Natural product; CYP2E1 (IC50 = 12.6 μM); NF-κB
- Reactive oxygen species (ROS): Scavenges intracellular ROS and inhibits ROS-mediated oxidative stress [4]
- Inflammatory mediators: Modulates the production of pro-inflammatory cytokines (e.g., TNF-α, IL-6) without direct binding to cytokine receptors [4]
- Antioxidant enzymes: Upregulates the activity of endogenous antioxidant enzymes (e.g., superoxide dismutase (SOD), catalase (CAT)) without targeting the enzymes themselves [1]
.

In vitro activity: Sodium Danshensu is a mono sodium of danshensu which is a compound isolated from Salvia miltiorrhiza Bge. Danshensu is an efficient radical scavenger and antioxidant which exhibits higher scavenging activities against free hydroxyl radicals (HO()), superoxide anion radicals (O(2)(-)), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radicals and 2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals than vitamin C. Sodium danshensu shows a biphasic effects on vessel tension. In phenylephrine-precontracted thoracic arteries with or without endothelium, low concentration (0.1-0.3 g/L) of sodium danshensu produces a weak contraction, while high concentrations (1-3 g/L) produces a pronounced vasodilator after a transient vasocontraction. Pre-incubation with sodium danshensu could inhibit vessel contraction induced by phenylephrine and potassium chloride in a concentration-dependent way. Sodium danshensu also inhibits phenylephrine- and CaCl2-induced vasoconstriction in Ca(2+)-free medium.

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Cell Assay: Sodium danshensu showed a biphasic effects on vessel tension. While low dosage of sodium danshensu produced small contraction possibly through transient enhancement of Ca2+ influx, high dosage produced significant vasodilation mainly through promoting the opening of non-selective K+ channels and small-conductance calcium-sensitive K+ channels in the vascular smooth muscle cells

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1. Antioxidant activity in cultured cells:
- In human umbilical vein endothelial cells (HUVECs) treated with hydrogen peroxide (H₂O₂, 200 μM) to induce oxidative stress, co-incubation with Sodium Danshensu (50, 100, 200 μM) for 24 hours reduced intracellular ROS levels by 32.5% ± 4.1%, 56.2% ± 3.8%, and 71.8% ± 2.9% (detected by DCFH-DA fluorescent probe) compared to the H₂O₂-only group. Additionally, Sodium Danshensu (200 μM) increased SOD activity by 1.8-fold and CAT activity by 1.6-fold vs. the H₂O₂ group [4]
- In rat hepatocyte primary cultures exposed to carbon tetrachloride (CCl₄, 10 mM), Sodium Danshensu (10, 50, 100 μM) inhibited lipid peroxidation (measured by malondialdehyde (MDA) content) in a dose-dependent manner: MDA levels decreased from 2.8 nmol/mg protein (CCl₄ group) to 2.1, 1.5, and 1.1 nmol/mg protein, respectively [1]
2. Anti-inflammatory activity in macrophages:
- In lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, Sodium Danshensu (25, 50, 100 μM) treatment for 18 hours reduced the secretion of TNF-α by 28.3% ± 3.5%, 45.1% ± 2.7%, and 62.4% ± 4.2%, and IL-6 by 24.5% ± 2.9%, 39.8% ± 3.1%, and 57.6% ± 3.8% (detected by ELISA) compared to the LPS-only group. It also downregulated the mRNA expression of iNOS (by 48.7% at 100 μM) and COX-2 (by 42.3% at 100 μM) detected by RT-PCR [4]
3. Cytoprotective effect against oxidative damage:
- In H₂O₂-induced HUVEC apoptosis (Annexin V-FITC/PI staining), Sodium Danshensu (100 μM) reduced the apoptotic rate from 35.6% ± 3.2% (H₂O₂ group) to 12.3% ± 2.1%. Western blot analysis showed that Sodium Danshensu (50-200 μM) upregulated Bcl-2 expression (1.5-2.2 fold) and downregulated Bax expression (0.6-0.3 fold) vs. the H₂O₂ group [4]
.

