Psoriasis is an auto-immune skin disease, which is characterized by the excessive generation of plaques on the skin with typically a long-lasting red, itchy and scaly symptoms. Imiquimod, which has been used for the treatment of external genital warts, actinic keratosis, and superficial basal cell carcinoma, induced of psoriasis-like skin disorders with skin erythema and thickness in mice. In the present study, we tried to find the bioactive herbal extract against imiquimod-induced psoriasis-like skin disorder in mice. During the searching of the herbal extract with anti-psoriatic effect, the ethanolic extract of Cnidium officinale ameliorated imiquimodinduced psoriasis-like skin disorder in mice. The morphological evaluation, H&E staining and Psoriasis Area and Severity Index (PASI) score showed that ear and back thickness, and erythema induced by imiquimod were significantly reversed after the treatment of the cream of the ethanolic extract of C. officinale. The overexpressed myeloperoxidase (MPO) and keratin 6A levels were decreased by the treatment of C. officinale cream. Also, IFN-γ, c-fos and IκB-α mRNA levels, which are related to the progression of psoriasis, were reduced by C. officinale cream. Thus, the ethanolic extract of C. officinale ameliorated psoriasis-like skin disorder induced by imiquimod and might be the therapeutic agent for psoriasis.
Animals ; Carcinoma, Basal Cell ; Cnidium ; Condylomata Acuminata ; Ear ; Erythema ; Ethanol ; Keratin-6 ; Keratosis, Actinic ; Mice ; Peroxidase ; Psoriasis ; RNA, Messenger ; Skin Diseases ; Skin

There is an increasing interest in phytoestrogens due to their potential medical usage in hormone replacement therapy (HRT). The present study was designed to investigate the in vitro effects of estrogen-like activities of two widespread coumarins, osthole and imperatorin, using the MCF-7 cell proliferation assay and their alkaline phosphatase (ALP) activities in osteoblasts Saos-2 cells. The two compounds were found to strongly stimulate the proliferation of MCF-7 cells. The estrogen receptor-regulated ERα, progesterone receptor (PR) and PS2 mRNA levels were increased by treatment with osthole and imperatorin. All these effects were significantly inhibited by the specific estrogen receptor antagonist ICI182, 780. Cell cycle analysis revealed that their proliferation stimulatory effect was associated with a marked increase in the number of MCF-7 cells in S phase, which was similar to that observed with estradiol. It was also observed that they significantly increased ALP activity, which was reversed by ICI182,780. These results suggested that osthole and imperatorin could stimulate osteoblastic activity by displaying estrogenic properties or through the ER pathway. In conclusion, osthole and imperatorin may represent new pharmacological tools for the treatment of osteoporosis.
Alkaline Phosphatase ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cnidium ; chemistry ; Coumarins ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Furocoumarins ; pharmacology ; Humans ; MCF-7 Cells ; Osteoblasts ; cytology ; drug effects ; enzymology ; Phytoestrogens ; pharmacology ; Receptors, Estrogen ; genetics ; metabolism

Although the effects of the rice bran have recently been investigated, there is no information regarding platelet physiology available. However, it is well known that fermented natural plants have a beneficial effect on cardiovascular diseases. Therefore, this study was conducted to investigate whether fermented rice bran extract (FRBE) with several plants (Artemisia princeps, Angelica Gigantis Radix, Cnidium officinale, and Camellia sinensis) affected agonist-induced platelet aggregation, and if so, what the underlying mechanism of its activity was. We performed several experiments, including in vitro platelet aggregation, intracellular calcium concentration and adenosine triphosphate release. In addition, the activation of integrin alphaIIbbeta3 was determined using fibrinogen binding. Thrombus formation was also evaluated in vivo using an arterio-venous shunt model. The FRBE inhibited collagen-induced platelet aggregation in a concentration-dependent manner. FRBE significantly and dose dependently attenuated thrombus formation using rat arterio-venous shunt. FRBE suppressed the intracellular calcium mobilization in collagen-stimulated platelets. We also found that FRBE inhibited extracellular stimuli-responsive kinase 1/2, p38-mitogen-activated protein kinases and c-Jun N-terminal kinase phosphorylation. These results suggested that FRBE inhibited collagen-induced platelet aggregation, which was mediated by modulation of downstream signaling molecules. In conclusion, FRBE could be developed as a functional food against aberrant platelet activation-related cardiovascular diseases.
Adenosine Triphosphate ; Angelica ; Animals ; Blood Platelets ; Calcium ; Camellia ; Cardiovascular Diseases ; Cnidium ; Collagen ; Fibrinogen ; Functional Food ; JNK Mitogen-Activated Protein Kinases ; Phosphorylation ; Phosphotransferases ; Physiology ; Platelet Aggregation ; Platelet Glycoprotein GPIIb-IIIa Complex ; Protein Kinases ; Rats ; Thrombosis

