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Background: Osteoporosis is a prevalent bone metabolism disease characterized by a reduction in bone density, leading to several complications that significantly affect patients’ quality of life. The Achyranthes bidentata–Dipsacus asper (AB–DA) herb pair is commonly used in Traditional Chinese Medicine (TCM) to treat osteoporosis. This study aimed to investigate the therapeutic compounds and potential mechanisms of AB–DA using network pharmacology, molecular docking, molecular dynamics simulation, and experimental verification. Methods: Identified compounds of AB–DA were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Traditional Chinese Medicine Information Database (TCM-ID), TCM@ Taiwan Database, BATMAN-TCM, and relevant literature. The main bioactive ingredients were screened based on the criteria of “OB (oral bioavailability) ≥ 30, DL (drug-likeness) ≥ 0.18.” Potential targets were predicted using the PharmMapper and SwissTargetPrediction websites, while disease (osteoporosis)-related targets were obtained from the GeneCards, DisGeNET, and OMIM databases. The PPI network and KEGG/GO enrichment analysis were utilized for core targets and pathway screening in the STRING and Metascape databases, respectively. A drug–compound–target–pathway–disease network was constructed using Cytoscape software to display core regulatory mechanisms. Molecular docking and dynamics simulation techniques explored the binding reliability and stability between core compounds and targets. In vitro and in vivo validation experiments were utilized to explore the anti-osteoporosis efficiency and mechanism of sitogluside. Results: A total of 31 compounds with 83 potential targets for AB–DA against osteoporosis were obtained. The PPI analysis revealed several hub targets, including AKT1, CASP3, EGFR, IGF1, MAPK1, MAPK8, and MAPK14. GO/KEGG analysis indicated that the MAPK cascade (ERK/JNK/p38) is the main pathway involved in treating osteoporosis. The D–C–T–P–T network demonstrated therapeutic compounds that mainly consisted of iridoids, steroids, and flavonoids, such as sitogluside, loganic acid, and β-ecdysterone. Molecular docking and dynamics simulation analyses confirmed strong binding affinity and stability between core compounds and targets. Additionally, the validation experiments showed preliminary evidence of antiosteoporosis effects. Conclusion: This study identified iridoids, steroids, and flavonoids as the main therapeutic compounds of AB–DA in treating osteoporosis. The underlying mechanisms may involve targeting core MAPK cascade (ERK/JNK/p38) targets, such as MAPK1, MAPK8, and MAPK14. In vivo experiments preliminarily validated the anti-osteoporosis effect of sitogluside. Further in-depth experimental studies are required to validate the therapeutic value of AB–DA for treating osteoporosis in clinical practice. [ABSTRACT FROM AUTHOR]

Sweating is one of the most important primary processing methods of Chinese medicinal materials. Dipsacus asper is a typical representative of sweating treatment that is recommended by the Chinese Pharmacopoeia. The color change of the fracture surface of the root is the prominent feature of sweating treatment. However, few studies have focused on the mechanism of color change during sweating treatment. In this study, widely targeted metabolomics and ITS high-throughput sequencing technologies were applied to detect metabolites and microbial structure and diversity in the root of D. asper during sweating treatment. A total of 667 metabolites, including 36 downregulated and 78 upregulated metabolites, were identified in D. asper following sweating treatment. The significantly differential metabolites were divided into 12 classes, including terpenoids and phenolic acids. Moreover, all the differential terpenoids were upregulated and 20 phenolic acids showed a significant change after sweating treatment. In addition, microbial community diversity and richness increased following sweating treatment. The composition of microbial communities revealed that the relative abundances of Ascomycota and Basidiomycota significantly changed after sweating treatment. Correlation analysis revealed that Ascomycota (Fusarium sp., Macrophomina sp., Ilyonectria sp., Memnoniella sp., Penicillium sp., Cyphellophora sp., Neocosmospora sp., unclassified_f_Nectriaceae, and unclassified_o_Saccharomycetales) and Basidiomycota (Armillaria sp.) were associated with the content of terpenoids (6-deoxycatalpol and laciniatoside III) and phenolic acids (3-(4-hydroxyphenyl)-propionic acid, ethyl caffeate, 4-O-glucosyl-4-hydroxybenzoic acid, 2-acetyl-3-hydroxyphenyl-1-O-glucoside, 4-O-glucosyl-3,4-dihydroxybenzyl alcohol, 3-O-feruloylquinic acid, 3,4-O-dicaffeoylquinic acid methyl ester, O-anisic acid, and coniferyl alcohol). We speculate that the Ascomycota and Basidiomycota affect the content of terpenoids and phenolic acids, resulting in color change during sweating treatment in D. asper. This study provides a foundation for analyzing the mechanism involved in the processing of Chinese medicinal materials. [ABSTRACT FROM AUTHOR]