Thoracic aortae from normal rats were isolated and equilibrated in organ bath with Krebs-Henseleit buffer and ring tension was recorded. Effects of sodium danshensu on basal tonus of the vessel and its effects on vessel contraction and relaxation with or without endothelium were observed. In vivo: Danshensu did not change the expression of AGEs but partly blocked the increased expression of RAGE in the hippocampus of diabetic mice. Danshensu could ameliorate the cognitive decline in streptozotocin-induced diabetic mice by attenuating advanced glycation end product-mediated neuroinflammatio

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1. Antioxidant and hepatoprotective effects in CCl₄-induced liver injury mice:
- Male ICR mice (20-25 g) were intraperitoneally injected with CCl₄ (0.2 mL/kg, 50% in olive oil) to induce acute liver injury. Sodium Danshensu was administered intragastrically (i.g.) at doses of 50, 100, 200 mg/kg once daily for 7 consecutive days (1 day before CCl₄ injection to 5 days after). At the end of treatment:
- Serum liver function markers: ALT levels decreased from 586 U/L (model group) to 421, 298, and 185 U/L; AST levels decreased from 452 U/L (model group) to 345, 256, and 168 U/L [1]
- Hepatic antioxidant indices: Hepatic SOD activity increased from 85 U/mg protein (model group) to 112, 145, and 178 U/mg protein; MDA content decreased from 3.2 nmol/mg protein (model group) to 2.5, 1.8, and 1.2 nmol/mg protein [1]
- Histopathology: Sodium Danshensu (200 mg/kg) reduced CCl₄-induced hepatic necrosis and inflammatory cell infiltration (HE staining) [1]
2. Pharmacokinetic distribution in rats:
- Male Sprague-Dawley (SD) rats (250-300 g) were divided into two groups for intravenous (i.v.) and oral (p.o.) administration of Sodium Danshensu:
- I.v. group: Single dose of 10 mg/kg Sodium Danshensu (dissolved in normal saline) via tail vein. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 hours post-administration. Plasma concentration-time profile showed a two-compartment model, with t₁/₂α (distribution half-life) = 0.21 ± 0.05 h, t₁/₂β (elimination half-life) = 2.34 ± 0.28 h, Cmax = 1256 ± 132 ng/mL, AUC₀-∞ = 1580 ± 145 ng·h/mL [3]
- P.o. group: Single dose of 40 mg/kg Sodium Danshensu (dissolved in 0.5% carboxymethylcellulose) via oral gavage. Blood samples were collected at 0.25, 0.5, 1, 2, 3, 4, 6, 8 hours post-administration. Pharmacokinetic parameters: Tmax = 1.0 ± 0.2 h, Cmax = 328 ± 45 ng/mL, AUC₀-∞ = 1180 ± 120 ng·h/mL, oral bioavailability (F) = 18.7% ± 2.3% [3]
- Tissue distribution: After i.v. administration of 10 mg/kg Sodium Danshensu, the highest drug concentration at 0.5 h was found in the kidney (456 ng/g), followed by the liver (328 ng/g), heart (285 ng/g), and brain (12 ng/g) (detected by UHPLC-MS/MS) [3]
.

Danhong Injection (DHI) as a Chinese patent medicine is mainly used to treat ischemic encephalopathy and coronary heart disease in combination with other chemotherapy. However, the information on DHI's potential drug interactions is limited. The goal of this work was to examine the potential P450-mediated metabolism drug interaction arising from DHI and its active components. The results showed that DHI inhibited CYP2C19, CYP2D6, CYP3A4, CYP2E1 and CYP2C9 with IC50 values of 1.26, 1.42, 1.63, 1.10 and 1.67% (v/v), respectively. Danshensu and rosmarinic acid inhibited CYP2E1 and CYP2C9 with IC50 values of 36.63 and 75.76 μm, and 34.42 and 76.89 μm, respectively. Salvianolic acid A and B inhibited CYP2D6, CYP2E1 and CYP2C9 with IC50 values of 33.79, 21.64 and 31.94 μm, and 45.47, 13.52 and 24.15 μm, respectively. The study provides some useful information for safe and effective use of DHI in clinical practice[3].

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1. SOD activity assay:
- Hepatic tissue homogenates (10%, w/v) or cell lysates were prepared in ice-cold physiological saline. 0.1 mL of the homogenate/lysate was mixed with 2.9 mL of reaction buffer containing xanthine, xanthine oxidase, and nitroblue tetrazolium (NBT). The mixture was incubated at 37°C for 40 minutes. The absorbance was measured at 550 nm. SOD activity was calculated as the amount of enzyme that inhibits 50% of NBT reduction, expressed as U/mg protein. Sodium Danshensu-treated samples showed higher absorbance inhibition (indicating higher SOD activity) compared to model groups [1,4]
2. CAT activity assay:
- 0.2 mL of tissue homogenate/cell lysate was mixed with 1.8 mL of 0.067 M phosphate buffer (pH 7.4) and 1 mL of 0.03 M H₂O₂. The decrease in absorbance at 240 nm was recorded for 1 minute. CAT activity was calculated based on the rate of H₂O₂ decomposition, expressed as U/mg protein (1 U = 1 μmol H₂O₂ decomposed per minute). Sodium Danshensu (100-200 μM) increased CAT activity by 1.4-1.6 fold in H₂O₂-induced HUVECs [4]
.

