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| Targets |
The action target of 8-Hydroxyguanosine is B lymphocytes (mediating immunostimulating activity) [3]
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| ln Vitro |
Immunostimulating activity on B lymphocytes: Murine splenic B lymphocytes were isolated and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum. The cells were treated with 8-Hydroxyguanosine at concentrations of 10 μg/mL, 20 μg/mL, 40 μg/mL, and 80 μg/mL for 72 hours. [3H]-thymidine incorporation assay showed 8-Hydroxyguanosine dose-dependently promoted B lymphocyte proliferation: 40 μg/mL 8-Hydroxyguanosine increased the proliferation rate by 2.3-fold compared to the control group, and 80 μg/mL increased it by 3.1-fold. Additionally, enzyme-linked immunosorbent assay (ELISA) revealed that 8-Hydroxyguanosine enhanced the secretion of IgG and IgM by B lymphocytes: 80 μg/mL 8-Hydroxyguanosine increased IgG and IgM levels by 1.8-fold and 1.6-fold, respectively [3]
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| ln Vivo |
1. As a marker of oxidative DNA/RNA damage in rat liver: Male Sprague-Dawley rats were divided into control group and 2-nitropropane-treated group. The treated group received 2-nitropropane (100 mg/kg body weight) via oral gavage twice daily for 14 days. At the end of treatment, liver tissues were collected, and DNA/RNA was extracted. High-performance liquid chromatography (HPLC) analysis showed that the level of 8-Hydroxyguanosine in liver DNA of the treated group was 4.2-fold higher than that of the control group, and the level in liver RNA was 3.8-fold higher. This indicated that 2-nitropropane-induced oxidative stress caused DNA and RNA damage, with 8-Hydroxyguanosine serving as a key biomarker [1]
2. Concentration change in cerebrospinal fluid (CSF) of Alzheimer's disease (AD) patients: CSF samples were collected from 15 AD patients and 15 age-matched healthy controls. The concentration of 8-Hydroxyguanosine in CSF was detected by immunodot assay. The results showed that the concentration of 8-Hydroxyguanosine in AD patients was 2.7-fold higher than that in healthy controls (AD: 18.6 ± 3.2 pg/mL vs. control: 6.9 ± 1.5 pg/mL), suggesting that 8-Hydroxyguanosine could be a biomarker for oxidative stress in AD [2] |
| Enzyme Assay |
1. Detection of 8-Hydroxyguanosine in rat liver DNA/RNA (HPLC method): Liver tissues were homogenized and treated with proteinase K to remove proteins. DNA and RNA were isolated using phenol-chloroform extraction and ethanol precipitation. The samples were hydrolyzed with nuclease P1 and alkaline phosphatase to convert nucleic acids into nucleosides. The hydrolyzed samples were filtered through a 0.22 μm membrane and analyzed by HPLC equipped with a UV detector (detection wavelength: 254 nm). The concentration of 8-Hydroxyguanosine was calculated using a standard curve prepared with authentic 8-Hydroxyguanosine [1]
2. Detection of 8-Hydroxyguanosine in human CSF (immunodot assay): CSF samples were spotted onto nitrocellulose membranes and air-dried. The membranes were blocked with 5% skim milk, then incubated with a primary antibody against 8-Hydroxyguanosine overnight at 4°C. After washing, the membranes were incubated with a horseradish peroxidase-conjugated secondary antibody. The signal was visualized using a chemiluminescence reagent, and the intensity was quantified by densitometry. The concentration of 8-Hydroxyguanosine was determined by comparing with a standard curve [2] |
| Cell Assay |
Murine splenic B lymphocyte culture and immunostimulation assay: Spleens were removed from BALB/c mice under sterile conditions and minced into small pieces. The tissue was digested with collagenase for 15 minutes, and a single-cell suspension was prepared by passing through a 200 μm mesh. B lymphocytes were isolated using nylon wool columns (retaining T lymphocytes, allowing B lymphocytes to pass through). Isolated B cells were adjusted to a concentration of 2×10^6 cells/mL and seeded in 96-well plates (100 μL per well). 8-Hydroxyguanosine was added at final concentrations of 10 μg/mL, 20 μg/mL, 40 μg/mL, and 80 μg/mL. For proliferation detection, 0.5 μCi of [3H]-thymidine was added to each well 18 hours before the end of the 72-hour incubation, and radioactivity was measured using a liquid scintillation counter. For antibody detection, culture supernatant was collected after 72 hours, and IgG/IgM levels were measured by ELISA [3]
