Dipsaci Radix is one of the commonly used Chinese medicinal materials in China, with a long history. It has the medicinal activities of nourishing liver and kidney, recovering from broken sinews, and treating bone fracture. Triterpenoid saponins are the main functional ingredients of Dipsacus asper. β-Amyrin synthases(β-AS) as a superfamily of oxidosqualene cyclases(OSCs) can catalyze the construction of the skeleton structure of oleanane-type triterpenoid saponins. There are only a few studies about the β-AS in D. asper, and the catalytic mechanism of this enzyme remains to be explored. To enrich the information of β-AS, according to the transcriptome sequencing results, we cloned DaWβ-AS gene from D. asper into a specific vector for heterologous expression in Escherichia coli. In the meantime, real-time PCR was performed to analyze the relative expression of DaWβ-AS in four different tissues of D. asper. The results of RT-qPCR showed DaWβ-AS had the highest expression level in leaves. Bioinformatics results indicated that DaWβ-AS had a conserved domain of PLN03012 superfamily, belonging to the cl31551 superfamily. There was no transmembrane domain or signal peptide in DaWβ-AS. This study provides a scientific basis for revealing the biological pathways of triterpenoid saponins in D. asper, which will facilitate the biosynthesis of the associated saponins and afford reference for the cultivation and development of high-quality resources of D. asper.
Cloning, Molecular ; Computational Biology ; Dipsacaceae/chemistry* ; Intramolecular Transferases ; Protein Sorting Signals ; Saponins/chemistry* ; Triterpenes/chemistry*

Through the records of herbal and market investigations,the samples collected from different market and origin were analyzed,and the appearance character indexes were determined in order to revise the commodity specification and grade standard of Dipsaci Radix combined with production practice. This is also to analyze association of the appearance with quality different of intrinsic components. The investigation results indicated the root's long and thick was better,and atrovirens of fracture surface,there are most of the market is dominated by the ungraded goods. Through principal component analysis,variance analysis and cluster analysis,combining with the feasibility of actual operation and herbal record,the length,middle diameter and phloem color were filtrated and the specification was divided into 2 ranks: the selection and ungraded goods,and the grade was divided into 2 ranks for the selection goods: big,small selected goods. Moreover,there were no significant correlation between the appearance and the intrinsic quality index of Dipsaci Radix,the content of extractum were significantly positive correlation with the moisture,but had extremely negative correlation with the total ash. Multiple comparisons indicated that the content of the moisture,extractum and asperosaponin VI of the selected goods were higher than the ungraded goods,while the total ash content were lower,and they hasn't significant difference. The commodity specification and grade standard of Dipsaci Radix as a basis provide commodity specification and grade standard of communities and standardizing market trade order.
Cluster Analysis ; Dipsacaceae ; chemistry ; Drugs, Chinese Herbal ; standards ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Principal Component Analysis

To investigate the genetic diversity among wild Dipsacus asperoides in China, 66 germplasmic resources of D. asperoides were analyzed by Start Codon Targeted Polymorphism (SCoT) molecular markers. Genetic distance was calculated by TREECONW software and the systematic diagram of genetic relationship was clustered by UPGMA method. The results showed that the totals of 181 bands were detected using 20 primers , among which 109 were polymorphic bands. The average percentage of polymorphic bands was 60.13%. Genetic distance changed from 0.030 6 to 0.181 4. The clustering results showed that there was no significant correlation between the classification of the wild D. asperoides and their geographical origin. The relatively high genetic diversity of D. asperoides provides the basis for breeding new varieties.
China ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Dipsacaceae ; chemistry ; classification ; genetics ; Genetic Variation ; Phylogeny ; Polymorphism, Genetic

Twenty-two compounds were isolated from the flowers of Scabiosa tschilliensis. Their structures were identified by spectroscopic methods as octacosanol (1), stearic acid (2), β-sitosterol (3), oleanolic acid (4), apigenin (5), luteolin (6), daucosterol (7), kaempferol-3-O-β-D-6-O-(p-hydroxycinnamoyl) -glucopyranoside (8), kaempferol-3-O-β-D- (3, 6-di-p-(hydroxycinnamoyl) -glucopyranoside (9), apigenin-7-O-β-D-glucopyranoside (10), luteolin-4'-O-β-D-glucopyranoside (11), apigenin-7-O-rutinoside (12), luteolin-7-O-β-D-glucopyranoside (13), apigenin-4'-O-β-D-glucopyranoside (14), caffeic acid methyl ester (15), loganin (16), adenosine (17), luteolin-6-C-β-D-glycopyranosyl (18), sweroside (19), sylvestrosides I (20), sylvestrosides II (21), urceolide (22). Among them, compounds 1, 2, 7-9, 12, 15, 17-18, 20-22 were isolated from the genus Scabiosa for the first time, and compounds 1-4, 6-9, 11-12, 14-22 were isolated from this plant for the first time. 13C-NMR data of 22 were reported for the first time.
Dipsacaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Flowers ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo Study the effect of anti-aging and its mechanism of total saponins of Wu-He Dipsacus asper on skin of mice-aging model.