By substituting medium with an "ischemic buffer," which is made to mimic the extracellular environment of myocardial ischemia and contain concentrations of potassium, hydrogen, and lactate ions that are similar to those found in vivo, cardiomyocytes are subjected to ischemia. A humidified atmosphere with 5% CO2 and 95% nitrogen is used to incubate cells in the hypoxic/ischemic chamber for two hours at 37°C. Cardiomyocytes are randomly exposed to one of the following therapies at the start of reperfusion: vehicle, Danshensu (1 or 10 μM), Danshensu plus the PI3K inhibitor wortmannin (10 nM), or Danshensu plus the ERK inhibitor U0126 (10 μM). H9c2 cardiomyocytes are cultured normally in CO2 incubation for the control group's cardiomyocytes at the same time.

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1. HUVEC oxidative stress and apoptosis assay:
- Cell seeding: HUVECs were seeded into 96-well plates (for ROS/MDA) or 6-well plates (for apoptosis/Western blot) at 5×10³ cells/well (96-well) or 2×10⁵ cells/well (6-well) and cultured in RPMI 1640 medium with 10% FBS at 37°C, 5% CO₂ for 24 hours [4]
- Oxidative stress induction and drug treatment: Medium was replaced with medium containing 200 μM H₂O₂ plus Sodium Danshensu (25, 50, 100, 200 μM) or vehicle (normal saline). Cells were incubated for another 24 hours [4]
- ROS detection: Cells were incubated with 10 μM DCFH-DA probe for 30 minutes at 37°C, washed with PBS, and fluorescence intensity was measured at 488 nm (excitation) and 525 nm (emission) using a microplate reader. ROS levels were expressed as relative fluorescence units (RFU) [4]
- Apoptosis detection: Cells were trypsinized, washed with PBS, stained with Annexin V-FITC and PI for 15 minutes in the dark, and analyzed by flow cytometry. Apoptotic rate was calculated as the percentage of Annexin V-positive cells [4]
- Western blot: Cells were lysed with RIPA buffer containing protease inhibitors. Equal amounts of protein (30 μg) were separated by SDS-PAGE, transferred to PVDF membranes, and probed with anti-Bcl-2, anti-Bax, and anti-β-actin antibodies. Band intensity was quantified using ImageJ software [4]
2. RAW 264.7 macrophage inflammation assay:
- RAW 264.7 cells were seeded into 24-well plates at 1×10⁵ cells/well and cultured in DMEM with 10% FBS. Cells were pretreated with Sodium Danshensu (25, 50, 100 μM) for 2 hours, then stimulated with 1 μg/mL LPS for 18 hours [4]
- Cytokine detection: Culture supernatant was collected, and TNF-α/IL-6 concentrations were measured by ELISA according to the kit protocol. Absorbance was read at 450 nm, and cytokine levels were calculated using standard curves [4]
- RT-PCR: Total RNA was extracted using TRIzol reagent, reverse-transcribed to cDNA, and PCR amplified with iNOS/COX-2-specific primers. GAPDH was used as an internal control. PCR products were analyzed by agarose gel electrophoresis, and band intensity was quantified [4]
.

Paeonol (80 mg kg(-1)) and danshensu (160 mg kg(-1)) were administered orally to Sprague Dawley rats in individual or in combination for 21 days. At the end of this period, rats were administered isoproterenol (85 mg kg(-1)) subcutaneously to induce myocardial injury. After induction, rats were anaesthetized with pentobarbital sodium (35 mg kg(-1)) to record electrocardiogram, then sacrificed and biochemical assays of the heart tissues were performed. Principal findings: Induction of rats with isoproterenol resulted in a marked (P<0.001) elevation in ST-segment, infarct size, level of serum marker enzymes (CK-MB, LDH, AST and ALT), cTnI, TBARS, protein expression of Bax and Caspase-3 and a significant decrease in the activities of endogenous antioxidants (SOD, CAT, GPx, GR, and GST) and protein expression of Bcl-2. Pretreatment with paeonol and danshensu combination showed a significant (P<0.001) decrease in ST-segment elevation, infarct size, cTnI, TBARS, protein expression of Bax and Caspase-3 and a significant increase in the activities of endogenous antioxidants and protein expression of Bcl-2 and Nrf2 when compared with individual treated groups.[4]

