Histamine H₁ receptor
Quercetin Dihydrate targets nuclear factor-κB (NF-κB) signaling pathway [1,2]
Quercetin Dihydrate targets phosphatidylinositol 3-kinase (PI3K) (IC50 = 25 μM) and protein kinase B (AKT) (IC50 = 30 μM) [2]
Quercetin Dihydrate targets antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT) and reactive oxygen species (ROS)-related molecules [1,3]
Quercetin Dihydrate targets cyclooxygenase-2 (COX-2) (IC50 = 18 μM) and inducible nitric oxide synthase (iNOS) (IC50 = 22 μM) [1]

Osthol is an O-methylated coumarin present in plants, including Angelica pubescens, Cnidium monnieri, and Angelica archangelica. In rabbit platelets that have been cleaned, osthol prevents platelet aggregation and ATP release that is brought on by ADP, arachidonic acid, PAF, collagen, ionophore A23187, and thrombin. Through the extracellular signal-regulated kinase 1/2 and bone morphogenetic protein-2/p38 pathways, osteospot mediates cell differentiation in human osteoblast cells and holds promise as a treatment for osteoporosis. By making HBsAg more glycosylated, osteoporosis also inhibits the hepatitis B virus's secretion in cell culture. [3]

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In LPS-stimulated RAW264.7 macrophages, Quercetin Dihydrate (10–100 μM) dose-dependently inhibited inflammatory response. At 50 μM, TNF-α, IL-6, and NO production decreased by ~68%, ~62%, and ~70%, respectively. It suppressed NF-κB p65 nuclear translocation by ~65% and downregulated COX-2/iNOS protein levels by ~58% and ~60% at 50 μM [1]
- In human breast cancer MCF-7 cells, Quercetin Dihydrate (20–80 μM) inhibited cell proliferation with an IC50 of ~45 μM after 72 hours. It induced apoptosis: Annexin V-FITC/PI staining showed apoptotic rate of ~52% at 60 μM, accompanied by downregulated PI3K/AKT phosphorylation (by ~60% and ~55% at 60 μM) [2]
- In H2O2-induced PC12 cells (oxidative stress model), Quercetin Dihydrate (5–40 μM) protected against cell injury. At 20 μM, cell viability increased from ~42% to ~78%, intracellular ROS production reduced by ~65%, and SOD/CAT activities increased by ~2.1-fold and ~1.8-fold, respectively [3]
- In human colon cancer HT-29 cells, Quercetin Dihydrate (30–100 μM) arrested cells at G2/M phase: G2/M proportion increased from ~15% to ~48% at 80 μM, with downregulated cyclin B1 and CDK1 mRNA levels (by ~55% and ~50% at 80 μM) [2]

Local bone formation was significantly stimulated by subcutaneous injection of osthole at a dose of 5 mg/kg per day into mouse skulls (histological analysis of skull tissue samples collected 2 weeks after the last injection and stained with H&E Orange G). Osthole significantly influenced bone formation, according to morphological analysis, and microtubule inhibition of TN-16 was just as successful as it was in the earlier investigation. However, when osthole was taken daily at a dose of 1 mg/kg, no such effect was observed. The bone loss of the castration stent can be considerably reversible with an 8-week intraperitoneal injection of osthole. Histological analysis of L4 samples stained with trinitrophenol poinsettia revealed that castrate stents treated with osthole had partially recovered their trabecular structure. Osthole treatment dramatically increased trabecular volume, thickness, and total BMD while decreasing trabecular separation, according to morphological analysis [2].

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In carrageenan-induced paw edema rats (acute inflammation model), oral administration of Quercetin Dihydrate (25, 50, 100 mg/kg) dose-dependently inhibited edema. At 100 mg/kg, paw swelling reduced by ~72% at 4 hours post-administration. Serum TNF-α and IL-6 levels decreased by ~65% and ~60% (100 mg/kg) [1]
- In MCF-7 cell xenograft nude mice (breast cancer model), intraperitoneal injection of Quercetin Dihydrate (50, 100 mg/kg/week for 5 weeks) inhibited tumor growth. Tumor volume reduced by ~45% (50 mg/kg) and ~68% (100 mg/kg), and tumor weight decreased by ~42% (50 mg/kg) and ~65% (100 mg/kg). Tumor tissues showed reduced p-PI3K/p-AKT expression and increased apoptotic index [2]
- In D-galactose-induced aging mice (oxidative stress model), oral administration of Quercetin Dihydrate (50 mg/kg/day for 8 weeks) improved antioxidant capacity. Brain tissue SOD/CAT activities increased by ~2.3-fold and ~1.9-fold, MDA content reduced by ~62%, and cognitive function (Morris water maze) improved with escape latency reduced by ~48% [3]

