ABSTRACT
In order to discuss the "entropy weight method" for weighting various indicators in the comprehensive evaluation of Angelicae Sinensis Radix slices(ASR), the quality of ASR was comprehensively evaluated by entropy weight-based gray systematic theory and cluster analysis. In this study, the contents of ferulic acid, volatile oil, polysaccharide, alcohol extract, water extract, moisture, total ash and acid-insoluble ash in 44 batches of ASR from different sources were determined. The entropy weight method was used for objective weighting. With relative correlation(r_i) as a measure, a multi-index comprehensive evaluation model was constructed for the quality of ASR. The results showed that the relative correlation value of 44 batches of ASR ranged from 0.301 9 to 0.662 9. There were certain differences in the quality of ASR from different sources. The ASR S1-S8, traceable and standardized in processing techno-logy, showed a high relative correlation degree and high quality ranking, indicating that the implementation of systemic management of the production chain of Chinese herbal pieces was beneficial to the quality control of ASR. The quality evaluation results of 44 batches of ASR were consistent with those of traditional geo-authentic habitats for ASR and the mainstream varieties of ASR on market, and basically consistent with the results of cluster analysis. This study suggests that the gray systematic theory based on the entropy weighting method can be used for the quality evaluation of ASR. The objective weighting of the entropy weight method improves the reliability of the gray correlation method and the scientificity of ASR quality evaluation.
Subject(s)
Angelica sinensis , Drugs, Chinese Herbal , Entropy , Oils, Volatile , Plant Roots , Reproducibility of ResultsABSTRACT
Objective: The solubilization of 4 compounds(Euphorbia factor L1,L2,L3 and L8),caused by mixed micelles self-assembled from fatty oil of Euphorbiae Semen and bile salt of intestinal juice,was researched in the simulated human intestinal environment. Method: The mixed micelles were prepared with different amounts of fatty oil of Euphorbiae Semen.The transmission electron microscope(TEM) was used to observe the morphology of the micelles.Particle size detector was used to determine the particle size and Zeta potential.HPLC was used to assay the solubility of these 4 compounds.The variation tendency of the total dissolution of these 4 compounds with the change of standing time was observed. Result: Particle size of the mixed micelles was uniform and its morphology was spherical.The absolute values of Zeta potential were less than 20 mV.When the amount of sodium deoxycholate was fixed 4.96 g·L-1,the solubility of these 4 compounds with the concentration of fatty oil at 0.1-4 g·L-1 were significantly greater than that at the dosage of 0 g·L-1.The solubility of these 4 compounds in the micelles formed by fatty oil was 1.3 to 4 times as much as the micelles without fatty oil.The micelles was stable for 36 h. Conclusion: The micelles self-assembled from fatty oil of Euphorbiae Semen and bile salt of intestinal juice,have significant solubilization effect on Euphorbia factor L1,L2,L3 and L8.This research can lay the foundation for clarifying the detoxification mechanism of removing fatty oil and making frostlike powder from the perspective of pharmaceutics.
ABSTRACT
The present research was designed to study expression of AQP2, AQP4 and AQP8 in mouse intestines induced by unprocessed and processed Euphorbia lathyris. KM mice were given by different dose lavage of unprocessed and processed Euphorbia lathyris, Euphorbia factor L1, Euphorbia factor L2, Euphorbia factor L3. Samples of mouse intestine were collected for protein levels of AQP2, AQP 4 and AQP 8 which were assessed by immunohistochemical staining and mRNA expression of AQP2, AQP 4 and AQP 8 which were quantified by Real Time-PCR. Comparing to the normal control group, the protein levels of AQP2, AQP 4 and AQP 8 were significantly decreased [P<0.05]by Semen Euphorbiae group and Semen Euphorbiae Pulveratum group [unprocessed and processed Euphorbia lathyris] induced. Protein expression of AQP2, AQP 4 and AQP 8 in the Euphorbia factor L1, Euphorbia factor L2 and Euphorbia factor L3 group were not significantly lower than normal control group. There had no differences on the levels of AQP2 and AQP 8 mRNA expressions between the high-dose group of semen Euphorbiae group, semen Euphorbiae Pulveratum group and positive control group, while significantly lower than normal control group [P<0.05]. Expression of AQP4 mRNA in the Semen Euphorbiae group and Semen Euphorbiae Pulveratum group has not significantly decreased. But levels of AQP2, AQP 4 and AQP 8 mRNA in the Euphorbia factor L1 group had no significant differences in normal control group and positive control group. These findings suggest that semen Euphorbiae could regulate expression of AQP2, AQP 4 and AQP 8 protein and mRNA, which may be the possible one reason of semen Euphorbiae induces diarrhea. The semen Euphorbiae group has more significant effects on the levels of AQP2, AQP 4 and AQP 8 protein and mRNA than semen Euphorbiae Pulveratum group, which may be one of the mechanisms of processing attenuation
ABSTRACT
A new method was developed for the chromatographic fingerprint analysis of Toosendan Fructus by HPLC coupled with the charged aerosol detector (CAD) in the present study. Samples were well separated on an Agilent ZOBAX SB C18 column (4.6 mm×250 mm, 5 μm) by gradient elution using acetonitrile and water containing 0.1% formic acid (v/v) at the flow rate of 1.0 mL·min-1. The column temperature was 30℃ and the injection volume was 5 μL. The nitrogen inlet pressure of the charged aerosol detector (CAD) was 35 psi, and the nebulizer chamber temperature was 35℃. In addition, the method of the chromatographic fingerprints combined with multivariate statistical analysis was effective and reasonable lead to an accurate classification of 20 batches of samples from different locations. The results showed that 28 common peaks were observed in the fingerprint and the samples were classified into three clusters. The established method was well validated, and showed high precision, good repeatability, and satisfactory stability. It may serve in the quality control and evaluation of Toosendan Fructus.