Recurrent spontaneous abortion (RSA) is a common pregnancy disease mostly occurring in the first trimester of pregnancy, and unexplained RSA (URSA) occurs in approximately 40% of RSA patients even after careful examination. Here, we assessed the effects of modified Bushen Huoxue Prescription combined with active immunotherapy on immunomodulatory functions and pregnancy outcomes of URSA patients. Ninety-six eligible URSA patients were randomly divided into control and experimental groups (n=48). Active immunotherapy was performed for control group, while experimental group were treated with modified Bushen Huoxue Prescription combined with active immunotherapy. Counts of peripheral blood helper T lymphocytes 17 (Th17) and regulatory T lymphocytes (Treg), levels of inflammatory factors interleukin-4 (IL-4), IL-10 and IL-17, and mRNA expression levels of retinoic acid-related orphan receptor γt (RORγt) and forkhead box P3 (FoxP3) were compared. Compared with before treatment, ratios of Th17/CD4+ cells and Th17/Treg, IL-17 level and RORγt mRNA level decreased, whereas ratio of Treg/CD4+ cells, IL-4 and IL-10 levels and FoxP3 mRNA level increased in both groups after treatment (P<0.05). After treatment, ratios of Th17/CD4+ cells and Th17/Treg, IL-17 level and RORγt mRNA level were lower in experimental group than those in control group, while ratio of Treg/CD4+ cells, IL-4, IL-10 levels and FoxP3 mRNA level were higher in experimental group (P<0.05). Modified Bushen Huoxue Prescription combined with active immunotherapy worked well for URSA to improve pregnancy outcomes and lower abortion rate. [ABSTRACT FROM AUTHOR]

This paper aims to study the spectrum-effect relationship between the fingerprints before and after salt processing of Dipsacus asper and the efficacy of warming and tonifying kidney Yang and find the main active components against kidney Yang deficiency before and after salt processing of D. asper, so as to provide the basis for clarifying the effect of salt processing on kidney Yang deficiency. The HPLC fingerprint before and after salt processing of D. asper was established by the HPLC-DAD. 15 common peaks were obtained, and 11 components were identified. The content changes of various components in rat serum were detected, and the difference in efficacy before and after salt processing was compared. The results of pharmacological experiments showed that salt processing of D. asper could enhance the kidney index. At the same dose, there was a significant difference between the raw D. asper and D. asper after salt processing groups. Compared with the model group, the contents of ACTH, cAMP, CORT, E&#95;2, GH, Na~+-K~+-ATPase, T, and T4 in the serum of rats in the administration group increased to a certain extent, and the contents of cGMP and TNF-α decreased to a certain extent. Among them, there were significant differences in the above indexes in the serum of rats in the high-dose group of raw D. asper, middle-dose group of D. asper after salt processing, high-dose group of D. asper after salt processing, and the positive drug group. The overall results showed that D. asper after salt processing was more effective than raw D. asper in preventing kidney yang deficiency. The efficacy of D. asper was evaluated by grey correlation analysis, entropy method, and Pearson correlation analysis, and the components of D. asper after salt processing against kidney yang deficiency were screened out. According to the results of correlation degree ranking, the components with increased ranking before and after salt processing of D. asper were loganin, chlorogenic acid, dipsacoside A, asperosaponin Ⅵ, caffeic acid, and isochlorogenic acid B. It was preliminarily speculated that these compounds may be the potential pharmacodynamic components for the treatment of kidney yang deficiency before and after salt processing of D. asper. The changing components before and after the salt processing of D. asper were determined, which proved that D. asper after salt processing was superior to D. asper in the treatment of kidney yang deficiency. The spectrum-effect relationship between the efficacy of D. asper before and after salt processing and the treatment of kidney yang deficiency was established, which laid a foundation for the subsequent study on the pharmacodynamic components and molecular mechanism of salt processing of D. asper.