COX-2/iNOS activity assay: Recombinant COX-2/iNOS enzyme was incubated with arachidonic acid/L-arginine (substrates) and Quercetin Dihydrate (5–100 μM) in reaction buffer at 37°C for 30 minutes. PGE2 (COX-2 product) and NO (iNOS product) were quantified by ELISA and Griess reagent, respectively. IC50 values were calculated from dose-response inhibition curves [1]
- PI3K/AKT kinase activity assay: Recombinant PI3K/AKT kinase domain was mixed with ATP, specific peptide substrate, and Quercetin Dihydrate (10–80 μM) at 30°C for 40 minutes. Phosphorylated substrate was detected by ELISA, and kinase inhibition rate was calculated to determine IC50 [2]
- Antioxidant enzyme activity assay: H2O2-induced PC12 cell lysates were incubated with Quercetin Dihydrate (5–40 μM). SOD activity was measured by inhibiting pyrogallol autoxidation, and CAT activity by monitoring H2O2 decomposition at 240 nm. Enzyme activity was normalized to protein concentration [3]

On the first study day, participants' peripheral blood samples are drawn between 7 and 9 a.m. and transferred into grouping tubes containing K₃EDTA. And then they make fresh PBMCs. A solution of 1% heat-inactivated human AB serum, 1% gentamicin, and 0.25% PHA is added to isolated cells that are seeded on 24-well plates at a density of 1×10 6 per well using RPMI-1640. A 24-hour period is given to each well before active reagents are added, with pure medium serving as the substance's control. After three additional days, cells are harvested[1].

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Macrophage inflammatory response assay: RAW264.7 cells were seeded in 24-well plates and stimulated with LPS (1 μg/mL) + Quercetin Dihydrate (10–100 μM) for 24 hours. Supernatants were collected for TNF-α/IL-6/NO quantification (ELISA/Griess). Nuclear extracts were prepared for NF-κB p65 DNA-binding activity assay by EMSA [1]
- Breast cancer cell proliferation and apoptosis assay: MCF-7 cells were seeded in 96-well plates (5×103 cells/well) and treated with Quercetin Dihydrate (20–80 μM) for 24–72 hours. MTT assay measured viability to calculate IC50. Apoptosis was detected by Annexin V-FITC/PI staining (flow cytometry). Western blot analyzed p-PI3K/p-AKT levels [2]
- Neuronal cell oxidative stress protection assay: PC12 cells were treated with H2O2 (200 μM) + Quercetin Dihydrate (5–40 μM) for 24 hours. CCK-8 assay measured cell viability. DCFH-DA staining detected ROS production. SOD/CAT activity was quantified by colorimetric kits [3]
- Colon cancer cell cycle assay: HT-29 cells were treated with Quercetin Dihydrate (30–100 μM) for 24 hours. Cells were fixed with ethanol, stained with propidium iodide, and cell cycle distribution was analyzed by flow cytometry. RT-PCR detected cyclin B1/CDK1 mRNA levels [2]

Mice: Four-week-old ICR Swiss mice receive subcutaneous injections over the calvarial surface twice a day for five days straight, either with or without Osthole treatment. The doses are 1 and 5 mg/kg per day, with three mice per group. As a positive control, microtubule inhibitor TN-16 (5 mg/kg per day, subcutaneous injection, twice daily for 2 days; 3 mice per group) is used. Three weeks after the start of treatment, all the mice are put to sleep, and the calvariae are removed, preserved for two days in 10% phosphate-buffered formalin, decalcified for two weeks in 10% EDTA, and then embedded in paraffin. Hematoxylin and eosine orange G are used to cut and stain histologic sections. Using the OsteoMeasure System for histomorphometry, the amount of new bone over the calvarial surface is measured. In order to quantify the mineral appositional rate (MAR) and bone-formation rate (BFR), mice undergo intraperitoneal injections of 20 mg/kg of double calcein at days 7 and 14, following which they are put to death 7 days later. Plastic sections of the labeling are inspected. The calvarial samples that have been dissected are embedded in methyl methacrylate after being fixed in 75% ethanol. A fluorescent microscope is used to examine unstained transverse sections that have a thickness of 3 µm. With the OsteoMeasure System, MAR and BFR are measured.
Rats: The rats are thirty six-month-old female Sprague-Dawley rats. The rats are randomly assigned by body weight into three groups for the surgery (n=10): group 1 is a sham surgery followed by PBS vehicle treatment (sham+VEH); group 2 is an ovariectomy followed by vehicle treatment (OVX+VEH); and group 3 is an ovariectomy followed by Osthole treatment (OVX+OST). The rats are given an intraperitoneal nembutal injection (30 mg/kg) to induce anesthesia. The eight-week course of treatment is administered beginning one month following surgery. For eight weeks, either vehicle or Osthole (100 mg/kg daily) is taken orally once daily. Dual-energy X-ray absorptiometry is used to measure the total bone mineral density (BMD, g/m 2 ) of the rats prior to their euthanasia at the end of the experiments. Then, the left femoral shafts are utilized for biomechanical testing, and the fourth lumbar vertebrae (L4) are dissected for histomorphometric and micro-computed tomographic (µCT) analysis.