ABSTRACT
Study on the standardization of Chinese materia medica is an important action for modernization and globalization for traditional Chinese medicine. Standardization on the processing of Chinese herbal pieces is an important part in the study on standardization of Chinese materia medica, so it is of great significance to establish the technical processing standards of Angelicae Sinensis Radix pieces for improving its quality. In this study, single factor experiment was designed to optimize the softening, cutting and drying processes of Angelicae Sinensis Radix. With ferulic acid, Angelicae Sinensis Radix polysaccharide, volatile oil and extracts (water and ethanol) content as the quality index, the effects of different softening, cutting and drying processes on the contents of the five components in Angelicae Sinensis Radix were analyzed, and the normalized distance evaluation method was used to analyze the experimental data. The results showed that the content of five components in Angelicae Sinensis Radix was affected by different softening methods and drying temperature, but the thickness of slice had little effect on the content. The best preparation process for Angelicae Sinensis Radix was as follows: Non-medicinal parts were removed; mildewed and rot as well as moth-eaten parts were removed; washed by the flowing drinking water; stacked in the drug pool; moistening method was used for softening, where 125 mL water was sprayed for every 1 kg of herbs every 2.5 h; upper part of herbs covered with clean and moist cotton, and cut into thin slices (1-2 mm) after 15 h moistening until appropriate softness, with disk thickness of 1-2 cm, then received blast drying for 6 h at 55 ℃, and turned over for 2 times during the drying.
ABSTRACT
Notopterol, isoimperatorin, volatile oil and extract （water and ethanol) were used as the research objects in this study to investigate the effects of different softening method, slice thickness and drying methods on the quality of Notopterygii Rhizoma et Radix slices, and the experimental data were analyzed by homogeneous distance evaluation method. The results showed that different softening, cutting and drying processes could affect the content of five components in Notopterygii Rhizoma et Radix incisum. The best processing technology of Notopterygii Rhizoma et Radix slices was as follows: non-medicinal parts were removed; mildewed and rot as well as moth-eaten parts were removed; washed by the flowing drinking water; stacked in the drug pool; moistening method was used for softening, where 1/8 volume of water was sprayed for every 1 kg of herbs every 2 h; upper part of herbs covered with clean and moist cotton, and cut into thick slices (2-4 mm) after 12 h moistening until appropriate softness, then received blast drying for 4 h at 50 ℃, and turned over for 2 times during the drying. The process is practical and provides the experimental basis for the standardization of the processing of Notopterygii Rhizoma et Radix, with great significance to improve the quality of Notopterygii Rhizoma et Radix slices.