• Three new and four known polymer iridoid glycosides were isolated from Dipsacus asper. • All of them were found to be inactive in anti-inflammatory activity. • Only D. asper has been reported isolated tetrairidoid glucosides thus far. Two new iridoid tetramers with four glucosides, dipsanosides H (1) and I (2), as well as one new bis-iridoid glucosides dipsanosides J (3), along with four known polymer iridoid glycosides dipsanosides A (4) and B (5), pterocephaline (6), triplostoside A (7) were isolated from Dipsacus asper. Their structures were determined by analysis of 1D and 2D NMR data. All of them were found to be inactive in anti-inflammatory activity. [ABSTRACT FROM AUTHOR]

The effects of a polysaccharide (DAP) from the roots of Dipsacus asper on renal complication in streptozotocin (STZ)-induced diabetic rats were investigated in the present study. Administration of DAP (100 and 300 mg/kg) for 4 weeks significantly decreased the fasting blood glucose (FBG) and glycosylated hemoglobin (HbA1c) levels, but increase the body weight of STZ-induced diabetic rats. Increased kidney weight index, serum creatinine (Scr), blood urea nitrogen (BUN), urine protein, and urinary albumin excretion (UAE) levels along with decreased creatinine clearance (Ccr) were nearly reversed to the normal levels by DAP. Moreover, DAP was able to normalize hyperlipidemia, improve oxidative stress, and down-regulate the formation of advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE). In conclusion, our work demonstrates the preventive effect of DAP was achieved via antihyperglycemic, hypolipidemic, and antioxidant activity along with inhibition of AGE accumulation and RAGE expression in STZ-treated diabetic rats. [ABSTRACT FROM AUTHOR]

Iridoid-monoterpenoid indole alkaloid hybrids (IMIAHs) represent a rare class of natural products reported from only several plants of Rubiaceae and Dipsacaceae families, while their structural assignments remain a very challenging work due to complexity and flexibility. In the current study, a new IMIAH (1) was isolated from the roots of Dipsacus asper and its structure with absolute configuration was unambiguously established by a combination of spectroscopic analyses, chemical degradation and ECD calculation. A new oleanane-type triterpenoid saponin (2) and 15 known co-metabolites were also obtained and structurally characterized. Our biological evaluations showed that compound 2 exhibited moderate inhibition against acetylcholine esterase (AChE) with an IC 50 value of 15.8 ± 0.56 μM, and compound 15 displayed potent cytotoxicity selectively against human A549 and H157 lung cancer cells with IC 50 values of 6.94 ± 0.24 and 9.06 ± 0.12 μM, respectively. Unlabelled Image [ABSTRACT FROM AUTHOR]

Phytochemical investigation of the ethyl acetate soluble part, generated from the ethanol extract of the roots of Dipsacus asper , led to the separation and identification of three undescribed triterpenoids including one arborinane type, one ursane type and one oleanane type, two unreported oleanane type triterpenoid arabinoglycosides, and 18 known analogues. Structures of these compounds were determined by comprehensive spectroscopic analyses, with the absolute configurations of 25-acetoxy-28-dehydroxyrubiarbonone E and 2 α ,3 β -dihydroxy-23-norurs-4(24),11,13(18)-trien-28-oic acid being established by evaluation of their experimental and calculated ECD spectra. 25-Acetoxy-28-dehydroxyrubiarbonone E features an oxygenated C-25 that is the first case among arborinane type triterpenoids, while 2 α ,3 β ,24-trihydroxy-23-norurs-12-en-28-oic acid incorporates a sp3 C-24 that is a rare structural feature of 23-norursane type triterpenoids. Of these isolates, 2′,4′- O -diacetyl-3- O - α - l -arabinopyranosyl-23-hydroxyolea-12-en-28-oic acid and hederagonic acid exhibited moderate antibacterial activity against Staphylococcus aureus with IC 50 values of 12.3 and 10.3 μ M, respectively, while those with either a feruloyloxy group or an arabinosyl moiety at C-3 displayed potent cytotoxic activities against four tumor cell lines A549, H157, HepG2 and MCF-7. Twenty-three compounds including five undescribed ones were isolated from the roots of Dipsacus asper and their cytotoxic evaluation with SAR discussion was presented. Image 1 • Five undescribed compounds were isolated from the roots of Dipsacus asper. • TDDFT calculations of ECD spectra were used to determine the absolute configurations. • Some compounds exhibited moderate antibacterial activities. • Some isolates displayed potent cytotoxic activities with IC 50 values below 10 μ M. • The structure-activity relationship (SAR) for cytotoxic evaluation was well discussed. [ABSTRACT FROM AUTHOR]