METHODSForty-eight mice were randomly divided into blank control group, model group, low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group and positive control group( n = 8) . The mouse model of skin aging was established by nape subcutaneous injection of 5% D-galactose (0.025 mL/(g · d)), the mouse of low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group were administered with total saponins of Wu-He Dipsacus asper (50 ml/(kg · d), 100 mL/(kg · d), 200 mL/(kg · d)), the mice of the positive control group were administered with vitamin E(50 mg/(kg · d)) for 42 d. The content of hydroxyproline (HYP) and lipofuscin (LF) were measured in skin of each group mice, the activity of catalase (CAT) glutathione peroxidase ( GSH-Px) superoxide dismutase (SOD) and the content of malondi- aldehyde (MDA) were determined in serum and skin of each group mice.

RESULTSCompared with blank control group, the content of HYP decreased significantly and the content of LF increased significantly in skin, the activities of CAT, GSH-Px and SOD decreased significantly and the content of MDA increased significantly in serum and skin of model group; Compared with model group, the content of HYP increased significantly and the content of LF decreased significantly in skin, the activities of CAT, GSH-Px and SOD enhanced significantly and the con- tent of MDA decreased significantly in serum and skin of low-Dipsacus group, medium-Dipsacus group, high-Dipsacus group and positive control group; Compared with low-Dipsacus group, the content of HYP increased significantly and the content of LF decreased significantly in skin, the activities of CAT, GSH-Px and SOD enhanced significantly and the content of MDA decreased significantly in serum and skin of high-Dipsacus group and positive control group; The activity of SOD in serum and skin had a significant positive correlation with the content of HYP, and a significant negative correlation with LF in skin.

CONCLUSIONTotal saponins of Wu-He Dipsacus asper have obvious effect of anti-agng on skin of mouse-aging model , its mechanism is closely related to oxidative damage.

Animals ; Dipsacaceae ; chemistry ; Disease Models, Animal ; Mice ; Oxidative Stress ; Saponins ; pharmacology ; Skin Aging ; drug effects

OBJECTIVETo study the effects of Dipsacus total saponins on the ability of learning and memory and its mechanism of action.

METHODSForty rats were randomly divided into blank control group, model group, Dipsacus group and positive control group (n = 10), general situation of rats were observed, the ability of learning and memory of rats was tested by Square water maze, the activities of acetylcholinesterase (AChE)and choline acetyltransferase (ChAT) of hippocampus in rats were measured using double antibody sandwich method.

RESULTSDuring the period of treatment, general situation had no obvious change in model group, but general situation and the ability of activity were gradually improved in Dipsacus group and positive control group. Compared with blank control group, the swimming time was obviously prolonged and the number of mistakes was obviously increased at different time, the activity of AChE was significantly enhanced and the activity of ChAT was significantly decreased in model group. Compared with model group, the swimming time was obviously shortened and the number of mistakes was obviously reduced at different time, the activities of AChE were significantly decreased and the activities of ChAT were significantly enhanced in Dipsacus group and positive control group; Compared with positive control group, the swimming time and the number of mistakes at different time and the activities of AChE and ChAT had no significant difference in Dipsacus group.

CONCLUSIONDipsacus total saponins can improve the ability of learning and memory in Alzheimer' s disease(AD) rats, its mechanism of 'action may be related to regulating ACh metabolism of hippocampus.

Acetylcholine ; metabolism ; Acetylcholinesterase ; metabolism ; Alzheimer Disease ; drug therapy ; physiopathology ; Animals ; Choline O-Acetyltransferase ; metabolism ; Dipsacaceae ; chemistry ; Disease Models, Animal ; Hippocampus ; drug effects ; Learning ; drug effects ; Memory ; drug effects ; Rats ; Saponins ; pharmacology

OBJECTIVETo study the effect of Wu-He Dipsacus asper (WHDA), Traditional Chinese Medicine, injection on mice-aging model induced by D-galactose.

METHODSForty-eight Kunming mice (24 male and 24 female) were randomly divided into control group, model group, positive control group, 7.2 g/kg WHDA group, 3.6 g/kg WHDA group and 1.8 g/kg WHDA group with eight in each group. The model was induced through injecting D-galactose into peritoneal cavity and Morris water maze was used to detect the learning and cognitive ability of mice. The skin hydroxyproline, brain tissue malondialdehyde (MDA), lipofuscin (LP), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels of mice were detected; the IL-2 and IL-6 levels in serum of mice were detected by using double antibody sandwich ELISA method.