ABSTRACT
The mechanism underlying the modulatory effect of substance P (SP) on GABA-activated response in rat dorsal root ganglion (DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clamp technique was used to record GABA-activated current and sharp electrode intracellular recording technique was used to record GABA-induced membrane depolarization. Application of GABA (1-1000 μmol/L) induced an inward current in a concentration-dependent manner in 114 out of 127 DRG neurons (89.8 %) examined with whole-cell patch-clamp recordings. Bath application of GABA (1-1000 μmol/L) evoked a depolarizing response in 236 out of 257 (91.8%) DRG neurons examined with intracellular recordings. Application of SP (0.001-1 μmol/L) suppressed the GABA-activated inward current and membrane depolarization. The inhibitory effects were concentration-dependent and could be blocked by the selective neurokinin 1 (NK1) receptors antagonist spantide but not by L659187 and SR142801 (1 μmol/L, n=7), selective antagonists of NK2 and NK3. The inhibitory effect of SP was significantly reduced by the calcium chelator BAPTA-AM, phospholipase C (PLC) inhibitor U73122, and PKC inhibitor chelerythrine, respectively. The PKA inhibitor H-89 did not affect the SP effect. Remarkably, the inhibitory effect of SP on GABA-activated current was nearly completely removed by a selective PKCε inhibitor epilon-V1-2 but not by safingol and LY333531, selective inhibitors of PKCα and PKCβ. Our results suggest that NK1 receptor mediates SP-induced inhibition of GABA-activated current and membrane depolarization by activating intracellular PLC-Ca²⁺-PKCε cascade. SP might regulate the excitability of peripheral nociceptors through inhibition of the "pre-synaptic inhibition" evoked by GABA, which may explain its role in pain and neurogenic inflammation.
Subject(s)
Animals , Female , Male , Rats , Ganglia, Spinal , Physiology , Patch-Clamp Techniques , Protein Kinase C-epsilon , Metabolism , Rats, Sprague-Dawley , Receptors, GABA-A , Physiology , Signal Transduction , Substance P , PhysiologyABSTRACT
The mechanism underlying the modulatory effect of substance P (SP) on GABA-activated response in rat dorsal root ganglion (DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clamp technique was used to record GABA-activated current and sharp electrode intracellular recording technique was used to record GABA-induced membrane depolarization. Application of GABA (1-1000 μmol/L) induced an inward current in a concentration-dependent manner in 114 out of 127 DRG neurons (89.8 %) examined with whole-cell patch-clamp recordings. Bath application of GABA (1-1000 μmol/L) evoked a depolarizing response in 236 out of 257 (91.8%) DRG neurons examined with intracellular recordings. Application of SP (0.001-1 μmol/L) suppressed the GABA-activated inward current and membrane depolarization. The inhibitory effects were concentration-dependent and could be blocked by the selective neurokinin 1 (NK1) receptors antagonist spantide but not by L659187 and SR142801 (1 μmol/L, n=7), selective antagonists of NK2 and NK3. The inhibitory effect of SP was significantly reduced by the calcium chelator BAPTA-AM, phospholipase C (PLC) inhibitor U73122, and PKC inhibitor chelerythrine, respectively. The PKA inhibitor H-89 did not affect the SP effect. Remarkably, the inhibitory effect of SP on GABA-activated current was nearly completely removed by a selective PKCε inhibitor epilon-V1-2 but not by safingol and LY333531, selective inhibitors of PKCα and PKCβ. Our results suggest that NK1 receptor mediates SP-induced inhibition of GABA-activated current and membrane depolarization by activating intracellular PLC-Ca(2+)-PKCε cascade. SP might regulate the excitability of peripheral nociceptors through inhibition of the "pre-synaptic inhibition" evoked by GABA, which may explain its role in pain and neurogenic inflammation.
ABSTRACT
<p><b>OBJECTIVE</b>To discuss the optimum extraction process of compound Nanxing pain-relieving cataplasm through orthogonal design and pharmacodynamic experiment</p><p><b>METHOD</b>The orthogonal experiment method was adopted to optimize the ethanol extraction process with Angelica dahurica, Ligusticum chuanxiong and other herbs. The anti-inflammatory and analgesic effects of extracts from volatile oil in such herbs as syzygium aromaticum with different extraction processes were compared by tail pain tenderness test and food-pad swelling in mice, in order to optimize the extraction process of extracts from volatile oil in such herbs as syzygium aromaticum.</p><p><b>RESULT</b>The optimum extraction process of A. dahurica, L. chuanxiong and other herbs for compound Nanxing pain-relieving cataplasm were as follows: adding 8-fold amount of 70% alcohol, extracting for 2 times with 1.5 h each time. The 95% ethnol extracts of syzygium aromaticum and other herbs had more effect in the increasing the threshold of pain and the inhibition of toe swelling of mice than volatile oil obtained from steam distillation as well as volatile oil and water decoction obtained from steam distillation.</p><p><b>CONCLUSION</b>The method is simple and reliable that it can provide technical reference for the development of modern preparations of compound Nanxing pain-relieving cataplasm.</p>