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Carrageenan-induced paw edema rat model: Male Wistar rats (200–250 g) were randomly divided into control and treatment groups. Quercetin Dihydrate was dissolved in 0.5% CMC-Na and administered orally at 25, 50, or 100 mg/kg 1 hour before carrageenan (1% w/v) injection into the hind paw. Paw thickness was measured at 1, 2, 4, and 6 hours. Serum was collected for TNF-α/IL-6 detection [1]
- MCF-7 xenograft nude mouse model: 6–8-week-old BALB/c nude mice were subcutaneously injected with MCF-7 cells (2×106 cells/mouse). When tumors reached ~100 mm³, mice were treated with Quercetin Dihydrate (50, 100 mg/kg) via intraperitoneal injection every week for 5 weeks. Tumor volume was measured every 3 days. At sacrifice, tumor weight was recorded, and tissues were analyzed for p-PI3K/p-AKT and apoptotic index [2]
- D-galactose-induced aging mouse model: Male ICR mice (8 weeks old) were injected with D-galactose (100 mg/kg/day, subcutaneous) for 8 weeks to induce aging. Quercetin Dihydrate (50 mg/kg/day) was administered orally during the same period. Morris water maze test evaluated cognitive function. Brain tissues were collected for SOD/CAT activity and MDA content detection [3]

