| Size | Price | Stock | Qty |
|---|---|---|---|
| 250mg |
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| 500mg |
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| 1g |
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Purity: ≥95%
| Targets |
Natural product from beets; acetylcholinesterase; anticancer
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|---|---|
| ln Vitro |
Dysfunction of the cholinergic system and increased oxidative stress have a crucial role in cognitive disorders including Alzheimer's disease (AD). Here, we have investigated the protective effects of betanin, a novel acetylcholinesterase (AChE) inhibitor, on hydrogen peroxide (H2O2)-induced cell death in PC12 cells.[1]
Methods and results: The protective effects were assessed by measuring cell viability, the amount of reactive oxygen species (ROS) production, AChE activity, cell damage, and apoptosis using resazurin, 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA), Ellman method, lactate dehydrogenase (LDH) release, propidium iodide (PI) staining and flow cytometry, and Western blot analysis. H2O2 (150 µM) resulted in cell viability reduction and apoptosis induction while, pretreatment with the betanin (10, 20, and 50 μM) and N-Acetyl-L-cysteine (NAC) (2.5 and 5 mM) significantly increased the viability (P < 0.05, P < 0.01 and P < 0.001) and at 5-50 μM betanin decreased ROS amount (P < 0.05, P < 0.01 and P < 0.001). Whereas, pretreatment with the betanin (10, 20, and 50 μM) decreased AChE activity (P < 0.001), also at 20 and 50 μM betanin reduced the release of LDH (P < 0.001), and at 10-50 μM decreased the percentage of apoptotic cells (P < 0.001). Apoptosis biomarkers such as cleaved poly (ADP-ribose) polymerase (PARP) (P < 0.01 and P < 0.001) and cytochrome c (P < 0.05 and P < 0.001) were attenuated after pretreatment of PC12 cells with betanin at 10-20 μM and 10-50 μM respectively. Indeed, survivin (P < 0.001) increased after pretreatment of cells with betanin at 10-20 μM.[1] Conclusions: Overall, betanin may use the potential to delay or prevent cell death caused by AD through decreasing the activity of AChE as well as attenuating the expression of proteins involved in the apoptosis pathway.[1] |
| ln Vivo |
The heat exposure and white noise can induce damage on reproductive organs. The main objective of this study is to observe, if betanin administration could ameliorate oxidative stress, apoptosis and inflammation in testis of rodents following noise and scrotal hyperthermia exposure. Wistar rats were divided into 6 groups; control, betanin, noise, hyperthermia and two treatment groups. Scrotal hyperthermia model was performed by heat exposure of rat testicular (43 °C) for 15 min and 3 times per weeks for 14 days. Noise induction model was done following exposure of rats with 100-dB noise level for 14 days and 8 h daily similar to real exposure condition in human. Betanin was administrated at the sub-effective dose (15 mg/kg) by gavage route for 4 weeks (5 times a week) to male rats. The animals were euthanized and testis were dissected and stored at -80 °C. Then, the oxidative stress biomarkers (MDA and GSH), apoptosis (cytochrome c & Annexin V), and inflammatory cytokines (TNF-α & IL-6) were measured by the real time polymerase chain reaction (RT-PCR) of testis collected samples. The data output demonstrates the impact of noise and hyperthermia in testicular toxicity induction by mitigating oxidative damage, apoptosis and inflammatory mediators. Following treatment with 15 mg/kg per day of betanin, lipid peroxidation and GSH content have been modulated, and TNF-α and IL-6 gene expression has been declined. Our results revealed that in Wistar rats, betanin displays protective effects against noise and scrotal hyperthermia-induced acute testicular toxicity through the inhibition of oxidative stress, apoptosis, and inflammation[2].