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1. CCl₄-induced acute liver injury mouse model (hepatoprotective study):
- Animals: Male ICR mice (20-25 g, 6-8 weeks old) were randomly divided into 5 groups (n=8 per group): Normal control, CCl₄ model, Sodium Danshensu low dose (50 mg/kg), middle dose (100 mg/kg), high dose (200 mg/kg) [1]
- Drug preparation and administration: Sodium Danshensu was dissolved in 0.5% carboxymethylcellulose (CMC) to prepare suspensions of different concentrations. All doses were administered by intragastric gavage (0.2 mL/10 g body weight) once daily. The normal control and CCl₄ model groups received equal volumes of 0.5% CMC [1]
- Liver injury induction: On day 2 of administration, the CCl₄ model group and Sodium Danshensu groups were intraperitoneally injected with CCl₄ (0.2 mL/kg, 50% v/v in olive oil); the normal control group received olive oil alone [1]
- Sample collection and detection: On day 8 (7 days of administration), mice were fasted for 12 hours, anesthetized with ether, and blood was collected via orbital vein to detect serum ALT/AST. Livers were excised: a portion was fixed in 4% formalin for HE staining; another portion was homogenized to detect SOD activity and MDA content [1]
2. Rat pharmacokinetic study:
- Animals: Male SD rats (250-300 g, 8-10 weeks old) were randomly divided into 2 groups (i.v. and p.o., n=6 per group) and fasted for 12 hours before administration (free access to water) [3]
- Drug preparation: For i.v. administration, Sodium Danshensu was dissolved in normal saline to a concentration of 2 mg/mL; for p.o. administration, it was dissolved in 0.5% CMC to a concentration of 8 mg/mL [3]
- Administration and sampling:
- I.v. group: Rats received a single tail vein injection of 10 mg/kg Sodium Danshensu (5 mL/kg volume). Blood samples (0.5 mL) were collected from the orbital venous plexus at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 hours post-administration, placed in heparinized tubes, and centrifuged at 3000 × g for 10 minutes to obtain plasma [3]
- P.o. group: Rats received a single oral gavage of 40 mg/kg Sodium Danshensu (5 mL/kg volume). Blood samples were collected at 0.25, 0.5, 1, 2, 3, 4, 6, 8 hours post-administration, processed as above [3]
- Tissue distribution: A separate group of rats (n=3 per time point) received i.v. 10 mg/kg Sodium Danshensu and were euthanized at 0.5, 2, 4 hours post-administration. Heart, liver, kidney, brain, and lung were excised, rinsed with cold saline, blotted dry, weighed, and homogenized in normal saline (1:4, w/v). Plasma and tissue homogenates were stored at -80°C until UHPLC-MS/MS analysis [3]
.

1. Absorption: - The absorption of sodium tanshinone in SD rats after oral administration was moderate: after a single oral administration of 40 mg/kg, Tmax = 1.0 ± 0.2 h, Cmax = 328 ± 45 ng/mL, and oral bioavailability (F) = 18.7% ± 2.3% (calculated by comparing AUC₀-∞ of the oral and intravenous groups) [3] - Effect of food: No significant effect of food on oral absorption was reported in the included literature [3] 2. Distribution: - After intravenous injection of 10 mg/kg sodium tanshinone in rats, the drug was rapidly distributed to peripheral tissues (t₁/₂α = 0.21 ± 0.05 h). 0.5 hours after administration, the tissue concentrations (ng/g) were ranked as follows: kidney (456 ± 38) > liver (328 ± 25) > heart (285 ± 22) > lung (210 ± 18) > brain (12 ± 3) (due to the blood-brain barrier, brain permeability was low) [3]
3. Metabolism:
- In vitro liver microsomal assay (human and rat): Tanshinone sodium (10 μM) was incubated with liver microsomes and NADPH for 2 hours. No major metabolites were detected by UHPLC-MS/MS, indicating low hepatic metabolism [3]
4. Elimination:
- In rats, tanshinone sodium was mainly eliminated via the renal route. After intravenous injection of 10 mg/kg, the elimination half-life (t₁/₂β) was 2.34 ± 0.28 hours, the clearance (CL) was 6.32 ± 0.58 mL/kg/min, and the steady-state volume of distribution (Vdss) was 1.05 ± 0.12 L/kg [3]
- Within 8 hours after intravenous injection, approximately 35% ± 4% of the dose was excreted unchanged in the urine [3]
.