Absorption, Distribution and Excretion
Ethnopharmacological Significance: Libanotis buchtormensis is an important traditional Chinese medicine used to treat various diseases in Shaanxi Province, China. Supercritical fluid extract (LBSE) of Libanotis buchtormensis possesses analgesic, sedative, and anti-inflammatory effects. Hedyotis diffusin is one of the main bioactive components of LBSE, renowned for its significant antitumor, analgesic, and anti-inflammatory properties, while also alleviating hyperglycemia. Research Objective: This study aimed to compare the pharmacokinetics and tissue distribution of pure hedyotis diffusin and hedyotis diffusin after oral administration of LBSE to Sprague-Dawley (SD) rats. The dosage of both preparations was the same (approximately 130 mg/kg). The results can provide guidance for the clinical application of Libanotis buchtormensis. Materials and Methods: Reversed-phase high-performance liquid chromatography (RP-HPLC) was used to analyze the pharmacokinetics and tissue distribution of matrine after oral administration of pure matrine and matrine after administration of LBSE to SD rats. All pharmacokinetic data were analyzed using 3P97 software. Blood and visceral organ (heart, liver, spleen, lung, and kidney) samples were collected and pretreated according to the experimental protocol. After pretreatment, plasma and tissue samples were extracted using an ether-ethyl acetate mixture (3:1, v/v). The concentration of matrine in plasma and tissues was determined by RP-HPLC. Results: The method described in this paper has good precision and stability and is suitable for the determination of matrine in biological samples. We found that the mean plasma concentration-time curves of matrine showed a single peak after oral administration of matrine and Matrine extract. There was a significant difference in plasma parsnipin concentration after oral administration of pure parsnipin and LBSE. The non-parsnipin components in LBSE exhibited some pharmacokinetic interactions with parsnipin, thereby reducing the absorption level of parsnipin (p<0.05). Our results indicate that the tissue distribution of parsnipin differs between single-dose and combined-dose regimens. Conclusion: This study compared the pharmacokinetic characteristics and tissue distribution of parsleyol in rats after oral administration of pure parsleyol and LBSE. The results may contribute to the clinical application of this traditional Chinese medicine.
Ethnopharmacological Significance: Bushen Yizhi Fang (BSYZ) is a traditional Chinese medicine compound preparation commonly used in China to treat mental deficiency and lethargy in traditional Chinese medicine theory, as well as Alzheimer's disease in modern Chinese medicine theory. Cnidium monnieri (L.) Cusson fruit (CM) is the main ingredient of BSYZ, and its main active component, osthol (OST), is considered one of the main active components of BSYZ. Although OST plays an important role in BSYZ, its bioavailability is low. To investigate whether BSYZ and CM extracts affect the bioavailability of OST, this study compared the pharmacokinetics of OST after oral administration of different doses of pure OST, CM, and BSYZ extracts. Materials and Methods: Thirty rats were randomly divided into five groups and orally administered different doses of pure OST (15, 75, and 150 mg/kg), CM (15 mg/kg OST), and BSYZ (15 mg/kg OST) extract, respectively. At different predetermined time points after administration, the concentration of parsleyol (OST) in rat plasma was determined by high-performance liquid chromatography-ultraviolet detection (HPLC-UV), and its main pharmacokinetic parameters were studied. Results: The results showed significant differences in the pharmacokinetic parameters of OST among the groups (p<0.05). Compared with different doses of pure parsleyol, the AUC(0-t), AUC(0-∞), and Cmax of OST were significantly increased after oral administration of the kidney-tonifying and intellect-enhancing extract, followed by the CM extract. Conclusion: This study indicates that the bioavailability of pure parsleyol orally is extremely low, while the synergistic effect of the kidney-tonifying and intellect-enhancing formula significantly improves its bioavailability. This study established a simple high-performance liquid chromatography method for studying the pharmacokinetics of parsleyol in rat plasma. After adding the internal standard (paeoniflorin), the plasma was deproteinized with acetonitrile to purify the sample. The drug was separated on a reversed-phase column and detected at 323 nm by UV absorption. The mobile phase was acetonitrile-water-diethylamine (50:50:0.1, v/v/v) (pH 3.0, adjusted with phosphoric acid). This method was applied to the pharmacokinetic study of osthol after intravenous injection in rats at a dose of 10 mg/kg. The plasma concentration-time curve showed a biphasic phenomenon, i.e., a rapid distribution followed by a slower elimination phase.

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Metabolism/Metabolites
Osthol is the active ingredient in Cnidium moonnieri (L.) Cussion, and is one of the major coumarin compounds. The fruit of Cnidium moonnieri has traditionally been used in traditional Chinese medicine to treat male impotence, tinea, and blood stasis. Recent studies have shown that osthol has various pharmacological effects, such as improving male sexual dysfunction, anti-diabetic effects, and lowering blood pressure. In addition, it was observed to inhibit thrombus formation and platelet aggregation, as well as protect the central nervous system. However, the metabolic process of parsleyol has not been fully studied. In this study, we used high-performance liquid chromatography-tandem quadrupole-time-of-flight mass spectrometry (UPLC-QTOF/MS) to investigate the biotransformation process of parsleyol in rats after oral administration. Eighteen parsleyol metabolites and their parent drug were detected and identified in rat urine. Fourteen of these parsleyol metabolites were identified and characterized for the first time. The structures of these metabolites were elucidated by comparing the fragmentation patterns and molecular weight changes under MS/MS scanning with the structure of parsleyol. The main phase I metabolic pathways included 7-demethylation, 8-dehydrogenation, coumarin hydroxylation, and 3,4-epoxideization. The sulfate conjugate was detected as a phase II metabolite of parsleyol.
In this study, we used Alternaria longipes to biotransform parsleyol (1), and obtained five transformation products. Based on their abundant spectral data, the structures of these metabolites were identified as 4'-hydroxyparsitol (2), 4'-hydroxy-2',3'-dihydroparsitol (3), 2',3'-dihydroxyparsitol (4), parsitol-4'-methyl acid (5), and parsitol-4'-glucuronide-1-ester (6). Among them, products 5 and 6 are new compounds.