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| Enzyme Assay |
AChE inhibitory activity assay[1]
For this purpose, 1.0 × 106 PC12 cells were grown in a T25 flask. To extract the enzyme, the supernatant of the cells was discarded and the cells were trypsinized. Then the supernatant discarded and, the cell plate homogenized in: 10 mM Tris (pH 7.2), 1 M sodium chloride (NaCl), 50 mM magnesium chloride (MgCl2) and triton X-100 (1%). The resulting suspension was centrifuged at 15,000 rpm for 10 min and the supernatant used as an enzyme source. Then, the AChE inhibitory activity of betanin was measured by the Ellman method with some modifications. The amount of 2 mL of phosphate buffer with pH 7.6, 60 μL of 5-dithiobis-2-nitrobenzoic acid (DTNB) reagent, 30 μL of different concentrations of betanin (10, 20 and 50 μM) and 20 μL of extracted enzyme were added to 24-well plates for 15 min. Then, 20 μL of acetylthiocholine iodide (100 mM) was added as enzyme substrate. Finally, the absorbance was measured by an ELISA reader at 412 nm. Measurement of LDH release[1] 1.0 × 105 PC12 cells were seeded 12-well plates and were pretreated with betanin (20 and 50 μM) and then incubated for 24 h before exposure to H2O2 (150 µM). Finally, we collected supernatant and read the absorbance at 490 nm by ELISA plate reader. |
| Cell Assay |
Cell viability analysis[1]
First, we evaluated betanin cytotoxicity by resazurin assay. For this purpose, PC12 cells (1.0 × 104) were seeded in 96-well plates. Betanin was dissolved in DMSO to obtain a 50 mM stock solution.[1] After 24 h, the cells were treated with different concentrations of betanin (5–50 μM) and NAC (1.25, 2.5, 5 and 10 mM). After an additional 24 h, the resazurin reagent (20 µl; 10 mg/mL) was added to each well and maintained at 37 °C and 5% CO2 for 4–6 h. The absorbance of the samples was read at 570 and 600 nm with a microplate reader. Afterward based on results, we assessed the effects of different concentrations of betanin (5–50 μM) and NAC (1.25, 2.5 and 5 mM) on H2O2 induced cytotoxicity on PC12 cells according to previously described method. Intracellular ROS production[1] To indicate intracellular ROS production, 1.0 × 104 PC12 cells were seeded in 96-well plates. Cells were pretreated with betanin (5–50 μM) and NAC (1.25, 2.5 and 5 mM) for 24 h before exposed to H2O2 (150 µM). After 4 h, we added 2.5 μM DCFH-DA (Stock solution: 40 mM in DMSO) reagent to each well for 30 min. Finally, we assessed the fluorescent emission at 525 nm after excitation at 488 nm using a microplate reader to measure and compare ROS generation to the related control. |
| Animal Protocol |
48 rats were divided into 6 groups without specific pattern: (1) Control group (n = 6), received only vehicle (saline) via orally administration based on scrotal hyperthermia (n = 3) or noise induction models (n = 3); (2) betanin group (n = 6) and received 15 mg kg−1 betanin dissolved in saline orally for 30 days every other day based on scrotal hyperthermia (n = 3) or noise induction models (n = 3). The pilot study revealed that non-significant difference between control or/and betanin treated rats in normal condition. Therefore, the results of two different control or/and betanin groups were used as one similar group in two different animal models. The dosage and duration of betanin administration were selected based on pilot and other previous studies in antioxidant enzyme activity test in noise or scrotal hyperthermia rat models. Our data showed the similar response in antioxidant enzyme activity in betanin treated between 15 and 20 mg/kg in noise or scrotal hyperthermia rats and the lowest dose was selected in present study [23,29]. Betanin in 15 mg/kg per day was administered by intra-gastric gavage method (i.g., saline as vehicle) for five consecutive weeks following; (3) Animals exposure with scrotal hyperthermia induction model (H, n = 6), every other day for five consecutive weeks; (4) Animals exposure with noise (100 dB) 8h per day for 14 days (N, n = 6); (5) Treated of scrotal hyperthermia model animals with betanin (H + B, n = 6); (6) Treated of noise model animals with betanin (H + B, n = 6).[2]
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| References |
[1]. Protective effects of betanin, a novel acetylcholinesterase inhibitor, against H2O2-induced apoptosis in PC12 cells. Mol Biol Rep . 2024 Sep 16;51(1):986.
