OBJECTIVETo prepare polylactic acid microspheres containing total alkaloid extracts of Caulis sinomenii and study their release characteristics in vitro.

METHODPolylactic acid microspheres containing total alkaloid extracts of C. sinomenii were prepared by O/W emulsification solvent-evaporation process. The microspheres were characterized in terms of morphology, encapsulation efficiency, and particle size distribution. The effect of different conditions on release property of microspheres was studied.

RESULTThe formed microspheres were spherical with smooth surfaces. The encapsulation efficiency and rate of drug loading were (83.4 +/- 5.63)% and (8.7-0.35)%, respectively. The distribution of particle size was uniform and the average size was (21.5 +/- 1.22) microm. In vitro release study revealed that the 32-hour accumulative release percentage reached 80%.

CONCLUSIONPolylactic acid microspheres containing total alkaloid extracts of C. sinomenii were prepared successfully. Microspheres with good sustained-release characteristics can be produced by controlling different process parameters.

Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Lactic Acid ; chemistry ; Microspheres ; Polyesters ; Polymers ; chemistry ; Sinomenium ; chemistry

OBJECTIVETo explore the material basis of Caulis sinomenii (CS) in treating osteoarthritis (OA), and to give a pharmacodynamic illustration for the multi-targeting therapeutics of CS.

METHODSThe computational methods, consisting of molecular docking and biological network were carried out to search the database targeting twelve important OA related enzymes: ASAMTS4, ASAMTS5, MMP-1, MMP-3, MMP-13, MMP-8, MMP-2, COX-2, COX-1, IL-1beta, TNF-alpha, iNOS, and map the ligand-target interaction networks about molecules from CS and DrugBank. After that, an aggregate analysis was performed to analyze the mechanisms of compositions in CS.

RESULTSTotally 14 had good interaction in all molecules in database with two or more than two of the OA correlated enzymes, and 6 molecules had interaction with four or more enzymes. Moreover, both herb ligand-target interaction network and drug ligand-target interaction network were similar in the interaction profiles and network features, which revealed multi-drugs effects in CS.

CONCLUSIONSThere were a lot of multi-target molecules in CS, providing pharmacodynamic illustrations for the multi-target therapeutics of Chinese medicine. Meanwhile, they supplied certain reference and inspiration for finding out new drugs for OA therapy.

Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Molecular Docking Simulation ; Osteoarthritis ; drug therapy ; Phytotherapy ; Sinomenium

OBJECTIVE: To decipher the possible mechanisms of Sinomenium Acutum (SA) in treating diseases by a bioinformatics method. METHODS: SA ingredients were searched according to Chinese Pharmacopoeia, Chinese Medicine Dictionary and Traditional Chinese Medicines Database (TCMD). Active compounds and target proteins of SA were acquired through the Pubchem platform. Pathway, network and function analyses of SA were performed with ingenuity pathway analysis (IPA), a bioinformatics analysis platform. Disease, biofunction-target networks were established with Cytoscape. RESULTS: Eighteen ingredients from SA were obtained. Seven active ingredients with 31 active target proteins were acquired according to PubChem Bioassay test. By IPA analysis, 277 canonical pathways belonging to 17 function categories were collected, 23 kinds of diseases, 21 categories bio-functions were obtained. Based on P value, calculated by IPA, the top 5 significant pathway of SA targets include phosphatidylinositol 3 kinase/Akt (PI3K/Akt) signaling, prostate cancer signaling, macrophage migration inhibitory factor (MIF) regulation of innate immunity, Guanosine-binding protein coupled receptor (GPCR) signaling, and ataxia telangiectasia mutated protein (ATM) signaling. Disease and bio-function network analysis indicated that mitogen activated protein kinase 1 (MAPK1), MAPK3, p65 nuclear factor κB (RELA), nuclear factor of κB inhibitor alpha (NFκBIA), interleukin 1β(IL-1β), prostaglandin G/H synthase 2 (PTGS2) and tumor protein 53 (TP53) were the critical targets in various diseases treated by SA. CONCLUSION In the different view of target, pathway, disease and bio-function, inflammation was found to be a central theme in many chronic conditions. SA could be used not only as an anti-inflammatory agent, but also for the treatment of cancers, neurological diseases, psychological disorders and metabolic diseases.
Computational Biology ; methods ; Disease ; Humans ; Molecular Targeted Therapy ; Proteins ; metabolism ; Signal Transduction ; Sinomenium ; chemistry

OBJECTIVETo establish an HPLC method for the determination of entrapment efficiency of sinomenine liposomes.

METHODThe liposomes and dissociated drugs were separated by sephadex filtration, mini-column centrifugation and dialysis. The methodology study and the optimization of determining condition were carried out at the same time.