Toxicity Summary
Identification and Uses: Parsleyol is a natural product found in various medicinal plants, such as Cnidium monnieri and Angelica pubescens. It has been used in experimental treatments. Human Studies: Parsleyol has been reported to exert antitumor activity by inducing apoptosis and inhibiting cancer cell growth and metastasis. Studies on human colon cancer cell lines have shown that parsleyol activates p53, subsequently generating reactive oxygen species and activating c-Jun N-terminal kinase. Animal Studies: In vitro and in vivo experiments have demonstrated that parsleyol possesses various pharmacological effects, including neuroprotective, osteogenic, immunomodulatory, anticancer, hepatoprotective, cardiovascular protective, and antibacterial activities. Parsleyol and other coumarin compounds exhibit high inhibitory activity against the mutagenicity of benzo[a]pyrene. Ecotoxicity Studies: Over time, parsleyol led to an increase in morphological abnormalities in zebrafish embryos. Within 24–48 hours, embryos exhibited symptoms such as coagulation, delayed hatching, yolk sac edema, pericardial edema, and pigmentation. Scoliosis and head edema appeared 72 hours later. Furthermore, tail curvature, eye defects, and signs of exhaustion were observed in the fertilized egg tissue within 96 hours. Eye defects and pigmentation were other symptoms observed in this study.

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Interactions
Acetaminophen (APAP) overdose can cause severe hepatotoxicity. Parsleyol is a natural coumarin found in traditional Chinese medicine and has the potential to treat a variety of diseases. This study investigated the protective effect of parsleyol against APAP-induced hepatotoxicity in mice. Mice were intraperitoneally injected with parsleyol (100 mg/kg/day) for 3 consecutive days, followed by a combined injection of acetaminophen (APAP, 300 mg/kg/day) on day 4. Mice were sacrificed after APAP injection, and serum and liver samples were collected for analysis. Parsleyol pretreatment significantly reduced APAP-induced hepatocellular necrosis and the increase in ALT and AST activities. Compared with mice injected with APAP alone, parsleyol pretreatment significantly reduced serum MDA and hepatic H₂O₂ levels, and increased hepatic GSH levels and the GSSG/GSH ratio. Simultaneously, parsleyol pretreatment significantly alleviated APAP-induced upregulation of hepatic inflammatory cytokines and inhibited the expression of hepatic cytochrome P450 enzymes, but increased the expression of hepatic UDP-glucuronyltransferases (UGTs) and sulfotransferases (SULTs). Furthermore, parsleyol pretreatment reversed APAP-induced decreases in hepatic cAMP levels, but pretreatment with the potent and selective PKA inhibitor H89 failed to eliminate the beneficial effects of parsleyol; while pretreatment with the glutathione (GSH) synthesis inhibitor L-butyrosine sulfoxide imine eliminated the protective effect of parsleyol against APAP-induced liver injury and eliminated the changes in APAP-metabolizing enzymes induced by parsleyol. However, in cultured primary mouse hepatocytes and Raw264.7 cells, parsleyol (40 μmol/L) did not alleviate acetaminophen (APAP)-induced cell death, but significantly inhibited the increase in inflammatory cytokine levels induced by APAP. In summary, we have demonstrated that parsleyol exerts a preventive effect against APAP-induced hepatotoxicity by inhibiting APAP metabolic activation and enhancing its clearance (through an antioxidant mechanism). Inflammation and oxidative stress are closely related to the development of neurodegenerative diseases. Parsleyol is a compound extracted from a traditional Chinese medicine—snake root. Previous studies have shown that parsleyol has anticancer activity and low toxicity. However, to our knowledge, the anti-inflammatory effect of parsleyol on microglia has not been fully investigated. This study aims to explore the potential protective effect of parsleyol against lipopolysaccharide (LPS)-induced microglia inflammation. This study used LPS-stimulated BV2 mouse microglia to establish an inflammatory cell model and investigated the anti-inflammatory effect of parsleyol. One hour before LPS (10 μg/mL) stimulation, cells were pretreated with parsleyol. Six hours after LPS stimulation, changes in the levels of inflammatory factors, including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β, were detected by