This study is to study is to investigate the coumarins from Fruit of Cnidium monnieri and their cytotoxic activities. The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for their cytoxic activities by MTT method. Eleven compounds were isolated and identified as osthole (1), bergaptan (2), xanthotoxol (3), xanthotoxin (4), imperatorin (5), isopimpinellin (6), osthenol (7), psoralen (8), 5,7-dimethoxycoumarin (9), oxypeucedaninhydrate (10), and swietenocoumarin F (11). Compounds 7, 9-11 were isolated from the Cnidium genus for the first time. Compounds 1,5,10 and 11 showed significant cytotoxic activities against L1210 cell lines at a concentration of 1 x 10(-5) mol x L(-1) with inhibitory rates of were 70.13, 63.10, 55.77, and 75.08% respectively.
Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cnidium ; chemistry ; toxicity ; Coumarins ; chemistry ; isolation & purification ; toxicity ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; toxicity ; Fruit ; chemistry ; toxicity ; Mice ; Molecular Structure

The aim of this study was to investigate the inhibitory effect of Cnidium monnieri fruit (CM) extracts on pulmonary inflammation induced in mice by cigarette smoke condensate (CSC) and lipopolysaccharide (LPS). Pulmonary inflammation was induced by intratracheal instillation of LPS and CSC five times within 12 days. CM extract was administered orally at a dose of 50 or 200 mg·kg(-1). The number of inflammatory cells in the bronchoalveolar lavage fluid was counted using a fluorescence activated cell sorter. Inflammatory mediator levels were determined by enzyme-linked immunosorbent assay. The administration of LPS and CSC exacerbated airway hyper-responsiveness (AHR) and induced an accumulation of inflammatory cells and mediators, and led to histological changes. However, these responses are modulated by treatment with CM, and the treatment with CM extract produces similar or more extensive results than the treatment with cyclosporin A (CSA). CM extract may have an inhibitory effect on pulmonary inflammation related with chronic obstructive pulmonary disease.
Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Bronchoalveolar Lavage Fluid ; Cnidium ; Female ; Fruit ; Lipopolysaccharides ; Mice, Inbred BALB C ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Pneumonia ; chemically induced ; drug therapy ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; pathology ; Smoke ; adverse effects ; Smoking ; adverse effects ; Tobacco Products ; adverse effects

This study is to investigate the effect of osthol on osteoclasts' activity, bone resorption as well as apoptosis in vitro, and explore the mechanism of osthol in preventing osteoporosis. Osteoclasts were separated from long-limb bones of new born rabbits, cultured in 24-well plate with glass slices and bone slices, and treated by 1 x 10(-5) mol x L(-1) osthol. Osteoclasts were identified by observing live cells with phase contrast microscope, HE staining, TRAP staining and toluidine blue staining of bone resorption pits. The numbers of bone resorption pits were counted as well as the surface area of bone resorption on bone slice. Osteoclasts were stained with acridine orange to detect the cell apoptosis. The ratio of apoptotic osteoclasts was observed under fluorescence microscope. The gene expression of RANKL, OPG, TRAP and p-JNK1/2 protein expression were examined using real time PCR and Western blotting, respectively. Comparing with the control group without osthol, the rates of apoptotic osteoclasts increased obviously and the number and area of bone resorption pits decreased evidently with 1 x 10(-5) mol x L(-1) osthol. There is significant difference between control group and experiment group treated by 1 x 10(-5) mol x L(-1) osthol. Therefore, the osthol through RANK+RANKL/TRAF6/Mkk/JNK signal pathway inhibits the osteoclasts activity, enhances osteoclasts apoptotic and inhibits the bone resorption.
Acid Phosphatase ; metabolism ; Animals ; Apoptosis ; drug effects ; Bone Resorption ; Cells, Cultured ; Cnidium ; chemistry ; Coumarins ; isolation & purification ; pharmacology ; Gene Expression ; Isoenzymes ; metabolism ; Mitogen-Activated Protein Kinase 8 ; metabolism ; Mitogen-Activated Protein Kinase 9 ; metabolism ; Osteoclasts ; metabolism ; pathology ; Osteoprotegerin ; metabolism ; Phosphorylation ; Plants, Medicinal ; chemistry ; RANK Ligand ; metabolism ; Rabbits ; Seeds ; chemistry ; Signal Transduction ; Tartrate-Resistant Acid Phosphatase

