharide derivative, a β-D-glucopyranoside and a carboxylic acid ester. Its function is related to ivy saponin. It is derived from the hydride of oleanane.
It has been reported that ivy saponin 28-O-β-D-glucopyranoside is found in Achyranthes bidentata, Acer rubrum, and other organisms with relevant data. Ivy saponin 28-O-β-D-glucopyranoside is a novel triterpenoid saponin isolated from the leaves of the traditional Chinese medicine plant Ilex chinensis[1]. Ivy saponin 28-O-β-D-glucopyranose The protective effect of H₂O₂-induced cardiomyocyte damage is mainly achieved by inhibiting oxidative stress: it can reduce the generation of reactive oxygen species (ROS) and lipid peroxidation (reduce malondialdehyde (MDA) level), while enhancing the activity of endogenous antioxidant enzymes (superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px)), thereby restoring the redox balance of cells [1]. |
| Molecular Formula |
C36H58O9
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|---|---|
| Molecular Weight |
634.8403
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| Exact Mass |
634.408
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| CAS # |
53931-25-2
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| PubChem CID |
21120798
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| Appearance |
White to off-white solid powder
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| Density |
1.27±0.1 g/cm3
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| Boiling Point |
729.7±60.0 °C at 760 mmHg
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| Melting Point |
197-202 ºC (methanol )
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| Flash Point |
220.2±26.4 °C
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| Vapour Pressure |
0.0±5.4 mmHg at 25°C
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| Index of Refraction |
1.596
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| LogP |
6.68
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| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
45
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| Complexity |
1200
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| Defined Atom Stereocenter Count |
14
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| SMILES |
O([H])C1([H])C([H])([H])C([H])([H])[C@@]2(C([H])([H])[H])[C@]([H])([C@]1(C([H])([H])[H])C([H])([H])O[H])C([H])([H])C([H])([H])[C@@]1(C([H])([H])[H])[C@]3(C([H])([H])[H])C([H])([H])C([H])([H])[C@@]4(C(=O)OC5([H])C([H])(C([H])(C([H])(C([H])(C([H])([H])O[H])O5)O[H])O[H])O[H])C([H])([H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[C@]4([H])C3=C([H])C([H])([H])[C@@]12[H]
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| InChi Key |
WJMMBVSOQPALFO-DLQTVUGOSA-N
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| InChi Code |
InChI=1S/C36H58O9/c1-31(2)13-15-36(30(43)45-29-28(42)27(41)26(40)22(18-37)44-29)16-14-34(5)20(21(36)17-31)7-8-24-32(3)11-10-25(39)33(4,19-38)23(32)9-12-35(24,34)6/h7,21-29,37-42H,8-19H2,1-6H3/t21-,22+,23+,24+,25-,26+,27-,28+,29-,32-,33-,34+,35+,36-/m0/s1
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| Chemical Name |
[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] (4aS,6aR,6aS,6bR,8aR,9R,10S,12aR,14bS)-10-hydroxy-9-(hydroxymethyl)-2,2,6a,6b,9,12a-hexamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylate
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| HS Tariff Code |
2934.99.9001
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| Storage |
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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5752 mL | 7.8760 mL | 15.7520 mL | |
| 5 mM | 0.3150 mL | 1.5752 mL | 3.1504 mL | |
| 10 mM | 0.1575 mL | 0.7876 mL | 1.5752 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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