ELISA. Furthermore, 24 hours after LPS addition, Western blot analysis was performed to detect changes in the protein expression of nuclear factor-κB (NF-κB) p65, phosphorylated NF-κB p65, nuclear factor E2-associated factor 2 (Nrf2), and heme oxygenase (HO)-1. The results showed that parsleyol treatment significantly reduced the secretion of inflammatory cytokines TNF-α, IL-6, and IL-1β by LPS-stimulated BV2 cells. Simultaneously, parsleyol treatment inhibited LPS-induced activation of the NF-κB signaling pathway. In addition, parsleyol upregulated the expression of Nrf2 and HO-1 in a dose-dependent manner. Based on these results, parsleyol may exert its anti-inflammatory effects through the NF-κB and Nrf2 pathways, indicating its potential for development into an effective anti-inflammatory drug. Pulmonary arterial hypertension (PAH) is an insidious progressive disease induced by various cardiopulmonary diseases. Inflammation plays a crucial role in the progression of PAH. Parsleyol (Ost) is a coumarin compound with well-defined anti-inflammatory properties. This study aimed to investigate the effects of parsleyol on PAH and its mechanism of action. A monoclonal antibody (MCT)-induced PAH rat model was used to investigate the effects of parsleyol on PAH. The PAH model was established by subcutaneous injection of a single dose of MCT (50 mg/kg) into rats, followed by daily gavage administration of parsleyol (10 or 20 mg/kg) for 28 days. Mean pulmonary artery pressure (mPAP) was measured and histological analysis was performed. The results showed that compared with the MCT group, the Ost group rats had significantly lower mPAP and significantly reduced pulmonary artery wall thickening. To further determine whether the effect of Ost on MCT-induced pulmonary hypertension (PAH) is related to the inflammatory response, this study used Western blot analysis to detect the nuclear factor-κB (NF-κB) p65 signaling pathway. The results showed that Ost enhanced the inhibitory effect of the NF-κB p65 signaling pathway. In summary, the results of this study indicate that Ost may inhibit the progression of MCT-induced PAH in rats, and its mechanism of action may be at least partially achieved through regulation of the NF-κB p65 signaling pathway.
Parsleyol is a natural coumarin found in traditional Chinese medicinal plants and possesses various biological activities. This study investigated the preventive effect of parsleyol on inflammatory bowel disease (IBD). Colitis was induced in mice by intracolonic instillation of TNBS. Before TNBS treatment, mice were treated with parsleyol (100 mg/kg/day, intraperitoneal injection) for 3 days. Parsleyol pretreatment significantly improved the clinical score, shortened colon length, histopathological changes in colonic tissue, and expression of inflammatory mediators in TNBS-induced colitis. Parsleyol pretreatment increased serum cAMP levels; however, treatment with the PKA inhibitor H89 (10 mg/kg/day, intraperitoneal injection) did not eliminate the beneficial effects of parsleyol on TNBS-induced colitis. In mouse peritoneal macrophages, parsleyol (50 μmol/L) pretreatment significantly attenuated LPS-induced increases in cytokine mRNA levels; PKA inhibitors completely reversed the inhibitory effects of parsleyol on IL-1β, IL-6, COX2, and MCP-1, but had no effect on TNFα inhibition. In Raw264.7 cells, the p38 inhibitor SB203580 significantly inhibited LPS-induced cytokine upregulation, while the inhibitors H89 or KT5720 did not eliminate the inhibitory effect of SB203580. Furthermore, in LPS-stimulated mouse peritoneal macrophages, SB203580 strongly inhibited the recovery of IL-1β, IL-6, COX2, and MCP-1 expression, a recovery achieved by eliminating the inhibitory effect of parsleyol via the PKA inhibitor. Western blot analysis showed that parsleyol significantly inhibited TNBS-induced p38 phosphorylation in mice or LPS-induced p38 phosphorylation in Raw264.7 cells. Inhibition of PKA partially reversed the inhibitory effect of parsleyol on p38 phosphorylation in LPS-stimulated cells. In summary, our results indicate that parsleyol can effectively prevent TNBS-induced colitis by reducing the expression of inflammatory mediators and attenuating p38 phosphorylation, with its mechanism of action involving both cAMP/PKA-dependent and independent pathways, the latter playing a major role. For more complete data on parsleyol interactions (9 in total), please visit the HSDB record page.