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| Animal Protocol |
Rat liver oxidative damage model (2-nitropropane-induced): Male Sprague-Dawley rats (180-220 g) were housed under controlled conditions (12-hour light/dark cycle, 22±2°C) with free access to food and water. Rats were randomly divided into 2 groups (n=6/group): Control group (administered with physiological saline) and 2-nitropropane-treated group (administered with 2-nitropropane). 2-Nitropropane was dissolved in physiological saline to a concentration of 50 mg/mL, and administered via oral gavage at a dose of 100 mg/kg body weight, twice daily (8:00 and 18:00) for 14 consecutive days. On day 15, rats were sacrificed by cervical dislocation under anesthesia. Livers were quickly excised, rinsed with cold physiological saline, and stored at -80°C until DNA/RNA extraction and 8-Hydroxyguanosine detection [1]
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| References |
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| Additional Infomation |
8-Hydroxyguanosine is a purine nucleoside.
1. 8-Hydroxyguanosine is a major oxidative stress biomarker formed by the oxidation of guanosine residues in DNA and RNA, and is widely used to assess oxidative damage to nucleic acids under various pathological conditions (e.g., carcinogenicity, neurodegenerative diseases) [1][2] 2. In the literature [1], 8-Hydroxyguanosine was used as an indicator of liver DNA/RNA oxidative damage induced by the hepatocarcinogen 2-nitropropane; its elevated level directly reflects the degree of oxidative stress and nucleic acid damage in liver tissue [1] 3. The literature [2] reported that the concentration of 8-Hydroxyguanosine in the cerebrospinal fluid of AD patients was elevated, suggesting that oxidative stress and nucleic acid damage are involved in the pathogenesis of AD, supporting 8-Hydroxyguanosine as a potential diagnostic biomarker for AD [2] 4. The literature [3] is the only study to report the biological activity of 8-Hydroxyguanosine: it exerts an immunostimulatory effect on B lymphocytes by promoting cell proliferation and enhancing antibody secretion, indicating that it may play a role in regulating humoral immunity [3] |
| Molecular Formula |
C10H13N5O6
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|---|---|
| Molecular Weight |
299.24012
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| Exact Mass |
299.086
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| CAS # |
3868-31-3
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| PubChem CID |
135407175
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| Appearance |
Typically exists as solid at room temperature
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| Density |
2.4±0.1 g/cm3
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| Melting Point |
232-235ºC
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| Index of Refraction |
2.007
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| LogP |
-0.97
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| Hydrogen Bond Donor Count |
6
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
21
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| Complexity |
574
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| Defined Atom Stereocenter Count |
4
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| SMILES |
OC[C@@H]1[C@H]([C@H]([C@H](N2C(NC3=C2N=C(N)NC3=O)=O)O1)O)O
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| InChi Key |
FPGSEBKFEJEOSA-UMMCILCDSA-N
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| InChi Code |
InChI=1S/C10H13N5O6/c11-9-13-6-3(7(19)14-9)12-10(20)15(6)8-5(18)4(17)2(1-16)21-8/h2,4-5,8,16-18H,1H2,(H,12,20)(H3,11,13,14,19)/t2-,4-,5-,8-/m1/s1
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| Chemical Name |
2-amino-9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-1,7-dihydropurine-6,8-dione
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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) |
DMSO : ~125 mg/mL (~417.72 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.95 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (6.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (6.95 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.3418 mL | 16.7090 mL | 33.4180 mL | |
| 5 mM | 0.6684 mL | 3.3418 mL | 6.6836 mL | |
| 10 mM | 0.3342 mL | 1.6709 mL | 3.3418 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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