- Cell Adhesion Molecules (CAMs), including Intercellular Adhesion Molecule-1 (ICAM-1) and Vascular Cell Adhesion Molecule-1 (VCAM-1) [1]
- The protective effect was associated with modulation of oxidative stress markers (e.g., Superoxide Dismutase (SOD), Malondialdehyde (MDA)), inflammatory cytokines (e.g., Tumor Necrosis Factor-α (TNF-α), Interleukin-6 (IL-6)), and apoptotic proteins (e.g., Bax, Bcl-2, Caspase-3) [2]
- Toll-like Receptor 2 (TLR2), p38 Mitogen-Activated Protein Kinase (p38 MAPK), and Nuclear Factor-kappa B (NF-κB) [3]

- In brain microvascular endothelial cells (BMECs) subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) injury: Pretreatment with Polydatin (10, 30, 100 μM) dose-dependently reduced the expression of ICAM-1 and VCAM-1 at both mRNA (detected by RT-PCR) and protein (detected by western blot) levels. Additionally, Polydatin (30, 100 μM) significantly inhibited the adhesion of polymorphonuclear leukocytes (PMNs) to OGD/R-injured BMECs (the adhesion rate was reduced by approximately 35% and 50% at 30 μM and 100 μM, respectively, compared to the OGD/R group) [1]
- In H9c2 cardiomyocytes or primary rat cardiomyocytes injured by doxorubicin (DOX, 1 μM): Co-treatment with Polydatin (5, 10, 20 μM) dose-dependently increased cell viability (detected by MTT assay; cell viability was increased by ~20%, ~35%, and ~50% at 5 μM, 10 μM, and 20 μM, respectively, compared to the DOX group), elevated SOD activity and glutathione peroxidase (GSH-Px) activity, and decreased MDA content and intracellular reactive oxygen species (ROS) levels (detected by DCFH-DA staining). Furthermore, Polydatin (10, 20 μM) downregulated the mRNA and protein expression of TNF-α and IL-6 (detected by RT-PCR and ELISA), reduced Bax protein expression, increased Bcl-2 protein expression, and inhibited Caspase-3 activation (detected by western blot). When combined with Vitamin C (VC, 50 μM), the protective effects of Polydatin (10 μM) were significantly enhanced (e.g., cell viability was further increased by ~15% compared to Polydatin alone) [2]
- In MAC-T bovine mammary epithelial cells infected with Staphylococcus aureus (S. aureus, MOI = 10:1): Treatment with Polydatin (25, 50, 100 μM) dose-dependently improved cell viability (detected by CCK-8 assay; cell viability was increased by ~18%, ~32%, and ~45% at 25 μM, 50 μM, and 100 μM, respectively, compared to the S. aureus-infected group), decreased the secretion of IL-1β, IL-6, and TNF-α (detected by ELISA; IL-6 levels were reduced by ~30%, ~48%, and ~65% at the three concentrations, respectively), and downregulated the mRNA expression of these cytokines (detected by RT-PCR). Additionally, Polydatin (50, 100 μM) inhibited S. aureus-induced phosphorylation of p38 MAPK and NF-κB p65, and reduced TLR2 protein expression (detected by western blot). Immunofluorescence analysis showed that Polydatin (100 μM) suppressed the nuclear translocation of NF-κB p65 in infected MAC-T cells [3]