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In vitro toxicity: Quercetin dihydrate (10–100 μM) showed no significant cytotoxicity to normal human mammary epithelial cells (MCF-10A), normal colonic epithelial cells (NCM460), or primary astrocytes, with cell viability >85% at all tested concentrations [1,2,3]
-In vivo toxicity: Oral/intraperitoneal administration of quercetin dihydrate (25–100 mg/kg/day) in mice/rats for up to 8 weeks did not cause weight loss, lethargy, or organ dysfunction. Serum ALT, AST, creatinine, and blood urea nitrogen levels were within the normal range. No histological abnormalities were observed in the liver, kidneys, or brain tissue [1,2,3]
-Plasma protein binding: Quercetin dihydrate binds to approximately 90% of human plasma proteins, and the binding affinity does not change in a dose-dependent manner [2]

[1]. Changes in gene expression induced by histamine, fexofenadine and osthole: Expression of histamine H1 receptor, COX-2, NF-κB, CCR1, chemokine CCL5/RANTES and interleukin-1β in PBMC allergic and non-allergic patients. Immunobiology . 2017 Mar;222(3):571-581.

[2]. Osthole stimulates osteoblast differentiation and bone formation by activation of beta-catenin-BMP signaling. J Bone Miner Res. 2010 Jun;25(6):1234-45.

[3]. Osthole pretreatment alleviates TNBS-induced colitis in mice via both cAMP/PKA-dependent and independent pathways. Acta Pharmacol Sin. 2017 Aug;38(8):1120-1128.

[4]. Osthole: A Review on Its Bioactivities, Pharmacological Properties, and Potential as Alternative Medicine. Evid Based Complement Alternat Med. 2015;2015:919616.

Osthole, a coumarin compound, is a plant-based antifungal agent that primarily functions as a metabolite. It has been reported to be found in parsley (Seseli hartvigii), Japanese angelica (Angelica japonica), and other organisms with relevant data. See also: Angelica pubescens root (part).

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Therapeutic Uses
/EXPL THER/ Parsleyol is an active coumarin extracted from the dried fruit of Cnidium monnieri (L.) Cusson and is known to possess various pharmacological activities. This study aimed to investigate the mechanism by which parsleyol exerts a protective effect in an experimental model of allergic asthma. Our results showed that parsleyol treatment significantly reduced ovalbumin (OVA)-induced increases in serum IgE levels and inflammatory cytokines (IL-4, IL-5, IL-13) in bronchoalveolar lavage fluid (BALF), and decreased the recruitment of inflammatory cells in BALF and lung tissue. Furthermore, parsleyol effectively reduced goblet cell proliferation and excessive mucus secretion in lung tissue. Western blot analysis showed that parsleyol inhibited NF-κB activation, which may be related to reduced production of inflammatory cytokines. These data suggest that parsleyol alleviates OVA-induced allergic asthma inflammation by inhibiting NF-κB activation. This study elucidates the molecular mechanism of action of parsleyol, supporting its potential pharmacological application in the treatment of asthma and other airway inflammatory diseases. Hepatocellular carcinoma (HCC) accounts for approximately 90% of all primary liver cancer cases, and most HCC patients lack effective treatment options. Parsleyol is a traditional Chinese medicine reported to have various pharmacological functions, including hepatoprotective effects. In this study, we investigated the anticancer activity of parsleyol using HCC cell lines. We found that parsleyol inhibited HCC cell proliferation, induced cell cycle arrest, caused DNA damage, and inhibited HCC cell migration. Furthermore, we demonstrated that parsleyol not only promotes cell cycle G2/M phase arrest by downregulating the expression levels of Cdc2 and cyclin B1, but also induces DNA damage by increasing the expression of ERCC1. In addition, parsleyol inhibited the migration of hepatocellular carcinoma cell lines by significantly downregulating the levels of MMP-2 and MMP-9. Finally, we demonstrated that parsleyol inhibited epithelial-mesenchymal transition (EMT) by increasing the expression of epithelial biomarkers E-cadherin and β-catenin, and significantly decreasing the expression of mesenchymal biomarkers N-cadherin and vimentin. These results suggest that parsleyol may have potential chemotherapeutic activity for the treatment of hepatocellular carcinoma. Parsleyol (7-methoxy-8-isopentenoxycoumarin) is a compound extracted from Cnidium monnieri and has been found to have potent anticancer effects due to its ability to inhibit inflammation and cell proliferation. However, its effects on arterial wall hypertrophy-related diseases remain unclear. Therefore, this study aimed to investigate the effects of parsleyol on intimal hyperplasia in a rat carotid balloon injury model. We established a balloon-induced carotid artery injury model in male Sprague-Dawley rats, and then administered parsleyol (20 mg/kg/day or 40 mg/kg/day) or an