Subject(s)
Animals , Mice , Analgesics , Chemistry , Pharmacology , Therapeutic Uses , Chemical Fractionation , Methods , Chemistry, Pharmaceutical , Methods , Drugs, Chinese Herbal , Chemistry , Pharmacology , Therapeutic Uses , Ethanol , Chemistry , Oils, Volatile , Chemistry , Pain , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To study transdermal absorption characteristics of eugenol in compound Nanxing pain-relieving cataplasm, and discuss the effect of gaultherolin on the transdermal absorption of the cataplasm.</p><p><b>METHOD</b>The improved franz diffusing cell was adopted with hairless mice skins as transdermal carriers. The content of eugenol in receptor liquid, skins and cataplasm were analyzed by HPLC and compared with the cataplasm without gaultherolin.</p><p><b>RESULT</b>The penetration rates of eugenol of cataplasms with and without gaultherolin were 13.18 and 9.58 microg x cm(-2) x h(-1), with the retention amount in skins of (185.02 +/- 19.23) and (160.23 +/- 16.54) microg x g(-1) and the retention amount in cataplasms was (1.96 +/- 0.12) and (1.71 +/- 0.15) mg, respectively.</p><p><b>CONCLUSION</b>Eugenol in compound Nanxing pain-relieving cataplasm has good pereutaoeous permeation. Gaultherolin in the cataplasm prescription can promote the absorption of eugenol.</p>
Subject(s)
Animals , Mice , Analgesics , Chemistry , Metabolism , Therapeutic Uses , Chemistry, Pharmaceutical , Methods , Drugs, Chinese Herbal , Chemistry , Metabolism , Therapeutic Uses , Pain , Drug Therapy , Salicylates , Chemistry , Skin , Metabolism , Skin Absorption , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To prepare Kushen-Dilong nanoemulsion and nanoemuls-ion gel, and investigate its content, physical and chemical properties. Their transdermal properties in vitro were studied as well.</p><p><b>METHOD</b>IPM acted as oil phase, EL35 as surfactant, EtOH as cosurfactant, Pheretima aqueous solution was added dropwise to the oil phase to prepare Kushen-Dilong nanoemulsion at room temperature using magnetic stirring. HPLC was used to determine the content of matrine and oxymatrine in the nanoemulsion. Transmission electron microscopy and laser particle size analyzer was used to determine the shape and size of the nanoemulsion. NP700 was used as substrate to prepare Kushen-Dilong nanoemulsion gel. Franz diffusion cell was used for the nanoemulsion and gel transdermal characteristics in vitro.</p><p><b>RESULT</b>The Kushen-Dilong nanoemulsion was O/W nanoemulsion, its uniform particle size was 20.6 nm with roundness appearance and stable content. The steady-state permeation rate of Kushen-Dilong nanoemulsion, nanoemulsion gel, saturated aqueous solution, hydro gel were 0.1484, 0.1183, 0.0306, 0.0321 mg x cm(-2) x h(-1), respectively.</p><p><b>CONCLUSION</b>The 24 h cumulative infiltration and infiltration rate of Kushen-Dilong nanoemulsion and nanoemulsion gel were better than the saturated aqueous solution and hydro gel, which could provide a new dosage form for Kushen-Dilong transdermal drug delivery.</p>
Subject(s)
Animals , Rats , Chemistry, Pharmaceutical , Drug Carriers , Chemistry , Drugs, Chinese Herbal , Chemistry , Metabolism , Emulsions , Gels , Nanostructures , Chemistry , Rats, Sprague-Dawley , Skin , Metabolism , Skin AbsorptionABSTRACT
The research aimed at investigating the physicochemical properties, stability and skin penetration in vitro of total alkaloids of Sophora flavescens nanoemulsion. Prepare total alkaloids of S. flavescens nanoemulsion and detect the determination of matrine and oxymatrine in the nanoemulsion using HPLC method. Transmission electron microscopy and laser particle size analyzer were utilized to detect the shape and size of the nanoemulsion respectively. And also the stability of nanoemulsion was studied under the conditions of low temperature (4 degrees C), normal temperature (25 degrees C) and high temperature (60 degrees C). Franz diffusion cell was used to research the transdermal absorption of nanoemulsion in vitro. The results found that the nanoemulsion we prepared presented appearance of rounded, uniform; its average diameter was (15.55 +/- 2.24) nm, and particle size distribution value was 0. 161; the appearance, diameter and percentage determination of total alkaloids of S. flavescens had no variations after 15 d under 4, 25, 60 degrees C respectively. The steady-state permeation rate was 4.564 1 microg x cm(-2) x h(-1), 24 h cumulative amount of penetration was 110.7 microg x cm(-2), which was 1.86 fold of 24 h cumulative amount of aqueous solution (59.41 microg x cm(-2)). All the results demonstrated total alkaloids of S. flavescens nanoemulsion had good permeability, and could provide a new preparation for its clinical application.