Background: Osteoporosis is a prevalent bone metabolism disease characterized by a reduction in bone density, leading to several complications that significantly affect patients' quality of life. The Achyranthes bidentata-Dipsacus asper (AB-DA) herb pair is commonly used in Traditional Chinese Medicine (TCM) to treat osteoporosis. This study aimed to investigate the therapeutic compounds and potential mechanisms of AB-DA using network pharmacology, molecular docking, molecular dynamics simulation, and experimental verification. Methods: Identified compounds of AB-DA were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Traditional Chinese Medicine Information Database (TCM-ID), TCM@Taiwan Database, BATMAN-TCM, and relevant literature. The main bioactive ingredients were screened based on the criteria of "OB (oral bioavailability) ≥ 30, DL (drug-likeness) ≥ 0.18." Potential targets were predicted using the PharmMapper and SwissTargetPrediction websites, while disease (osteoporosis)-related targets were obtained from the GeneCards, DisGeNET, and OMIM databases. The PPI network and KEGG/GO enrichment analysis were utilized for core targets and pathway screening in the STRING and Metascape databases, respectively. A drug-compound-target-pathway-disease network was constructed using Cytoscape software to display core regulatory mechanisms. Molecular docking and dynamics simulation techniques explored the binding reliability and stability between core compounds and targets. In vitro and in vivo validation experiments were utilized to explore the anti-osteoporosis efficiency and mechanism of sitogluside. Results: A total of 31 compounds with 83 potential targets for AB-DA against osteoporosis were obtained. The PPI analysis revealed several hub targets, including AKT1, CASP3, EGFR, IGF1, MAPK1, MAPK8, and MAPK14. GO/KEGG analysis indicated that the MAPK cascade (ERK/JNK/p38) is the main pathway involved in treating osteoporosis. The D-C-T-P-T network demonstrated therapeutic compounds that mainly consisted of iridoids, steroids, and flavonoids, such as sitogluside, loganic acid, and β-ecdysterone. Molecular docking and dynamics simulation analyses confirmed strong binding affinity and stability between core compounds and targets. Additionally, the validation experiments showed preliminary evidence of antiosteoporosis effects. Conclusion: This study identified iridoids, steroids, and flavonoids as the main therapeutic compounds of AB-DA in treating osteoporosis. The underlying mechanisms may involve targeting core MAPK cascade (ERK/JNK/p38) targets, such as MAPK1, MAPK8, and MAPK14. In vivo experiments preliminarily validated the anti-osteoporosis effect of sitogluside. Further in-depth experimental studies are required to validate the therapeutic value of AB-DA for treating osteoporosis in clinical practice., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Li, Li, Guo, Tan, Xiang and Ouyang.)

Sweating is one of the most important primary processing methods of Chinese medicinal materials. Dipsacus asper is a typical representative of sweating treatment that is recommended by the Chinese Pharmacopoeia. The color change of the fracture surface of the root is the prominent feature of sweating treatment. However, few studies have focused on the mechanism of color change during sweating treatment. In this study, widely targeted metabolomics and ITS high-throughput sequencing technologies were applied to detect metabolites and microbial structure and diversity in the root of D. asper during sweating treatment. A total of 667 metabolites, including 36 downregulated and 78 upregulated metabolites, were identified in D. asper following sweating treatment. The significantly differential metabolites were divided into 12 classes, including terpenoids and phenolic acids. Moreover, all the differential terpenoids were upregulated and 20 phenolic acids showed a significant change after sweating treatment. In addition, microbial community diversity and richness increased following sweating treatment. The composition of microbial communities revealed that the relative abundances of Ascomycota and Basidiomycota significantly changed after sweating treatment. Correlation analysis revealed that Ascomycota ( Fusarium sp., Macrophomina sp., Ilyonectria sp., Memnoniella sp., Penicillium sp., Cyphellophora sp., Neocosmospora sp., unclassified&#95;f&#95;Nectriaceae, and unclassified&#95;o&#95;Saccharomycetales) and Basidiomycota ( Armillaria sp.) were associated with the content of terpenoids (6-deoxycatalpol and laciniatoside III) and phenolic acids (3-(4-hydroxyphenyl)-propionic acid, ethyl caffeate, 4-O-glucosyl-4-hydroxybenzoic acid, 2-acetyl-3-hydroxyphenyl-1-O-glucoside, 4-O-glucosyl-3,4-dihydroxybenzyl alcohol, 3-O-feruloylquinic acid, 3,4-O-dicaffeoylquinic acid methyl ester, O-anisic acid, and coniferyl alcohol). We speculate that the Ascomycota and Basidiomycota affect the content of terpenoids and phenolic acids, resulting in color change during sweating treatment in D. asper. This study provides a foundation for analyzing the mechanism involved in the processing of Chinese medicinal materials., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 He, Xu, Zhou, Yang, Yang, Xiao, Zhang, Li and Zhou.)