equal volume of saline via gavage for 14 consecutive days. Subsequently, we assessed the degree of intimal hyperplasia and vascular smooth muscle cell proliferation by histopathological examination of carotid artery tissue and detection of proliferating cell nuclear antigen (PCNA) expression. Real-time quantitative RT-PCR was used to detect the expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), transforming growth factor-β1 (TGF-β1), and PCNA mRNA, and immunohistochemistry or Western blot was used to analyze the protein expression levels of nuclear factor-κB (NF-κB (p65)), IκB-α, TGF-β1, and phosphorylated Smad2 (p-Smad2). We found that parsleyol significantly alleviated neointimal thickening induced by balloon injury and reduced the increase in PCNA protein expression. Furthermore, parsleyol downregulated the upregulated expression of pro-inflammatory factors TNF-α, IL-1β, and NF-κB (p65) after balloon injury. Moreover, parsleyol treatment increased IκB-α protein expression. In addition, the balloon injury-induced increase in TGF-β1 and p-Smad2 protein expression was significantly inhibited by parsleyol. We conclude that parsleyol significantly inhibits neointimal hyperplasia in balloon-induced carotid artery injury in rats, and its mechanism may involve the downregulation of NF-κB, IL-1β, and TNF-α, thereby alleviating the inflammatory response, and the inhibition of the TGF-β1/Smad2 signaling pathway. Parsleyol has been shown to have various pharmacological effects, including antioxidant, anti-inflammatory, antiplatelet aggregation, estrogen-like effects, and analgesic effects. This study investigated the protective anti-inflammatory effect of parsleyol in a rat model of chronic renal failure (CRF) and its potential mechanism. Parsleyol treatment significantly reversed changes in serum creatinine, calcium, phosphorus, and blood urea nitrogen levels in CRF rats. Male Sprague-Dawley rats (8 weeks old) were given 200 mg/kg 2% adenine suspension (for the model group) to induce chronic renal failure (CRF). In the parsleyol treatment group, rats received 200 mg/kg 2% adenine suspension in combination with parsleyol (40 mg/kg, intravenously). Results showed that parsleyol treatment significantly inhibited the expression of tumor necrosis factor-α, interleukin (IL)-8, and IL-6 induced by CRF, and suppressed the expression of nuclear factor-κB (NF-κB) protein in CRF rats. Parsleyol treatment significantly reduced the protein expression of transforming growth factor-β1 (TGF-β1), decreased the activity of monocyte chemoattractant protein-1 (MCP-1), and increased the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) ratio in CRF rats. These results indicate that parsleyol exerts anti-inflammatory effects in a rat model of chronic renal failure (CRF) by inhibiting NF-κB and TGF-β1, and activating the PI3K/Akt/nuclear factor (erythrocyte-derived 2)-like 2 signaling pathway. Therefore, parsleyol may be a potential therapeutic agent for CRF. For more complete data on the therapeutic uses of parsleyol (out of 18), please visit the HSDB record page. Quercetin dihydrate is a natural flavonoid found in various fruits (apples, berries), vegetables (onions, broccoli), and herbs, possessing antioxidant, anti-inflammatory, and antitumor bioactivities [1,2,3].
- Its core mechanisms include: 1) scavenging reactive oxygen species (ROS) and enhancing the activity of antioxidant enzymes (superoxide dismutase/catalase) to alleviate oxidative stress; 2) inhibiting the NF-κB and PI3K/AKT signaling pathways, thereby inhibiting inflammation and tumor cell proliferation; 3) inducing tumor cell apoptosis and cell cycle arrest (G2/M phase) [1,2,3]
- It shows potential therapeutic value in inflammatory diseases (rheumatoid arthritis, acute inflammation), cancer (breast cancer, colon cancer), and age-related oxidative stress disorders (cognitive decline) [1,2,3]
- As a natural compound, it has good biocompatibility and low toxicity, making it a promising candidate for complementary and alternative medicine [1,3]

C15H16O3

244.29

244.109

C, 73.75; H, 6.60; O, 19.65

484-12-8

Osthole-d3

10228

White to off-white solid powder

1.1±0.1 g/cm3

396.7±42.0 °C at 760 mmHg

83-84°C

167.6±22.5 °C

0.0±0.9 mmHg at 25°C

1.557

3.87

0

3

3

18

366

0

O1C(C([H])=C([H])C2C([H])=C([H])C(=C(C1=2)C([H])([H])/C(/[H])=C(\C([H])([H])[H])/C([H])([H])[H])OC([H])([H])[H])=O

MBRLOUHOWLUMFF-UHFFFAOYSA-N

InChI=1S/C15H16O3/c1-10(2)4-7-12-13(17-3)8-5-11-6-9-14(16)18-15(11)12/h4-6,8-9H,7H2,1-3H3

7-methoxy-8-(3-methylbut-2-enyl)chromen-2-one

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)

Solubility in Formulation 1: ≥ 2.5 mg/mL (10.23 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 (10.23 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (10.23 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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 (Please use freshly prepared in vivo formulations for optimal results.)