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| ln Vitro |
Puerarin 6″-O-Xyloside (PXY) promoted the proliferation of osteoblasts isolated from neonatal rat calvarias in a dose- and time-dependent manner. Treatment with 5, 10, and 20 µM PXY for 24, 48, and 72 hours significantly increased the optical density (OD) at 550 nm in the MTT assay compared to the control group. [1]
Western blot analysis demonstrated that Puerarin 6″-O-Xyloside (PXY) regulated the expression of key proteins involved in bone metabolism in cultured osteoblasts. Treatment with 10 and 20 µM PXY for 48 hours significantly up-regulated the expression of osteoprotegerin (OPG) protein. Conversely, treatment with 5, 10, and 20 µM PXY significantly down-regulated the expression of receptor activator of NF-κB ligand (RANKL) protein. Consequently, PXY significantly increased the OPG/RANKL ratio. [1] |
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| ln Vivo |
In ovariectomized (OVX) mice, intraperitoneal administration of Puerarin 6″-O-Xyloside (PXY) at doses of 40 and 60 mg/kg/day for 12 weeks significantly reduced the increase in body weight induced by OVX. [1]
PXY treatment (20, 40, and 60 mg/kg/day, i.p., 12 weeks) significantly increased the uterine index (uterus weight/body weight) in OVX mice in a dose-dependent manner, ameliorating the uterine atrophy caused by estrogen deficiency. [1] PXY treatment (20, 40, and 60 mg/kg/day, i.p., 12 weeks) significantly increased the lowered serum levels of blood calcium and blood phosphorus in OVX mice in a dose-dependent manner. [1] PXY treatment at doses of 40 and 60 mg/kg/day (i.p., 12 weeks) significantly increased the serum alkaline phosphatase (ALP) activity in OVX mice. [1] PXY treatment (20, 40, and 60 mg/kg/day, i.p., 12 weeks) significantly increased the reduced serum osteoprotegerin (OPG) levels in OVX mice in a dose-dependent manner. [1] Histopathological examination of femoral bone tissue showed that OVX caused bone loss, characterized by enlarged bone marrow cavity, sparse and fractured trabecular bone. Treatment with PXY (20, 40, and 60 mg/kg/day, i.p., 12 weeks) improved these osteoporotic changes, resulting in thickened compact bone and denser trabecular bone. The improvement was most pronounced at the 60 mg/kg dose. [1] |
| Cell Assay |
Osteoblasts were isolated from calvarias of 3-day-old neonatal rats. Briefly, calvarial bones were minced and sequentially digested with trypsin and then with a solution containing trypsin and collagenase type II. The released cells were collected by centrifugation and cultured in α-MEM supplemented with fetal bovine serum. [1]
For the proliferation assay, osteoblasts were seeded in 96-well plates and treated with various concentrations of PXY (5, 10, 20 µM) for 24, 48, or 72 hours. Cell viability/proliferation was assessed using the MTT assay. After the incubation period, MTT reagent was added to each well. Following a further incubation, the formazan crystals were dissolved, and the optical density was measured at 550 nm using a microplate reader. [1] For western blot analysis, osteoblasts were treated with or without PXY (5, 10, 20 µM) for 48 hours. Cells were then harvested, and total proteins were extracted. Equal amounts of protein were separated by SDS-PAGE, transferred to a PVDF membrane, and probed with primary antibodies against RANKL and OPG. After incubation with an appropriate horseradish peroxidase-conjugated secondary antibody, protein bands were visualized using a chemiluminescence detection system. β-actin was used as a loading control. [1] |
| Animal Protocol |
An osteoporotic mouse model was established by bilateral ovariectomy (OVX) in 3-month-old female ICR mice. Sham-operated mice underwent the same surgical procedure without ovary removal. [1]
One week after surgery, OVX mice were randomly divided into groups (n=10): OVX model group, and PXY treatment groups (20, 40, 60 mg/kg/day). A sham-operated group was also included. [1] Puerarin 6″-O-Xyloside (PXY) was administered via intraperitoneal injection (i.p.) daily for 12 consecutive weeks. The vehicle control group received an equivalent volume of normal saline (10 ml/kg, i.p.). [1] At the end of the 12-week treatment period, body weights were recorded. Mice were then euthanized. Blood was collected to obtain serum for biochemical analysis. Uteri were removed and weighed to calculate the uterine index. Femurs were collected for histopathological examination after fixation, decalcification, paraffin embedding, sectioning, and H&E staining. [1] |