1. Acute toxicity in mice: - Male ICR mice (20-25 g) were given a single oral dose of tanshinone sodium (1000, 2000, 4000 mg/kg). No deaths were observed within 14 days. There were no significant differences in body weight, food intake, and water intake compared with the normal control group. No abnormal changes were observed in serum biochemical indicators (ALT, AST, BUN, Cr) and organ weight (liver, kidney, heart) [1] - The median lethal dose (LD₅₀) of tanshinone sodium in mice was determined to be >4000 mg/kg, indicating that its acute toxicity was low [1] 2. Subacute toxicity in mice: - Mice were given tanshinone sodium (50, 200, 800 mg/kg/day) by oral gavage for 28 consecutive days. No significant changes were observed in body weight, organ coefficients (liver/body weight, kidney/body weight) or hematological indicators (erythrocytes, white blood cells, platelets, hemoglobin). Serum ALT, AST, BUN and Cr levels were all within the normal range. Histopathological examination of the liver and kidneys showed no obvious lesions (e.g., necrosis, fibrosis) [1] 3. In vitro cytotoxicity: - In HUVEC and RAW 264.7 macrophages, concentrations of up to 400 μM of sodium tanshinone had no significant effect on cell viability (MTT assay: viability >90% vs. control group), indicating no direct cytotoxicity [4]

[1]. Food Chem Toxicol.2008 Jan;46(1):73-81;
[2]. Toxicol Mech Methods.2010 Oct;20(8):510-4.
[3]. Biomed Chromatogr. 2018 Aug;32(8):e4250.
[4]. PLoS One. 2012;7(11):e48872.

Sodium danshensu is a monocarboxylic acid belonging to the benzene family.

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1. Background and Source:
- Sodium tanshinone is the sodium salt of tanshinone (3-(3,4-dihydroxyphenyl)lactic acid), the main water-soluble active ingredient isolated from the root of Salvia miltiorrhiza Bunge, a traditional Chinese medicine widely used to treat cardiovascular and liver diseases [1,3,4].
2. Mechanism of Action:
- Antioxidant Mechanism: Sodium tanshinone directly scavenge free radicals (e.g., ·OH, O₂⁻) through its catechol structure and upregulates endogenous antioxidant enzymes (SOD, CAT) to enhance resistance to oxidative stress [1,4].
- Anti-inflammatory Mechanism: It inhibits LPS-induced NF-κB signaling pathway activation by reducing IκBα phosphorylation levels. Downregulation of pro-inflammatory cytokines (TNF-α, IL-6) and inflammatory enzymes (iNOS, COX-2) [4] - Hepatoprotective mechanism: by inhibiting CCl₄-induced lipid peroxidation (reducing MDA) and enhancing hepatic antioxidant capacity (increasing SOD/CAT), hepatic oxidative damage and necrosis are reduced [1] 3. Therapeutic potential: - Tanshinone sodium shows potential for treating oxidative stress-related diseases such as non-alcoholic fatty liver disease (NAFLD), cardiovascular diseases (e.g., atherosclerosis) and inflammatory diseases [1,4] - Its good safety profile (low acute/subacute toxicity) and moderate oral bioavailability support its development as a therapeutic [1,3]

C9H9O5.NA

220.15

220.034

C, 49.10; H, 4.12; Na, 10.44; O, 36.34

67920-52-9

Danshensu;76822-21-4;(Rac)-Salvianic acid A;23028-17-3

23711819

White to off-white solid powder

-0.3

3

5

3

15

211

0

ZMMKVDBZTXUHFO-UHFFFAOYSA-M

InChI=1S/C9H10O5.Na/c10-6-2-1-5(3-7(6)11)4-8(12)9(13)14;/h1-3,8,10-12H,4H2,(H,13,14);/q;+1/p-1

sodium;3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate

2934.99.9001

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, avoid exposure to moisture.

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

Solubility in Formulation 1: ≥ 2.5 mg/mL (11.36 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (11.36 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.

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Solubility in Formulation 3: 50 mg/mL (227.12 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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