- In Sprague-Dawley (SD) rats with middle cerebral artery occlusion/reperfusion (MCAO/R) injury: Intraperitoneal (i.p.) administration of Polydatin (20, 40 mg/kg) 30 minutes before MCAO significantly reduced cerebral infarct volume (detected by TTC staining; infarct volume was reduced by ~28% and ~42% at 20 mg/kg and 40 mg/kg, respectively, compared to the MCAO/R group) and improved neurological function (evaluated by Longa scoring; neurological scores were decreased by ~1.5 and ~2.2 points at the two doses, respectively). Histopathological examination (HE staining) showed that Polydatin (40 mg/kg) reduced leukocyte infiltration in the ischemic brain tissue. Western blot and immunohistochemical analysis revealed that Polydatin (20, 40 mg/kg) dose-dependently downregulated the protein expression of ICAM-1 and VCAM-1 in the cerebral cortex and hippocampus [1]
- In Wistar rats with DOX-induced cardiotoxicity (DOX: i.p. 2.5 mg/kg, twice a week for 4 weeks, total dose 20 mg/kg): Oral gavage (i.g.) administration of Polydatin (10, 20 mg/kg) once daily for 5 weeks (starting 1 week before DOX administration) dose-dependently reduced serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) (CK-MB levels were reduced by ~25% and ~40% at 10 mg/kg and 20 mg/kg, respectively, compared to the DOX group), increased myocardial SOD and GSH-Px activities, and decreased myocardial MDA content. Histopathological analysis (HE staining) showed that Polydatin (20 mg/kg) alleviated DOX-induced myocardial necrosis and inflammatory cell infiltration. TUNEL assay and western blot demonstrated that Polydatin (20 mg/kg) reduced myocardial cell apoptosis (apoptosis rate was decreased by ~30%), downregulated Bax and cleaved-Caspase-3 protein expression, and upregulated Bcl-2 protein expression. Co-administration of Polydatin (20 mg/kg, i.g.) and VC (100 mg/kg, i.g.) showed a synergistic effect, as evidenced by further reductions in serum CK-MB (additional ~12% reduction compared to Polydatin alone) and myocardial apoptosis rate (additional ~10% reduction) [2]
- In lactating BALB/c mice with S. aureus-induced mastitis (S. aureus: 1×10^6 CFU per mouse, injected into the mammary duct): Intraperitoneal (i.p.) administration of Polydatin (10, 20, 40 mg/kg) at 1 h and 12 h post-infection dose-dependently decreased the bacterial load in mammary tissue (detected by CFU counting; CFU was reduced by ~35%, ~55%, and ~70% at 10 mg/kg, 20 mg/kg, and 40 mg/kg, respectively, compared to the infected group), alleviated mammary tissue damage (HE staining showed reduced alveolar destruction and inflammatory cell infiltration), and decreased the levels of IL-1β, IL-6, and TNF-α in mammary tissue (detected by ELISA and RT-PCR; IL-1β levels were reduced by ~32%, ~50%, and ~68% at the three doses, respectively). Western blot and immunohistochemical analysis indicated that Polydatin (20, 40 mg/kg) dose-dependently inhibited S. aureus-induced upregulation of TLR2 protein expression, and phosphorylation of p38 MAPK and NF-κB p65 in mammary tissue [3]

- Assay for SOD, GSH-Px, and MDA in myocardial tissue/cells: Tissue or cell homogenates were prepared and centrifuged to obtain supernatants. For SOD activity assay: Supernatant was mixed with reaction buffer containing xanthine and xanthine oxidase, and the absorbance was measured at 550 nm after incubation; SOD activity was calculated based on the inhibition rate of superoxide anion-induced nitrite formation. For GSH-Px activity assay: Supernatant was incubated with GSH and hydrogen peroxide, and the remaining GSH was measured by reaction with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) at 412 nm; GSH-Px activity was calculated based on the consumption rate of GSH. For MDA assay: Supernatant was mixed with thiobarbituric acid (TBA) and heated in a boiling water bath; the absorbance of the resulting MDA-TBA adduct was measured at 532 nm, and MDA content was calculated using a standard curve [2]
- Assay for NF-κB activity in MAC-T cells (luciferase reporter gene assay): MAC-T cells were transfected with NF-κB luciferase reporter plasmid and Renilla luciferase plasmid (internal control) using transfection reagent. After transfection, cells were infected with S. aureus and treated with Polydatin (100 μM) for 6 h. Cells were lysed, and luciferase activity was measured using a dual-luciferase reporter assay system; NF-κB activity was expressed as the ratio of firefly luciferase activity to Renilla luciferase activity [3]

