The TGF-β/Smad signaling pathway. Scoparone downregulates the expression of TGF-β and p-Smad2/3, and upregulates the expression of Smad7. [1]

The maximum inhibitory effect of scopone (0-0.4 nM; 24-48 hours) on PSC proliferation is observed at 0.4 nM [1].

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Scoparone significantly inhibited the proliferation of pancreatic stellate cells (PSCs) in a dose-dependent manner. At 24 and 48 hours post-treatment, the high concentration (0.4 mmol/L) produced the most significant inhibition compared to the control group (P < 0.01). [1]
- Scoparone reduced oxidative stress in PSCs. Treatment with different doses (0.1, 0.2, 0.4 mmol/L) significantly decreased intracellular malondialdehyde (MDA) levels and increased superoxide dismutase (SOD) levels at 48 hours compared to the control group (P < 0.01). The high concentration (0.4 mmol/L) resulted in the lowest MDA and highest SOD levels. [1]
- Scoparone inhibited epithelial-mesenchymal transition (EMT) and fibrosis in PSCs. Immunofluorescence and Western blot analysis showed that treatment with different doses (0.1, 0.2, 0.4 mmol/L) significantly reduced the expression of mesenchymal and fibrotic markers (α-SMA, Collagen I, Vimentin) and increased the expression of the epithelial marker E-cadherin compared to the control group. [1]
- Scoparone modulated the TGF-β/Smad pathway in PSCs. Western blot analysis revealed that treatment with different doses (0.1, 0.2, 0.4 mmol/L) decreased the expression of TGF-β and p-Smad2/3, while increasing the expression of Smad7 compared to the control group. [1]

Rats with DBTC-induced pancreatitis benefit greatly from scopone (oral; 30 mg/kg, 60 mg/kg; 4 weeks) in terms of pancreatic weight loss [1].

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In a rat model of chronic pancreatitis induced by dibutyltin dichloride (DBTC), oral administration of Scoparone (30 mg/kg and 60 mg/kg) significantly ameliorated pancreatic damage. This was evidenced by a significant increase in the pancreas wet weight/body weight ratio and significant attenuation of histological markers of damage (inflammation, edema, abnormal architecture, glandular atrophy, fibrosis, and pseudotubular complexes) compared to the untreated chronic pancreatitis (CP) group (P < 0.05). [1]
- Scoparone reduced oxidative stress in pancreatic tissues. Oral administration of Scoparone (30 mg/kg and 60 mg/kg) significantly decreased MDA levels and increased SOD levels in the pancreas of DBTC-induced chronic pancreatitis rats compared to the untreated CP group (P < 0.05). The high dose (60 mg/kg) caused significant decreases in MDA and increases in SOD compared to the low dose (30 mg/kg) (P < 0.05). [1]
- Scoparone inhibited PSC activation and the TGF-β/Smad signaling pathway in the pancreas. Western blot analysis showed that oral administration of Scoparone (especially at 60 mg/kg) significantly reduced the expression of α-SMA and collagen I, decreased TGF-β and p-Smad2/3 levels, and increased Smad7 expression in the pancreas of DBTC-induced chronic pancreatitis rats compared to the untreated CP group. [1]

Isolation and Culture of Pancreatic Stellate Cells (PSCs): PSCs were isolated from rat pancreas tissue blocks and cultured in DMEM/F12 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C with 5% CO2. Cells used for experiments were between passages 3-5. Identification was performed by immunofluorescence using anti-desmin, anti-GFAP, and anti-α-SMA antibodies. [1]
- MTT Assay for Cell Proliferation: PSCs were seeded in 96-well plates at a density of 5 × 10³ cells per well. After 24 and 48 hours of incubation with different concentrations of Scoparone, 20 µL of MTT solution (5 mg/mL) was added to each well and incubated for 4 hours. The MTT solution was then removed, and 200 µL of DMSO was added to dissolve the formazan crystals for 15 minutes. The optical density (OD) was measured at 490 nm using a microplate reader. [1]
- Measurement of MDA and SOD Levels: PSCs were seeded in 6-well plates at a density of 2 × 10⁴ cells per well. After 48 hours of treatment, the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the cells were detected using commercially available kits, following the manufacturer's instructions. [1]
- Immunofluorescence Staining: PSCs were seeded in 24-well plates at a density of 1 × 10⁴ cells per well. After treatment, cells were stained with primary antibodies against α-SMA, vimentin, collagen I, and E-cadherin. Hoechst 33342 was used for nuclear counterstaining. Staining was observed using confocal microscopy. [1]
- Western Blotting: Total protein from PSCs was extracted, quantified using the BCA method, separated by SDS-PAGE, and transferred to membranes. Membranes were incubated with primary antibodies against α-SMA, collagen I, E-cadherin, vimentin, TGF-β, Smad2/3, p-Smad2/3, and Smad7, followed by secondary antibodies. Protein bands were visualized using an ECL kit and quantified with a gel imaging system. [1]

Animal/Disease Models: Chronic pancreatitis (CP) rat model [1]
Doses: 30 mg/kg, 60 mg/kg
Route of Administration: po (po (oral gavage)) 30 mg/kg, 60 mg/kg; 4-week
Experimental Results: Prevent pancreatic damage

