OBJECTIVETo observe the oral acute toxicity of of (+)-usnic acid in mice and assess its cytotoxicity in rat cardiac fibroblasts.

METHODSThe mice with acute poisoning of (+)-usnic acid at different doses by oral administration were observed for toxic manifestations, and the LD(50) was determined. The survival time and survival rate of the mice receiving different doses of (+)-usnic acid were observed. Cultured rat cardiac fibroblasts were inoculated with different concentrations of (+)-usnic acid, and the cell growth inhibition rate was estimated and the IC(50) determined using MTT assay.

RESULTSHigher dose of (+)-usnic acid resulted in more obvious symptoms of poisoning and shorter survival time of the mice. The LD(50) of (+)-usnic acid in mice by oral administration was 388 mg/kg. The manifestations of poisoning such as apathism, pilomotor, chill, dyspnea, torpidity and anorexia was observed. Rat cardiac fibroblasts incubated with (+)-usnic acid showed obvious growth inhibition, which was positively correlated to the dose of (+)-usnic acid, and high dose of (+)-usnic acid caused severe cell injuries. The IC(50) of (+)-usnic acid in rat cardiac fibroblasts was 322 microg/ml.

CONCLUSION(+)-usnic acid is a natural compound of low toxicity in mice, and low to medium dose of (+)-usnic acid dose not produce obvious cytotoxicity.

Administration, Oral ; Animals ; Benzofurans ; administration & dosage ; chemistry ; toxicity ; Fibroblasts ; drug effects ; Lethal Dose 50 ; Mice ; Myocardium ; cytology ; Rats ; Stereoisomerism

BACKGROUNDDl-3-n-butylphthalide (NBP), first isolated from the seeds of celery, showed efficacy in animal models of stroke. This study was a clinical trial to assess the efficacy and safety of NBP with a continuous dose regimen among patients with acute ischemic stroke.

METHODSA randomized, double-blind, double-dummy trial enrolled 573 patients within 48 hours of onset of ischemic stroke in China. Patients were randomly assigned to receive a 14-day infusion of NBP followed by an NBP capsule, a 14-day infusion of NBP followed by aspirin, or a 14-day infusion of ozagrel followed by aspirin. The efficacy measures were Barthel index score and the modified Rankin scale (mRS) at day 90. Differences among the three groups on mRS were compared using χ(2) test of proportions (with two-sided α = 0.05) and Logistic regression analysis was conducted to take the baseline National Institutes of Health Stroke Scale (NIHSS) score into consideration.

RESULTSAmong the 535 subjects included in the efficacy analysis, 90-day treatment with NBP was associated with a significantly favorable outcome than 14-day treatment with ozagrel as measured by mRS (P < 0.001). No significant difference was found among the three groups on Barthel index at day 90. The rate of adverse events was similar among the three groups.

CONCLUSIONSThe 90-day treatment with NBP could improve outcomes at the third month after stroke. The NBP treatment (both intravenous and oral) is safe (ChiCTR-TRC-09000483).

Adult ; Aged ; Benzofurans ; administration & dosage ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Male ; Middle Aged ; Stroke ; drug therapy ; Treatment Outcome

OBJECTIVETo study the relationship between deposition content and time of the active ingredients in rat skin, and investigate the dermatopharmacokinetics of Liangfu Cream.

METHODThe contents of paeonol, dictamnine, fraxinellone and glycyrrhetinic acid in rat skin were determined by HPLC. The dermatopharmacokinetics parameters were calculated by DAS software.

RESULTThe dermatopharmacokinetics of paeonol and glycyrrhetinic acid were two compartment model, while that of dictamnine and fraxinellone were one compartment model: T(1/2Ka) of four active ingredients were 0.307, 0.112, 0.146, 0.216 h, respectively; T(lag) of them were 0.006, 0.123, 0.136, 0.109 h, respectively; all the Tmax of them was 0.5 h; the Cmax, were 40.163, 1.607, 6.725, 100.553 microg x cm(-3), respectively; the t(1/2beta), were 14.719, 1.262, 0.838, 234.807 h, respectively; the AUC(0-infinity), were 16.987, 2.713, 9.345, 697.000 microg x cm(-3) x h(-1), respectively; and the MRT(0-infinity) were 3.662, 1.67, 1.585, 10.897, respectively.