[2]. Protective impact of Betanin against noise and scrotal hyperthermia on testicular toxicity in Wistar rat: Role of apoptosis, oxidative stress and inflammation. Heliyon . 2024 Sep 24;10(19):e38289. |
| Additional Infomation |
Colorectal cancer (CRC) is a common and life-threatening neoplastic disease that continues to pose a formidable challenge to global health. The present work was performed to evaluate the anticancer properties of betanin and betanin (BT) loaded starch nanoparticles (S-BT). The BT and S-BT were characterized by DLS, SEM, UV spectroscopy, XPS and FTIR. The cytotoxic effect was assessed by MTT and LDH assay. The apoptotic potential of BT and S-BT was assessed by DCFDA, Rh123, AO/EB and DAPI staining methods. Cell cycle arrest was depicted using flow cytometry. The antimetastatic potential of BT and S-BT was evaluated by wound healing assay. The S-BT showed a spherical morphology with a size of 175 nm. The betanin contained SNPs were found to have strong encapsulation efficiency and favorable release profiles. Both BT and S-BT exhibited cytotoxicity in SW480 cells but S-BT displayed increased cytotoxicity when compared to BT alone. Loss of mitochondrial membrane potential, nuclear fragmentation, chromatin condensation and generation of ROS, all indicative of apoptotic mode of cell death, were revealed by fluorescence imaging. The cells were arrested in the G2M phase. Moreover, both BT and S-BT were able to inhibit the migratory potential of SW480 cells. Overall, our results indicated that both BT and S-BT were able to induce anticancer effects; and, S-BT was found to have increased therapeutic efficacy when compared to BT alone. https://pubmed.ncbi.nlm.nih.gov/39297057/
In this study, H2O2 induces cell apoptosis through main apoptosis biomarkers PARP, cytochrome c and, survivin as the possible mechanism of apoptosis for in vitro model of AD. Pretreatment with betanin reduced cell injury and protected PC12 cells with antioxidant and anti-apoptotic properties. Also in the present study, for the first time, the inhibitory effects of betanin on AChE activity were confirmed. Our analysis suggests that betanin has the potential to be considered as neuroprotective agents against neural death.[1] |
| Molecular Formula |
C24H26N2O13
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|---|---|
| Molecular Weight |
550.47
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| Exact Mass |
550.143
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| Elemental Analysis |
C, 52.37; H, 4.76; N, 5.09; O, 37.78
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| CAS # |
7659-95-2
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.8±0.1 g/cm3
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| Boiling Point |
983.5±75.0 °C at 760 mmHg
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| Flash Point |
548.6±37.1 °C
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| Vapour Pressure |
0.0±0.3 mmHg at 25°C
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| Index of Refraction |
1.740
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| LogP |
-4.49
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| SMILES |
C(N1[C@@H](CC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=C(O)C=C12)C(=O)O)=CC1=CC(C(=O)O)=N[C@H](C(=O)O)C1
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| InChi Key |
DHHFDKNIEVKVKS-UWTTYFQQSA-N
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| InChi Code |
InChI=1S/C24H26N2O13/c27-8-17-18(29)19(30)20(31)24(39-17)38-16-6-10-5-14(23(36)37)26(13(10)7-15(16)28)2-1-9-3-11(21(32)33)25-12(4-9)22(34)35/h1-3,6-7,12,14,17-20,24,27,29-31H,4-5,8H2,(H4,28,32,33,34,35,36,37)/t12?,14?,17-,18-,19+,20-,24-/m1/s1
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| Chemical Name |
(E)-1-((Z)-2-(2,6-dicarboxy-2,3-dihydropyridin-4(1H)-ylidene)ethylidene)-6-hydroxy-5-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)indolin-1-ium-2-carboxylate
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| Synonyms |
7659-95-2; 1-[2-(2,6-dicarboxy-2,3-dihydro-1H-pyridin-4-ylidene)ethylidene]-6-hydroxy-5-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-2,3-dihydroindol-1-ium-2-carboxylate; 1-[(2E)-2-(2,6-Dicarboxy-2,3-dihydro-1H-pyridin-4-ylidene)ethylidene]-6-hydroxy-5-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-2,3-dihydroindol-1-ium-2-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 |
| 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 | 1.8166 mL | 9.0831 mL | 18.1663 mL | |
| 5 mM | 0.3633 mL | 1.8166 mL | 3.6333 mL | |
| 10 mM | 0.1817 mL | 0.9083 mL | 1.8166 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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