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Chronic Pancreatitis (CP) Rat Model and Treatment:** Male Sprague Dawley rats (250-300g) were used. Chronic pancreatitis was induced by a single caudal vein injection of dibutyltin dichloride (DBTC) at a dose of 8 mg/kg. DBTC was dissolved in a mixture of ethanol and glycerol (2:3 ratio) to a final concentration of 8 mg/mL. The control group received the vehicle (ethanol/glycerol mixture). One day after DBTC infusion, rats were randomly assigned to treatment groups. Scoparone was administered orally at doses of 30 mg/kg (low dose) or 60 mg/kg (high dose) in 1.5 mL of saline daily. The salvianolic acid B group received 10 mg/kg orally. The control and CP groups received the same volume of saline. The treatment continued for 4 weeks, after which pancreatic tissue and serum were collected for analysis. [1]
- **Histological Examination:** After sacrifice, pancreases were removed, weighed, and processed for histological analysis. Two pathologists scored the tissue samples on six indicators: inflammation, edema, abnormal architecture, glandular atrophy, fibrosis, and pseudotubular complexes. [1]
- **Biochemical Parameter Detection:** Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine (Cr) were assayed using commercially available kits to assess liver and kidney function. [1]
- **Measurement of Pancreatic MDA and SOD:** Pancreatic tissue homogenates were used to quantify the levels of MDA and SOD using commercially available detection kits, following the manufacturer's instructions. [1]
- **Western Blotting of Pancreatic Tissue:** The expression of α-SMA, collagen I, TGF-β, Smad2/3, p-Smad2/3, and Smad7 in pancreatic tissue was determined by Western blotting, following similar procedures as described for the cell assays. [1]

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Chronic Pancreatitis (CP) Rat Model and Treatment: Male Sprague Dawley rats (250-300g) were used. Chronic pancreatitis was induced by a single caudal vein injection of dibutyltin dichloride (DBTC) at a dose of 8 mg/kg. DBTC was dissolved in a mixture of ethanol and glycerol (2:3 ratio) to a final concentration of 8 mg/mL. The control group received the vehicle (ethanol/glycerol mixture). One day after DBTC infusion, rats were randomly assigned to treatment groups. Scoparone was administered orally at doses of 30 mg/kg (low dose) or 60 mg/kg (high dose) in 1.5 mL of saline daily. The salvianolic acid B group received 10 mg/kg orally. The control and CP groups received the same volume of saline. The treatment continued for 4 weeks, after which pancreatic tissue and serum were collected for analysis. [1]
- Histological Examination: After sacrifice, pancreases were removed, weighed, and processed for histological analysis. Two pathologists scored the tissue samples on six indicators: inflammation, edema, abnormal architecture, glandular atrophy, fibrosis, and pseudotubular complexes. [1]
- Biochemical Parameter Detection: Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine (Cr) were assayed using commercially available kits to assess liver and kidney function. [1]
- Measurement of Pancreatic MDA and SOD: Pancreatic tissue homogenates were used to quantify the levels of MDA and SOD using commercially available detection kits, following the manufacturer's instructions. [1]
- Western Blotting of Pancreatic Tissue: The expression of α-SMA, collagen I, TGF-β, Smad2/3, p-Smad2/3, and Smad7 in pancreatic tissue was determined by Western blotting, following similar procedures as described for the cell assays. [1]

Scoparone showed no significant negative impact on liver and kidney function. Serum levels of AST, ALT, BUN, and Cr showed no significant differences between the control, low-dose Scoparone (30 mg/kg), and high-dose Scoparone (60 mg/kg) treatment groups in the rat model (P > 0.05). [1]

[1]. Scoparone Protects Against Pancreatic Fibrosis via TGF-β/Smad Signaling in Rats. Cell Physiol Biochem. 2016;40(1-2):277-286. Epub 2016 Nov 18.

Scoparone belongs to the coumarin class of compounds, and its structure is similar to that of aescin, except that the two hydroxyl groups at positions 6 and 7 are replaced by methoxy groups. It is a major component of the traditional Chinese medicine anemone and possesses various pharmacological activities, such as anti-inflammatory, anti-allergic, and anti-tumor activities. It can be used as a plant metabolite, anti-inflammatory drug, lipid-lowering drug, immunosuppressant, antihypertensive drug, and anti-allergic drug. Scoparone belongs to the coumarin class of aromatic ethers, and its function is similar to that of aescin. Scoparone has been reported to exist in dwarf snow lotus (Saussurea eopygmaea), Cedrelopsis grevei, and other organisms with relevant data.

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Scoparone is the main component of the traditional Chinese herb Artemisia capillaris (Yin Chen Hao). It is known for its antioxidant and anti-inflammatory properties. [1]
- In this study, Scoparone was shown to protect against pancreatic fibrosis by reducing oxidative stress, repressing pancreatic stellate cell activation, and inhibiting the epithelial-mesenchymal transition (EMT) process, primarily through the regulation of the TGF-β/Smad signaling pathway. [1]

C11H10O4

206.1947

206.057

C, 64.08; H, 4.89; O, 31.04

120-08-1

8417

Solid powder

1.2±0.1 g/cm3

369.2±42.0 °C at 760 mmHg

145°C

166.8±27.9 °C

0.0±0.8 mmHg at 25°C

1.557

1.6

0

4

2

15

274

0

O1C(C([H])=C([H])C2=C([H])C(=C(C([H])=C12)OC([H])([H])[H])OC([H])([H])[H])=O

GUAFOGOEJLSQBT-UHFFFAOYSA-N

InChI=1S/C11H10O4/c1-13-9-5-7-3-4-11(12)15-8(7)6-10(9)14-2/h3-6H,1-2H3

6,7-dimethoxychromen-2-one

Escoparone; Scoparone

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 : ~50 mg/mL (~242.49 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (10.09 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (10.09 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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (10.09 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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