CONCLUSIONThe skin pharmacokinetics characteristic of four ingredients in Liangfu cream is lined with the cataplasm long time.

Acetophenones ; administration & dosage ; pharmacokinetics ; Administration, Cutaneous ; Animals ; Benzofurans ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glycyrrhetinic Acid ; administration & dosage ; pharmacokinetics ; Male ; Mice ; Quinolines ; administration & dosage ; pharmacokinetics ; Skin ; drug effects

AIM: To identify and quantify the major metabolites of salvianolic acid B (SAB) after intravenous injection in rats. METHODS: LC-IT/TOF-MS was used to identify the metabolites in rat bile, plasma, and urine; LC-MS/MS was used to quantify the two major metabolites. RESULTS: In rat bile, plasma, and urine, nine metabolites were identified, including methylated metabolites of SAB, lithospermic acid (LSA), the decarboxylation and methylation metabolites of LSA, salvianolic acid S (SAS), and dehydrated-SAS. The t1/2 of monomethyl-SAB and LSA were both very short, and monomethyl-SAB had a larger AUC than LSA in rats. CONCLUSION Nine metabolites were found, the metabolic pathway was described, and the pharmacokinetic profiles of LSA and monomethyl-SAB were studied, thereby clarifying that methylation was the dominant metabolic pathway for SAB in rats.
Administration, Intravenous ; Animals ; Benzofurans ; administration & dosage ; chemistry ; metabolism ; pharmacokinetics ; Bile ; chemistry ; Male ; Molecular Structure ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Urine ; chemistry

BACKGROUNDAmyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by progressive death of the upper and lower motor neurons. Transgenic mice over-expressing a mutant form of the human SOD1 gene develop an ALS-like phenotype. Currently, there is no effective treatment or drug for the fatal disease. Previous studies reported potent efficacy of dl-3-n-butylphthalide (DL-NBP) for several neurodegenerative disorders and cerebral ischemia. SOD1-G93A mice are a mouse model of ALS. In this study, we investigated the efficacy of DL-NBP on this ALS mouse model.

METHODSSixty SOD1-G93A female mice were divided into four groups. The vehicle control group received 0 mg×kg(-1)×d(-1) DL-NBP. The experimental groups received DL-NBP with doses of 30, 60 or 120 mg×kg(-1)×d(-1), respectively. For measurement of motor activity, the hanging wire test and rotarod test were performed. Survival statistics were analyzed by Kaplan-Meier survival curves. The body weight of each mouse was recorded twice per week. The statistical motor unit number estimation (MUNE) technique was used to estimate the number of functioning motor units in gastrocnemius muscle. Muscle morphology was evaluated by hematoxylin and eosin staining. Motor neuron quantitation was performed by Nissl staining and microglia activation was observed by immunohistochemistry.

RESULTSOral administration of 60 mg×kg(-1)×d(-1)1 DL-NBP significantly prolonged survival ((164.78 ± 16.67) days) of SOD1-G93A mice compared with vehicle control ((140.00 ± 16.89) days). Treating mice with DL-NBP (60 mg×kg(-1)×d(-1)) significantly decreased the progression rate of motor deficits and suppressed body weight reduction. Furthermore, we found that treating SOD1-G93A mice with DL-NBP (60 mg×kg(-1)×d(-1)) slowed the rate of MUNE reduction (P < 0.01). Motor neurons were remarkably preserved in the anterior horns in mice treated with DL-NBP (60 mg×kg(-1)×d(-1)) at the stage of 19 weeks (P < 0.01). Treating mice with DL-NBP (60 mg×kg(-1)×d(-1)) significantly reduced CD11b immunoreactivity compared with vehicle control mice (P < 0.05). No significant effect was observed in mice treated with DL-NBP of 30 or 120 mg×kg(-1)×d(-1).