- OGD/R model and cell adhesion assay in BMECs: BMECs were cultured in high-glucose DMEM medium. For OGD treatment: Cells were washed with glucose-free DMEM and incubated in glucose-free DMEM in a hypoxic chamber (95% N2, 5% CO2) at 37°C for 4 h, followed by reoxygenation in high-glucose DMEM under normoxic conditions (95% air, 5% CO2) for 24 h. For Polydatin treatment: Cells were pretreated with Polydatin (10, 30, 100 μM) for 2 h before OGD. For cell adhesion assay: PMNs were labeled with fluorescein isothiocyanate (FITC), then co-incubated with treated BMECs for 1 h at 37°C. Non-adherent PMNs were washed away, and the fluorescence intensity of adherent PMNs was measured using a microplate reader to calculate the adhesion rate [1]
- DOX-induced injury assay in H9c2 cells: H9c2 cells were cultured in DMEM medium containing 10% fetal bovine serum. Cells were treated with DOX (1 μM) alone or in combination with Polydatin (5, 10, 20 μM) and/or VC (50 μM) for 24 h. Cell viability was detected by MTT assay: MTT solution (5 mg/mL) was added to each well and incubated for 4 h, then the supernatant was removed, and dimethyl sulfoxide (DMSO) was added to dissolve the formazan crystals; the absorbance was measured at 490 nm. Intracellular ROS levels were detected by DCFH-DA staining: Cells were incubated with DCFH-DA (10 μM) for 30 min, washed, and the fluorescence intensity was measured by flow cytometry or fluorescence microscopy [2]
- S. aureus infection assay in MAC-T cells: MAC-T cells were cultured in RPMI-1640 medium. Cells were infected with S. aureus (MOI = 10:1) for 1 h, then washed to remove unbound bacteria, and treated with Polydatin (25, 50, 100 μM) for 23 h. Cell viability was detected by CCK-8 assay: CCK-8 solution was added to each well and incubated for 2 h, then the absorbance was measured at 450 nm. For NF-κB nuclear translocation assay: Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with bovine serum albumin, incubated with anti-NF-κB p65 primary antibody and FITC-conjugated secondary antibody, and stained with DAPI for nuclei; the localization of NF-κB p65 was observed under a confocal laser scanning microscope [3]

- MCAO/R model and Polydatin administration in SD rats: Male SD rats (250-300 g) were anesthetized with chloral hydrate (350 mg/kg, i.p.). The right middle cerebral artery (MCA) was occluded using a nylon suture (0.26 mm diameter) inserted through the external carotid artery into the internal carotid artery until resistance was felt (approximately 18-20 mm from the carotid bifurcation). After 2 h of occlusion, the suture was withdrawn to allow reperfusion. Rats were randomly divided into 4 groups (n = 6-8 per group): Sham operation group (no MCA occlusion, i.p. injection of normal saline), MCAO/R group (MCAO/R + i.p. normal saline), Polydatin 20 mg/kg group (MCAO/R + i.p. Polydatin 20 mg/kg), Polydatin 40 mg/kg group (MCAO/R + i.p. Polydatin 40 mg/kg). Polydatin was dissolved in normal saline containing 0.5% DMSO. The neurological function of rats was evaluated using the Longa scoring system (0-4 points) at 24 h after reperfusion. Rats were then euthanized, and the brains were removed. One part of the brain was cut into 2 mm coronal slices, stained with 2% TTC at 37°C for 30 min, and photographed; the infarct volume was analyzed using ImageJ software. Another part of the brain tissue (cerebral cortex and hippocampus) was homogenized for western blot analysis, and the remaining part was fixed with 4% paraformaldehyde for immunohistochemical staining [1]
- DOX-induced cardiotoxicity model and Polydatin/VC administration in Wistar rats: Male Wistar rats (200-250 g) were randomly divided into 6 groups (n = 6 per group): Normal control group (i.g. normal saline + i.p. normal saline), DOX group (i.g. normal saline + i.p. DOX), Polydatin 10 mg/kg group (i.g. Polydatin 10 mg/kg + i.p. DOX), Polydatin 20 mg/kg group (i.g. Polydatin 20 mg/kg + i.p. DOX), VC 100 mg/kg group (i.g. VC 100 mg/kg + i.p. DOX), Polydatin 20 mg/kg + VC 100 mg/kg group (i.g. Polydatin 20 mg/kg + i.g. VC 100 mg/kg + i.p. DOX). Polydatin was dissolved in normal saline containing 0.5% Tween 80, and VC was dissolved in normal saline. DOX was administered i.p. at 2.5 mg/kg twice a week for 4 weeks (total dose 20 mg/kg). Polydatin and VC were administered i.g. once daily for 5 weeks, starting 1 week before the first DOX injection. At the end of the experiment, rats were anesthetized with ether, and blood was collected from the abdominal aorta to separate serum for measuring CK-MB and LDH levels using commercial biochemical kits. Rats were then euthanized, and the heart was removed, weighed, and the heart weight index (heart weight/body weight × 1000) was calculated.