This paper is to study the inhibitory effect of water soluble polymers--methyl cellulose (MC), hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC-M), poloxamer (F68) and polyvidon (PVP) on osthole (OST) crystallization and investigate the impact of polymer concentration and viscosity on crystallization behavior. Also, UV spectrophotometry method was used to determine the drug concentration at different time point to draw the OST concentration-time curve. Results show that HPMC has the most significant inhibition effect on OST crystallization, and drug concentration level is 1.61 times higher than that in control solution within 8 h followed by PVP (1.54) and MC (1.45) respectively. The kinetics of OST recrystallization can be described using first-order reaction, and the crystallization rate constants obtained by analyzing the regression equation indicate that HPMC-60SH-4000 and HPMC-60SH-10000 can greatly influence OST crystal formation. The dissolution rate of drugs precipitated from water-soluble polymer solutions is faster compared with controls in pH 1.2 HCl and pH 6.8 phosphate buffers, which demonstrated that water-soluble polymers can not only change the behavior of drug crystallization but markedly improve the dissolution rate of water insoluble drugs.
Cellulose ; analogs & derivatives ; chemistry ; Cnidium ; chemistry ; Coumarins ; chemistry ; isolation & purification ; Crystallization ; Hypromellose Derivatives ; Kinetics ; Methylcellulose ; analogs & derivatives ; chemistry ; Plants, Medicinal ; chemistry ; Poloxamer ; chemistry ; Polymers ; chemistry ; Povidone ; chemistry ; Solubility ; Viscosity

The paper is aimed to study the metabolic characteristics of osthol (Ost) in isolated hepatocytes of rat to identify which isoforms of CYP450 were responsible for Ost metabolism in vitro. The concentration of Ost in isolated hepatocytes incubation system was determined by HPLC-UV. The effects of incubation time, substrate concentration and hepatocytes amount on the metabolic characteristics of Ost were investigated. CYP2C8 inhibitor quercetin (Que), CYP2C9 inhibitor sulfaphenazole (Sul), CYP2D6 inhibitor yohimbine (Yoh), CYP3A4 inhibitor troleandomycin (Tro) and CYP450 inducer rifampicin (Rif) were used to investigate their effects on the metabolism of Ost. The metabolism of Ost in isolated rat hepatocytes showed an enzymatic kinetic characteristics. Rif induced Ost elimination in rat hepatocytes; Yoh, Sul, Que did not have effects on Ost metabolism in vitro. Between 0-200 micromol x L(-1), Tro inhibited Ost metabolism in a concentration-dependent manner. CYP3A4 is the enzyme metabolizing Ost in vitro; CYP2C8, CYP2C9 and CYP2D6 did not involve in Ost metabolism in rat hepatocytes.
Animals ; Cells, Cultured ; Cnidium ; chemistry ; Coumarins ; isolation & purification ; metabolism ; Cytochrome P-450 CYP2D6 Inhibitors ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; Hepatocytes ; metabolism ; Male ; Plants, Medicinal ; chemistry ; Quercetin ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rifampin ; pharmacology ; Sulfaphenazole ; pharmacology ; Troleandomycin ; administration & dosage ; pharmacology ; Yohimbine ; pharmacology

To establish the method of HPLC-fingerprint analysis for the quality control of Cnidium monnieri L. Cuss., and identify its active constituents by HPLC-MS, 35 batches of samples were analyzed on a Shimadzu C18 column with a gradient of acetonitrile and 0.1% aqueous aceticacid at a flow rate of 1.0 mL x min(-1) and detected at 245 nm and 322 nm. Furthermore, the typical samples were detected by HPLC-DAD-MS under negative ion mode. 35 batches of Cnidium monnieri L. Cuss. samples were classified into four types based on the results of similarity analysis. According to the comparison of the t(R), MS data and UV maximum absorbance (lambda(max)) values with the standards, 8, 7, 4 and 2 coumarins components were identified in four types of Cnidium monnieri L. Cuss. extracts, separately. The method is repeatable and reliable, and it is capable of effectively controlling the quality of Cnidium monnieri L.
Chromatography, High Pressure Liquid ; methods ; Cnidium ; chemistry ; Coumarins ; analysis ; Ecosystem ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Seeds ; chemistry ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; Spectrophotometry, Ultraviolet

OBJECTIVETo develop an RP-HPLC method for simultaneous determination of 5 constituents in Fructus Cnidii.

METHODAnalysis was performed on an Alltech C18 (4.6 mm x 250 mm, 5 microm) column. The mobile phases were acetonitrile water and acetic acid with gradient elution. The flow rate was 1 mL x min(-1). The monitoring wavelength was 325 nm and 245 nm. The column temperature was 40 degrees C.

RESULTThe linear response ranges were 1-20 microg x mL(-1) (r = 0.999 9) for xanthotoxin, 1-20 microg x mL(-1) (r = 0.999 9) for isopimpinellin, 11-20 microg x mL(-1) (r = 0.999 8) for bergapten, 100-1 200 microg x mL(-1) (r = 0.999 7) for imperatorin, 100-2 000 microg x mL(-1) (r = 0.999 9) for osthole. The average recoveries were all above 95%.

CONCLUSIONThe method is simple, sensitive and accurate with good reproducibility.

Chromatography, High Pressure Liquid ; methods ; Cnidium ; chemistry ; Coumarins ; analysis ; Fruit ; chemistry ; Furocoumarins ; analysis ; Methoxsalen ; analogs & derivatives ; analysis ; Plants, Medicinal ; chemistry ; Reproducibility of Results