RESULTSephadex filtration could effectively separate the sinomenine liposomes from dissociated sinomenine. The column recovery was 98.8%, the average entrapment efficiency of three tests was64.9%, RSD 2.67%.

CONCLUSIONThe method was simple, exact, and had a good reappearance. It can be used to examine the entrapment efficiency of sinomenine liposomes.

Dextrans ; Drug Carriers ; Drug Delivery Systems ; Filtration ; methods ; Liposomes ; Morphinans ; administration & dosage ; analysis ; isolation & purification ; Sinomenium ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo prepare Sinomenine hydrochloride-loaded bovine serum albumin microspheres (SM-BSA-MS).

METHODSM-BSA-MS was prepared by spray drying technique. The morphology, drug-loading and release in vitro of SM-BSA-MS was studied.

RESULTThe diameters of SM-BSA-MS were in the range of 1-3 m. The drug loading of microspheres, formulated with different drug/albumin ratios as 1, 2, 1:1, 2:1, were 31.6%, 47.7% and 67.9% , respectively. And the drug entrapment efficiencies of different drug/albumin ratios were higher than 94%. The results of in vitro release experiments showed that the drug loaded microspheres have the properties of sustained-release compared with the Sinomenine hydrochloride injection. Different release characteristics could be obtained by adjusting the prescription composition and the thermal denaturation condition.

CONCLUSIONSpray drying technique is a simple and feasible method for preparing SM-BSA-MS. The drug loaded microspheres had high drug-loading and sustained-release effect.

Delayed-Action Preparations ; chemistry ; Drug Carriers ; Drug Compounding ; methods ; Microspheres ; Morphinans ; chemistry ; Particle Size ; Plants, Medicinal ; chemistry ; Serum Albumin, Bovine ; chemistry ; Sinomenium ; chemistry

AIMTo determine in vitro the rat plasma protein binding rate by using microdialysis method.

METHODSThe binding rate was determined by using microdialysis probe as sampling tools and zero-net flux method as calibrating method. The regression equation was made by the difference of concentrations between the dialysis sample and the perfusate. The x-intercept of regression equation was the free drug concentration (Cf). The plasma protein binding rate was calculated by using the following equation: f = ( C0 - Cf)/C0.

RESULTThe binding rate was kept relatively stable in the studied concentration range.

CONCLUSIONIt is feasible that the plasma protein binding rate can be determined by using microdialysis method.

Animals ; Blood Proteins ; metabolism ; Chromatography, High Pressure Liquid ; Male ; Microdialysis ; methods ; Morphinans ; isolation & purification ; metabolism ; Plants, Medicinal ; chemistry ; Protein Binding ; Rats ; Rats, Sprague-Dawley ; Regression Analysis ; Sinomenium ; chemistry

Sinomenium acutum has been long used in the preparations of traditional medicine in Japan, China and Korea for the treatment of various disorders including rheumatism, fever, pulmonary diseases and mood disorders. Recently, it was reported that Sinomenium acutum, has sedative and anxiolytic effects mediated by GABA-ergic systems. These experiments were performed to investigate whether sinomenine (SIN), an alkaloid derived from Sinomenium acutum enhances pentobarbital-induced sleep via γ-aminobutyric acid (GABA)-ergic systems, and modulates sleep architecture in mice. Oral administration of SIN (40 mg/kg) markedly reduced spontaneous locomotor activity, similar to diazepam (a benzodiazepine agonist) in mice. SIN shortened sleep latency, and increased total sleep time in a dose-dependent manner when co-administrated with pentobarbital (42 mg/kg, i.p.). SIN also increased the number of sleeping mice and total sleep time by concomitant administration with the sub-hypnotic dosage of pentobarbital (28 mg/kg, i.p.). SIN reduced the number of sleep-wake cycles, and increased total sleep time and non-rapid eye movement (NREM) sleep. In addition, SIN also increased chloride influx in the primary cultured hypothalamic neuronal cells. Furthermore, protein overexpression of glutamic acid decarboxylase (GAD(65/67)) and GABA(A) receptor subunits by western blot were found, being activated by SIN. In conclusion, SIN augments pentobarbital-induced sleeping behaviors through GABA(A)-ergic systems, and increased NREM sleep. It could be a candidate for the treatment of insomnia.
Administration, Oral ; Animals ; Anti-Anxiety Agents ; Benzodiazepines ; Blotting, Western ; China ; Diazepam ; Eye Movements* ; Fever ; Glutamate Decarboxylase ; Japan ; Korea ; Lung Diseases ; Medicine, Traditional ; Mice ; Mood Disorders ; Motor Activity ; Neurons ; Pentobarbital ; Receptors, GABA-A ; Rheumatic Diseases ; Rodentia* ; Sinomenium* ; Sleep Initiation and Maintenance Disorders

OBJECTIVETo study the effects of sinomenine (Sin) on cell proliferation, intracellular expression of cyclooxygenase-2 (COX-2), and production of PGE2 in lipopolysaccharide-induced PC-12 cells, To explore the Sin's mechanism on nerve cell.