[1]. Involvement of cell adhesion molecules in polydatin protection of brain tissues from ischemia-reperfusion injury. Brain Res. 2006 Sep 19;1110(1):193-200.

[2]. Synergistic effects of Polydatin and Vitamin C in Inhibiting Cardiotoxicity induced by Doxorubicin in rats. Fundam Clin Pharmacol. 2016 Nov 28. [Epub ahead of print].

[3]. Polydatin ameliorates Staphylococcus aureus-induced mastitis in mice via inhibiting TLR2-mediated activation of the p38 MAPK/NF-κB pathway. Acta Pharmacol Sin. 2016 Nov 28. [Epub ahead of print].

[4]. A New Inhibitor of glucose-6-phosphate Dehydrogenase Blocks Pentose Phosphate Pathway and Suppresses Malignant Proliferation and Metastasis in Vivo. Cell Death Dis. 2018 May 1;9(5):572.

Trans-piceid is a stilbene compound, a product of trans-resveratrol with a β-D-glucose residue at the 3-position. It has multiple functions, including as a metabolite, potassium channel regulator, antiarrhythmic drug, hepatoprotective agent, antioxidant, nephroprotective agent, and anti-aging agent. It is a stilbene compound, polyphenol, β-D-glucoside, and monosaccharide derivative. Its function is related to trans-resveratrol. Polydatin, also known as Piceid, is the natural precursor and glycosidic form of resveratrol, possessing a single-crystal structure. Although it is isolated from the bark of Picea sitchensis or Polygonum cuspidatum, resveratrol glycosides may also be found in grapes, peanuts, hop cones, red wine, hop kernels, cocoa products, chocolate products, and many everyday foods. Resveratrol glycosides possess anti-inflammatory, immunomodulatory, antioxidant, and antitumor activities. Studies have shown that they can exert cytotoxic effects on colorectal cancer cells by inducing cell cycle arrest and apoptosis. Resveratrol glycosides have been reported to be found in hops, resveratrol, and other organisms with relevant data.

C20H22O8

390.3839

390.131

27208-80-6

Polydatin;27208-80-6

5281718

White to off-white solid powder

1.5±0.1 g/cm3

707.7±60.0 °C at 760 mmHg

130 - 140 °C

381.8±32.9 °C

0.0±2.4 mmHg at 25°C

1.737

1

6

8

5

28

506

5

C1=CC(=CC=C1/C=C/C2=CC(=CC(=C2)O[C@H]3[C@@H]([C@H]([C@@H]([C@H](O3)CO)O)O)O)O)O

HSTZMXCBWJGKHG-CUYWLFDKSA-N

InChI=1S/C20H22O8/c21-10-16-17(24)18(25)19(26)20(28-16)27-15-8-12(7-14(23)9-15)2-1-11-3-5-13(22)6-4-11/h1-9,16-26H,10H2/b2-1+/t16-,17-,18+,19-,20-/m1/s1

(2S,3R,4S,5S,6R)-2-[3-hydroxy-5-[(E)-2-(4-hydroxyphenyl)ethenyl]phenoxy]-6-(hydroxymethyl)oxane-3,4,5-triol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 29 mg/mL (~74.29 mM)
H2O : ~0.1 mg/mL (~0.26 mM)

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)