In order to study the distribution and dynamics growth of wild Dipsacus asper resources in the Wulong district of Chongqing, 9 sample plots were selected for 12 consecutive months in the natural distribution area of the D. asper in Wulong district by using the sample line + plot survey method to conduct a field survey. The results showed that D.asper was distributed in forest edge wasteland or shrub-grassland, and growbetter with loose yellow-brownsoil or red soil, and poor with lithologic soil or impounded surface water.The growth curve of the plant height from June to July and the ground fresh weight from July to August showed a turning point, it might consume large amounts of nutrients during its flowering period, resulting in the restriction of vegetative growth.The highest temperature in the distribution area of D.asperoides in Wulong district is less than 30 °C, the minimum temperature is about 0 °C, and the rainfall is 1 241-1 392 mm. Its growth environment is no severecold in winter, no heat in summer, and abundant rainfall.The main growth stage of D.asper is from July to October, and the range of root dry rate was 0.162 5-0.239 7 in Xiangkou, 0.154 9-0.223 6 in Baima Mountain, and 0.143 7-0.203 3 Xiannv Mountain. The vegetative growth and dry matter accumulation synchronized in the main growth stage, and the accumulation rate of dry matter was faster than that of vegetative growth. The correlation analysis between indicators and root fresh weight showed that the fresh weight of the aerial part and root fresh weight had the best correlation.
Dipsacaceae

The present study aimed to explore the mechanism of the sweating of Dipsacus asper on content changes of triterpene sa-ponins by detecting the total triterpene saponins and the index component asperosaponin Ⅵ in the crude and sweated D. asper, and analyzing the differentially expressed proteins by isobaric tags for relative and absolute quantification(iTRAQ) combined with LC-MS/MS. After sweating, the content of total triterpene saponins decreased manifestly, while that of asperosaponin Ⅵ increased significantly. As revealed by the iTRAQ-LC-MS/MS analysis, 140 proteins with significant differential expression were figured out, with 50 up-regulated and 90 down-regulated. GO analysis indicated a variety of hydrolases, oxido-reductases, and transferases in the differential proteins. The results of activity test on two differentially expressed oxido-reductases were consistent with those of the iTRAQ-LC-MS/MS analysis. As demonstrated by the analysis of enzymes related to the triterpene saponin biosynthesis pathway, two enzymes(from CYP450 and UGT families, respectively, which are involved in the structural modification of triterpene saponins) were significantly down-regulated after sweating. The findings suggested that sweating of D. asper presumedly regulated triterpene saponins by affecting the expression of downstream CYP450 s and UGTs in the biosynthesis pathway of triterpene saponins of D. asper.
Chromatography, Liquid ; Dipsacaceae ; Humans ; Saponins ; Sweating ; Tandem Mass Spectrometry ; Triterpenes

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

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

The author detected the genetic diversity and genetic relationship within and among eight medicinal species of Dipsacus by the approach of sequence-related amplified polymorphism (SRAP). The associated genetic parameters were calculated by POPGENE 1.31. The Genetic distance was calculated by TREECONW and the systematic diagrams of genetic relationship were clustered by UPG-MA. The results showed that, using 26 primers, a total of 558 bands were produced, of which 539 were polymorphic loci. There was a high level of genetic diversity among species (PPB = 96.59%, Na = 1.9659, Na = 1.3375, H = 0.2143, I = 0.3423). However, genetic diversity was lower within species, the average of genetic parameters was PPB = 6.97%, Na = 1.0697, Na = 1.0311, H = 0.0187, I = 0.0291. The Nei's genetic differentiation coefficient was 0.9126, indicated that most of the genetic variation existed among species. By clustering analysis, different individuals gathered in the same group and the classified result of SRAP marker between traditional modal characters was almost same. The results confirmed that SRAP marker can be used as one of the effective methods to reveal the genetic diversity and relationship among medicinal species of Dipsacus.
China ; Dipsacaceae ; classification ; genetics ; Gene Amplification ; Genetic Variation ; Plants, Medicinal ; classification ; genetics ; 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

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*

To study the chemical constituents of Dipsacus asper, chromatographic methods such as D101 macroporous resin, silica gel, octadecylsilyl (ODS) column chromatographic techniques and preparative HPLC were used, and five compounds were isolated from 70% (v/v) ethanol extract of the plant. By using spectroscopic techniques including 1H NMR, 13C NMR, 1H-1H COSY, HSQC, HMBC and TOF-MS, the compounds were identified as 3beta-hydroxy-24-nor-urs-4 (23), 12-dien-28-oic acid (1), ursolic acid (2), oleanolic acid (3), 3-O-alpha-L-rhamnosyl(1 --> 3)-beta-D-glucopyranosyl (1 --> 3)-alpha-L-rhamnosyl (1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranosyl ester (4), 3-O-[beta-D-xylopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 4)] [alpha-L-rhamnosyl(1 --> 3)]-beta-D-glucopyranosyl (1 --> 3)-alpha-L-rhamnosyl(1 --> 2)-alpha-L-arabinopyranosyl hederagenin (5), separately. Among them, 1 is a new compound, and 2 is isolated from this plant for the first time.
Dipsacaceae ; chemistry ; Molecular Structure ; Oleanolic Acid ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification

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

OBJECTIVETo develop a quantitative method for the determination of akebia saponin D in root of Dipacus asperoides and provide scientific basis for quality control of D. asperoides.

METHODAn IPLC analytical method was established using Kromasil ODS column with acetonitrile-water (30:70) as mobile phase and the detection wavelength was 212 nm.

RESULTThe akebia saponin D in root extract were well separated by this method. The linear range is between 0.582 5-9.32 microg, r = 0.9999. The average recovery and RSD of repeatability are 100.3%, 2.3%, respectively.

CONCLUSIONThis method is simple, accurate and reliable by evaluating the method validation data and can be used for the quality control of D. asperoides and its preparations.

China ; Chromatography, High Pressure Liquid ; Dipsacaceae ; chemistry ; Ecosystem ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Saponins ; analysis