| Toxicity/Toxicokinetics |
Acute toxicity tests were conducted in ICR mice. Mice in each group received a single intraperitoneal injection of 10, 20, 40, 60, or 80 mg/kg of puerarin 6″-O-xyloside (PXY). Mice were observed for mortality and any abnormal neurological behavior within 24 hours. No death or abnormal behavior was observed at any of the tested doses. Since no significant toxicity was observed within the tested dose range, the median lethal dose (LD50) could not be determined. [1]
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| References |
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| Additional Infomation |
Puerarin 6″-O-xyloside (PXY) is the main isoflavone component in kudzu root.[1] Studies have shown that the anti-osteoporosis activity of PXY may exert its effects through multiple mechanisms, including increasing serum calcium, phosphorus and alkaline phosphatase (ALP) levels, and upregulating the osteoprotegerin/nuclear factor activator receptor L (OPG/RANKL) ratio in osteoblasts, which is a key pathway regulating osteoclast formation and bone resorption.[1]
|
| Molecular Formula |
C26H28O13
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|---|---|
| Molecular Weight |
548.4927
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| Exact Mass |
548.153
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| CAS # |
114240-18-5
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| PubChem CID |
100990912
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| Appearance |
White to off-white solid powder
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| Density |
1.7±0.1 g/cm3
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| Boiling Point |
833.0±65.0 °C at 760 mmHg
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| Flash Point |
280.6±27.8 °C
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| Vapour Pressure |
0.0±3.2 mmHg at 25°C
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| Index of Refraction |
1.746
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| LogP |
1.48
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| Hydrogen Bond Donor Count |
8
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| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
39
|
| Complexity |
891
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| Defined Atom Stereocenter Count |
9
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| SMILES |
C1[C@H]([C@@H]([C@H]([C@@H](O1)OC[C@@H]2[C@H]([C@@H]([C@H]([C@@H](O2)C3=C(C=CC4=C3OC=C(C4=O)C5=CC=C(C=C5)O)O)O)O)O)O)O)O
|
| InChi Key |
YCFQXQCEEPCZMO-KATYHMCLSA-N
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| InChi Code |
InChI=1S/C26H28O13/c27-11-3-1-10(2-4-11)13-7-36-24-12(18(13)30)5-6-14(28)17(24)25-22(34)21(33)20(32)16(39-25)9-38-26-23(35)19(31)15(29)8-37-26/h1-7,15-16,19-23,25-29,31-35H,8-9H2/t15-,16-,19+,20-,21+,22-,23-,25+,26+/m1/s1
|
| Chemical Name |
7-hydroxy-3-(4-hydroxyphenyl)-8-[(2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-[[(2S,3R,4S,5R)-3,4,5-trihydroxyoxan-2-yl]oxymethyl]oxan-2-yl]chromen-4-one
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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) |
DMSO : ~100 mg/mL (~182.32 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.56 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 25.0 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: ≥ 2.5 mg/mL (4.56 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 25.0 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: ≥ 2.5 mg/mL (4.56 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 | 1.8232 mL | 9.1159 mL | 18.2319 mL | |
| 5 mM | 0.3646 mL | 1.8232 mL | 3.6464 mL | |
| 10 mM | 0.1823 mL | 0.9116 mL | 1.8232 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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