CONCLUSIONSThe post-disease-onset administration of DL-NBP significantly prolonged survival and improved motor performance in SOD1-G93A mice. DL-NBP may be a potential therapeutic agent for ALS.

Administration, Oral ; Amyotrophic Lateral Sclerosis ; drug therapy ; Animals ; Benzofurans ; administration & dosage ; therapeutic use ; Disease Models, Animal ; Female ; Immunohistochemistry ; Mice ; Mice, Transgenic

Coumarins are the main active components in Psoraleae Fructus. To study the multi-component pharmacokinetics of Psoraleae Fructus, this study established a sensitive and rapid ultra-pressure liquid chromatography coupled to tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of psoralen, isopsoralen, psoralenoside, and isopsoralenoside in rat plasma. After validation, the method was applied to the investigation of pharmacokinetics of psoralen, isopsoralen, psoralenoside, and isopso-ralenoside in rats after single and multiple administration of Psoraleae Fructus extract. The results revealed that the exposure of psoralen and isopsoralen in rat plasma was high after a single intragastric administration of Psoraleae Fructus extract, with an AUC_(0-∞) of 443 619-582 680 and 167 314-276 903 ng·mL~(-1)·h~(-1), respectively. Compared with these two compounds, the exposure of psoralenoside and isopsoralenoside was lower with marked gender difference. After 7-day administration of Psoraleae Fructus extract to rats, the AUC_(0-∞) of psoralen and isopsoralen was 29 701-81 783 and 39 234-89 914 ng·mL~(-1)·h~(-1), respectively, which was significantly lower than that at the first day(P<0.05), and that of psoralenoside and isopsoralenoside was 7 360-19 342 and 8 823-45 501 ng·mL~(-1)·h~(-1), respectively. There was no significant gender difference in exposure of psoralenoside and isopsoralenoside in male and female rats. However, the exposure of psoralenoside and isopsoralenoside in male rats was reduced(P<0.05), and the t_(1/2) and mean residence time(MRT) were shortened, suggesting that the removal of these two compounds from the body was accelerated.
Administration, Oral ; Animals ; Benzofurans ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drugs, Chinese Herbal ; Ficusin ; Furocoumarins/analysis* ; Glycosides ; Psoralea ; Rats ; Tandem Mass Spectrometry

OBJECTIVETo observe the effect of psoralidine in rats with ovariectomy, and preliminarily study its mechanism.

METHODSixty female Sprague-Dawley rats were divided into 5 groups: the sham operation group, the model group, the psoralidine low-dose group (4 mg x kg(-1)), the psoralidine high-dose group (16 mg x kg(-1)) and the Zhuangguzhitong capsule group, with 12 rats in each group. Thirteen weeks later, their blood and bone samples were collected to detect bone density, bone biochemistry, pathomorphology, serum E2 and CT.

RESULTPsoralidine could up-regulate the bone density of lumbar vertebra and thighbone of rats with ovariectomy (P < 0.05), the maximum bending strength of thighbone (P < 0.05), and serum E2 and CT (P < 0.05).

CONCLUSIONPsoralidine has a good active effect on postmenopausal antiosteoporosis. Its mechanism may be related to such pathways as E2 and CT.

Animals ; Benzofurans ; administration & dosage ; Bone Density ; drug effects ; Coumarins ; administration & dosage ; Drugs, Chinese Herbal ; administration & dosage ; Estradiol ; blood ; Female ; Humans ; Osteoporosis, Postmenopausal ; blood ; drug therapy ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the protective effect and mechanism of salvianolic acid B on isolated heart ischemia/reperfusion injury in rats.