METHODPC-12 cells were cultured with nerve growing factors (NGF), and pretreated with Sin at various concentrations (0, 3 x 10(-6), 30 x 10(-6), 150 x 10(-6) mol x L(-1)) for 2 hours, then with or without stimulation of lipopolysaccharide (LPS). The proliferation activity of PC-12 cells was determined by 3H-TdR incorporation, and the production of PGE2 in culture supernatants of PC-12 cells was detected with competitive ELISA. Expression of COX-2 mRNA in PC-12 cells was analyzed by semi-quantitative RT-PCR, and expression of COX-2 protein was estimated by Western blot method and cellular enzyme immunoassay. Nuclear factor-kappa B (NF-kappaB) activity in whole-cell extract of PC-12 cells was also measured by an ELISA-based method.

RESULTThe data showed that Sin down-regulated the expression of COX-2 mRNA and protein, and reduced the production of PGE2 in the LPS-stimulated PC-12 cells which correlated with Sin's concentrations positively. In addition, NF-kappaB activity in LPS-stimulated cells was suppressed significantly by Sin. No inhibition of proliferation of PC-12 cells due to Sin treatment was observed.

CONCLUSIONSin mediates the down-regulation of expression of COX-2 and production of induced PGE2 in PC-12 cells by suppressing the activity of NF-kappaB.

Animals ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; Dinoprostone ; biosynthesis ; Down-Regulation ; Lipopolysaccharides ; Morphinans ; isolation & purification ; pharmacology ; NF-kappa B ; metabolism ; PC12 Cells ; Plants, Medicinal ; chemistry ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Sinomenium ; chemistry

OBJECTIVETo observe in vitro the effect of Sinomenine, a pure alkaloid extracted from the chinese medical plant Sinomenium acutum on the activity of cyclooxygenase (COX-1 and COX-2) and the expression of COX-1 and COX-2 mRNA.

METHODMononuclear leukocytes were obtained from healthy adults. Isolated mononuclear leucocytes from human peripheral blood (PBMC) were incubated (1 x 10(6).mL-1) with or without sinomenine (or indomethacin), after incubated for 24 hours at 37 degrees C with 5% CO2; the media were assayed for the PGE2 by radioimmunoassay (RIA). LPS was used to stimulate the monocytes at a concentration of 5 micrograms.mL-1. And by RT-PCR, both COX-1 and COX-2 mRNAs were detected in Mononuclear leukocytes after incubation for different hours with drug (sinomenine or indomethacin) or not.

RESULTLPS (stimulated) induced the production of PGE2 in PBMC increasing with high expression of COX-2 mRNA; sinomenine reduced PGE2 production in LPS stimulated human monocytes more than in non-stimulated human monocytes. In comparative experiments, indomethacin, a non selective COX inhibitor, reduced the production of PGE2 equally in both states. Meanwhile, neither sinomenine(0.1-1 mmol.L-1) nor indomethacin(0.5-10 mumol.L-1) inhibited the expression of both COX-1 and COX-2 mRNAs by RT-PCR with beta-actin as reference.

CONCLUSIONIn contrast with indomethacin, Sinomenine shows a preferential inhibitory effect on COX-2 over COX-1, These results suggest that Sinomenine is a selective COX-2 inhibitor, which may be directly related to suppressing cyclooxygenase activity.

Adult ; Cyclooxygenase 1 ; Cyclooxygenase 2 ; Dinoprostone ; blood ; Humans ; Isoenzymes ; biosynthesis ; Leukocytes, Mononuclear ; enzymology ; Membrane Proteins ; Morphinans ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; metabolism ; RNA, Messenger ; biosynthesis ; Sinomenium ; chemistry

OBJECTIVETo establish a fingerprint analysis method of Qingfengtong capsule by HPLC.

METHODThe samples were extracted with 70% ethanol in an ultrasonic bath. The extracts were analyzed at 35 degrees C on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) with 0.1 mol x L(-1) potassium dihydrogen phosphate water-solution as mobile phase A, and methanol as mobile phase B. The analysis followed a linear gradient elution program. Initial condition of the mobile phase was 10% B for 2 minutes; then changed to 90% B in 40 minutes. The flow rate was kept at 1.0 ml x min(-1) and the detector wavelength was 262 nm.

RESULTThe main peaks in fingerprint chromatogram of Qingfengtong capsules were separated fairly well. The results of method validation meet the requirements for the fingerprints.

CONCLUSIONThe established method can be used for the quality control of Qingfengtong capsules.

Capsules ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; standards ; Epimedium ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Sinomenium ; chemistry