Subject(s)
Animals , Male , Rats , Alkaloids , Chemistry , Metabolism , Chemical Phenomena , Drug Carriers , Chemistry , Emulsions , Nanostructures , Chemistry , Rats, Sprague-Dawley , Skin Absorption , Sophora , ChemistryABSTRACT
Chloride channels have been identified in vascular smooth muscle cells (SMCs). It has been shown that these channels are involved in myogenic tone regulation and neuromuscular transmission in various vascular beds. However, whether the chloride channels are responsible for the formation of excitatory junction potentials (EJPs) of SMCs in the spiral modiolar artery (SMA) remains unelucidated. In the present study, the effects of chloride channel blockers (niflumic acid, NFA; indanyloxyacetic acid 94, IAA-94; disodium 4, 4'-diisothiocyanatostilbene-2, 2'-disulfonate, DIDS) on EJP were explored in guinea pigs, using intracellular recording techniques on acutely isolated SMA. It was found that EJP was evoked in the majority of the SMCs (75%, n=49) with an adequate electronic stimulation. The amplitude of the EJP was partially blocked (30% approximately 80%) by combined application of alpha(1) receptor antagonist (prazosin) and alpha(2) receptor antagonist (idazoxan) at concentration of up to 1 micromol/L, and P(2x) receptor antagonist (PPADS, 10 approximately 100 micromol/L). NFA (100 micromol/L) could further inhibit the residual EJP in the presence of alpha(1), alpha(2)-adrenergic and P(2x) receptor antagonists. IAA-94 or DIDS not only inhibited the amplitude but also shortened the duration of EJP. Decrease of extracellular chloride concentration from 135.6 mmol/L to 60 mmol/L would enhance EJP. Moreover, IAA-94 (100 micromol/L) and DIDS (200 mumol/L) could reverse the enhancement of EJP by low extracellular Cl(-). NFA (100 micromol/L) could also block the residual depolarizations evoked by norepinephrine (NE, 1 approximately 50 micromol/L). Based on these results, it is inferred that NE could activate a novel adrenoceptor to open the chloride channel on the membrane of the SMCs, leading to a transmembrane Cl(-) current. This current is involved, at least partially, in the formation of EJP.
Subject(s)
Animals , Female , Male , Adrenergic alpha-Antagonists , Pharmacology , Arteries , Physiology , Chloride Channels , Cochlea , Excitatory Postsynaptic Potentials , Guinea Pigs , Muscle, Smooth, Vascular , Cell Biology , Physiology , Myocytes, Smooth Muscle , Physiology , Norepinephrine , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To study the effects of different penetration enhancers on the transcutaneous permeability of lappaconitine gel in vitro and therefore to screen out the effective accelerator to enhance the permeation rate of lappaconitine.</p><p><b>METHOD</b>Using improved Franz-type diffusion cell and excised big mouse skin in vitro as transdermal barrier, the kinetics parameters 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 (Azone), propylene glycol (PG), oleic acid (OA) and lauryl alcohol (LA), was investigated when used either uniquely or combinatively each other at random.</p><p><b>RESULT</b>When used combinatively, Azone + PG, LA + PG, OA + PG can enhance the cumulative permeation quantity, OA + PG was the best one among them.</p><p><b>CONCLUSION</b>The selection of the best penetration enhancers provided reference for lappaconitine transdermal delivery.</p>
Subject(s)
Animals , Female , Male , Rats , Aconitine , Pharmacokinetics , Administration, Cutaneous , Analgesics, Non-Narcotic , Pharmacokinetics , Azepines , Pharmacology , Dodecanol , Pharmacology , Drug Combinations , Drug Synergism , In Vitro Techniques , Oleic Acid , Pharmacology , Propylene Glycol , Pharmacology , Rats, Sprague-Dawley , Skin AbsorptionABSTRACT
The crude toxin was extracted from hypha and culture solution of Phyllosticta commelimecola through three different polarity solvent: benzinum, puncificatum ethyl acetate and chloroform. The result indicated that the toxin secreted by Phyllosticta commelimecola not only was in hypha but also in culture solution and the extracting effect of ethyl acetate was the best. The soybean median and PSK media can be respectively used as solid and liquid culture media to produce toxin and grow mycelium. The optimal cultural conditions for producing toxin were temperature 32℃,cultured period 14d, cultured ways shaking of 150r/min.