METHODForty-eight SD rats were divided into 6 groups randomly(n = 8): the control group, the positive administration group (verapamil 150 microg x L(-1)), and high, middle and low-dose salvianolic acid B groups (10, 5, 2.5 mg x L(-1)). The myocardial ischemia/reperfusion injury model was established using the Langendorff method, re-perusing isolated working hearts for 30 min after ischemia for 25 min. A water-bag catheter was inserted in rat left atrium for recording the effect of salvianolic acid B on hemodynamics indexes-AST, LDH, SOD and MDA.

RESULTVarious group with different doses showed that salvianolic acid B decreased AST, release of LDH and formation of MDA and increased SOD activity.

CONCLUSIONSalvianolic acid B showed a protective effect on myocardial ischemia/reperfusion injury. Its mechanism may be related with improvement of cardiac contractility, cleaning of oxygen free radicals and reduction of lipid peroxidation.

Animals ; Benzofurans ; administration & dosage ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Hemodynamics ; drug effects ; In Vitro Techniques ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Time Factors

OBJECTIVETo study the effects of different penetration enhancers on the in vitro percutaneous absorption and amount retained in skin of active ingredients in Liangfu cream and to screen out the effective accelerator.

METHODUsing improved Franz-type difusion cell and excised small mouse skin in vitro as transdermal barrier, the amount retained in skin and kinetics parameters of active ingredients such as cumulative permeation quantity, permeation rate and permeation lagged time were determined by HPLC. The enhancement ability of four different enhancers such as azone, oleic acid, transcutol P and isopropyl myristate were investigated.

RESULT3% IPM enhanced the cumulative permeation quantity better than other penetration enhancers. The enhancive permeation multiples of paeonol, dictamnine, fraxinellone and glycyrrhetinic acid were 1.52, 1.24, 1.73 and 3.21 times (P < 0.05). The enhancive amount retained in skin multiple of glycyrrhetinic acid was 1.96 times (P < 0.05), but for other components there were no significant impacts.

CONCLUSIONThe effects of penetration enhancers on the in vitro percutaneous absorption and amount retained in skin of components in Liangfu cream are different. 3% IPM which can enhance the cumulative permeation quantity of four components and amount retained in skin of glycyrrhetinic acid is the most suitable penetration enhancer for Liangfu cream.

Acetophenones ; chemistry ; Administration, Cutaneous ; Animals ; Benzofurans ; chemistry ; Drug Carriers ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glycyrrhetinic Acid ; chemistry ; Male ; Mice ; Quinolines ; chemistry ; Skin ; drug effects ; Skin Absorption ; drug effects

OBJECTIVETo develop a LC-MS method for the determination of senkyunolide I (SI) in rat plasma, in order to observe whether there is significant change in the pharmacokinetics parameters of complex prescriptions of Huoluoxiaolingdan (HLXL) and single herbal extracts from Ligusticum chuanxiong Hort. in rats, and assess the effect of other components in HLXL on the pharmacokinetics of SI.

METHODTwelve male Sprague-Dawley (SD) rats were randomly divided into two groups, and orally administered with extract from HLXL and L. chuanxiong (both equal to SI 4.53 mg x kg(-1)). Their blood was collected at different time points for LC-MS, in order to detect the plasma concentration of SI. The pharmacokinetic parameters of SI were calculated by DAS 2.0 software. SPSS 16.0 software was used for independent-sample T-test and Nonparametric T-test.

RESULTA linear relationship of SI ranged from 6.750 to 675.0 microg x L(-1), and with the lowest limit of detection being 6.750 microg L(-1). Both of the plasma concentration-time curves of SI were fitted with the two-compartment model for extract of HLXL and L. chuanxiong. The detected AUC and Cmax of SI showed significant difference, with no significant difference in other parameters.

CONCLUSIONThe LC-MS determination method established in this experiment was so exclusive, accurate and sensitive that it is suitable for pharmacokinetic studies on extracts of HLXL and SI from L. chuanxion. The experiment results show that other ingredients of HLXL have noticeable effect on the absorption of SI in rat plasma.

Administration, Oral ; Animals ; Area Under Curve ; Benzofurans ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Metabolic Clearance Rate ; Random Allocation ; Rats ; Rats, Sprague-Dawley