RESULTSEach WHDA group was significantly reduced in latency period compared with the model group during Morris water maze test (P < 0.05) and the number of mice in model group through the platform was less than other mice in each group (P < 0.05). The levels of MAD and LP of the control group and each WHDA group were less than model group in the detection of heart, brain tissue oxidation index (SOD, MAD, LP and GSH-Px, P < 0.05). The activity of SOD and GSH-Px in the control group and each WHDA group was significantly higher than that in the model group (P < 0.05). The skin hydroxyproline content of mice which had been injected with D-galactose was significantly lower than that in the control group (P < 0.05) and the skin hydroxyproline content of mice of WHDA group was significantly higher than that in the model group (P < 0.05). The IL-2, IL-6 levels in serum of mice in WHDA group were significantly higher than those in the control group and the model group (P < 0.05) and the IL-2, IL-6 levels in serum of mice in the model group were lower than those in the control group (P < 0.05).

CONCLUSIONThe effective constituents of WHDA have a variety of biological activity which can have a good effect on anti-aging by different ways, improving learning and memory function, eliminating free radicals antioxidant, and enhancing the body immunity and other aspects.

Aging ; drug effects ; physiology ; Animals ; Brain ; drug effects ; metabolism ; Dipsacaceae ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Galactose ; toxicity ; Glutathione Peroxidase ; metabolism ; Hydroxyproline ; metabolism ; Interleukins ; blood ; Learning ; drug effects ; Lipofuscin ; metabolism ; Male ; Malondialdehyde ; metabolism ; Memory ; drug effects ; Mice ; Skin ; drug effects ; metabolism ; Superoxide Dismutase ; metabolism

In order to discriminate the crude and sweated Dipsaci Radix correctly and rapidly, the crude and sweated Dipsaci Radix were scanned by the NIR spectrometer, and an identifying model was developed by near infrared spectroscopy combined with principal component-Mahalanobis distance pattern recognition method. The pretreated spectra data of 129 crude samples and 86 sweated ones were analyzed through principal component analysis (PCA). The identifying model was developed by choosing the spectrum for 9 881.46-4 119.20 cm(-1) and "SNV + spectrum + S-G" to the original spectral preprocessing with 14 principal components, and then was verified by prediction set, identifying with 100% accuracy. The rapid identification model of the crude and sweated Dipsaci Radix by NIR is feasible and efficient, and could be used as an assistant means for identifying the crude and sweated Dipsaci Radix.
Chemistry, Pharmaceutical ; methods ; Dipsacaceae ; chemistry ; Plant Roots ; chemistry ; Principal Component Analysis ; methods ; Quality Control ; Spectroscopy, Near-Infrared ; methods

AIM: To establish and apply a new LC/MS/MS method for the simultaneous, quantitative determination of six ingredients, aucubin (AU), geniposide (GP), geniposidic acid (GPA), pinoresinol diglucoside (PDG), secologanin (SLG), and loganin (LG) in single and combined extracts of Eucommia ulmoides and Dipsacus asperoides. METHOD: Using the LC/MS/MS-ESI(-)-MRM mode to detect the six compounds, chromatographic separation was achieved on an Agilent Eclipse plus C18 column, and the mobile phase consisted of solvent A (CH3CN) and solvent B (H2O containing 0.01% CH3COOH V/V). RESULTS: This method was successfully applied to quantify the six compounds in rat plasma after oral administration, and showed good precision, accuracy, reproducibility, and linear regression (r(2)>0.99). CONCLUSION The results showed that following the use of the two medicinal plants, for AU and GP, the values of Cmax markedly increased, and the values of cmax markedly decreased. It was found that the compatibility of the medicinal plants might affect their pharmacokinetic properties of their constituents.
Animals ; Chromatography, High Pressure Liquid ; methods ; Dipsacaceae ; chemistry ; Drug Combinations ; Eucommiaceae ; chemistry ; Female ; Iridoid Glucosides ; blood ; pharmacokinetics ; Iridoids ; blood ; pharmacokinetics ; Lignans ; blood ; pharmacokinetics ; Plant Extracts ; blood ; chemistry ; pharmacokinetics ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Model organism zebrafish was used to study metabolism of asperosaponin VI from Dipsacus asper Wall. ex Henry for the first time. Metabolic components of asperosaponin VI after exposing to zebrafish for 24 h were identified by high performance liquid chromatography--electrospray mass spectrometry (HPLC-ESI-MS), the separation was performed with a Zorbax C18 column using a binary gradient elution of 0.05% formic acetonitrile--0.05% formic acid water. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures were identified by attentive study on the deglycosylation metabolites and one hydroxylation metabolite of asperosaponin VI. The results were highly in consistent with metabolism of asperosaponin VI in rat. It can be concluded that zebrafish model can wonderfully imitate current metabolic model with advantages of small amount of lower cost, far less amount compound, higher efficiency and more simple, and can reflect integrated metabolism results of in vivo method. Zebrafish metabolic model may become a novel organism model for quick predication on metabolism of even mircoamount compound, which can enrich the available models greatly.
Animals ; Chromatography, High Pressure Liquid ; Dipsacaceae ; chemistry ; Female ; Male ; Models, Animal ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Saponins ; isolation & purification ; metabolism ; Spectrometry, Mass, Electrospray Ionization ; Zebrafish ; metabolism