Radix Dipsaci (RD), the dried root of Dipsacus asper , is used in traditional Chinese medicine as a remedy for bone fractures, traumatic hematoma, threatened abortion, and uterine bleeding. A novel ultra high-performance liquid chromatography coupled with quadrupole-time-of-flight tanderm mass spectrometry (UHPLC-Q-TOF/MS) approach was performed to rapidly characterize the chemical constituents of RD. Consequently, 21 compounds, including 12 iridoid glycosides (IGs), 4 furofuran lignans (FLs), and 5 phenolic acids (PAs) were discovered and identified from RD. Among these compounds, 3 IGs were previously unreported. Furthermore, a rapid and reliable UHPLC-DAD-based method was developed. The linearity (R 2  > 0.999), repeatability (RSDs < 3.0%), intra-day and inter-day precision (RSDs < 1.6%), recovery (98.9%–102.5%), limits of detection (0.2–2.75 ng), and limits of quantification (0.75–8.5 ng) of the method was validated. The method was successfully applied for the simultaneous determination of 11 compounds in 20 batches of RD collected from various geographical regions in China. Different RD samples exhibited significantly varied contents of 11 analytes, among which IGs and PAs are abundant compounds that could be used as suitable quality markers for RD. The present study provided a useful approach for comprehensively evaluating the chemical composition and quality of RD. [ABSTRACT FROM AUTHOR]

The in vivo antifungal activity of crude extracts of Dipsacus asper roots was evaluated against the phytopathogenic fungi Botrytis cinerea , Colletotrichum coccodes , Blumeria graminis f. sp. hordei , Magnaporthe grisea , Phytophthora infestans , Puccinia recondita and Rhizoctonia solani using a whole-plant assay method. Ethyl acetate and acetone extracts, at 1000 μg/mL, suppressed the development of tomato gray mold (TGM) and tomato late blight (TLB) by 90%. Through bioassay-guided isolation, five antifungal substances were isolated from the D. asper roots and identified as β -sitosterol ( 1 ), campesterol ( 2 ), stigmasterol ( 3 ), cauloside A ( 4 ) and a novel dipsacus saponin, named colchiside (3- O - β - d -xylopyranosyl-23- O - β - d -glucopyranosyl-28- O - β - d -(6- O -acetyl)-glucopyranosyl hederagenin) ( 5 ). Of those, cauloside A ( 4 ) displayed the greatest antifungal efficacy against rice blast, TGM and TLB. Colchiside ( 5 ) moderately suppressed the development of TLB, but exhibited little effect against the other diseases. The synergistic effects of the isolated compounds against TLB were also assessed. Synergistic and additive interactions were observed between several of the sterol compounds. This study indicated that the crude extracts of, and bioactive substances from, the roots of D. asper suppress TGM and TLB. In addition, cauloside A ( 4 ) and colchiside ( 5 ) could be used as antifungal lead compounds. [ABSTRACT FROM AUTHOR]

Wine-processing, which is sauteing with rice wine, will change the inclination and direction of herbs’ actions. After being wine-processed, the effects of nourishing liver and kidney of Dipsacus asper will be strengthened. However, the underlying mechanism remains elusive. The following study is to establish and validate an UHPLC–MS/MS approach to determine six bioactive constituents in tissue samples, including loganin, loganic acid, chlorogenic acid, 3,5-dicaffeoylquinic acid, 4-caffeoylquinic acid and asperosaponin VI and apply the approach to a comparative tissue distribution study of raw and wine-processed Dipsacus asper in rats. A Shimadzu UHPLC system coupled with triple quadrupole mass spectrometer was employed for analysis of the six analytes using multiple reaction monitoring (MRM) mode. A one-step protein precipitation by methanol was employed to extract the six analytes from tissues. Chloramphenicol and glycyrrhetinic acid were selected as internal standards. The proposed approach was fully validated in terms of linearity, sensitivity, precision, repeatability as well as recovery. Our results revealed that all of the calibration curves displayed good linear regression (r 2 > 0.9991). Intra- and inter-assay variability for all analytes ranged from −4.62 to 4.93% and from −4.98 to 4.92%, respectively. The recovery rates for each analytes were determined to be 88.3–100.1%. All the samples showed satisfactory precision and accuracy after various stability tests, including storage at 25 °C for 4 h, −80 °C for 30 days, three-freeze-thaw cycles, and 4 °C for 24 h. Tissue pharmacokinetic parameters including AUC 0-t , t 1/2 , T max and C max were calculated. Collectively, the parameters of C max and AUC 0-t of the six analytes in wine-processed group were remarkably elevated ( p < 0.05) in the rat liver and kidney as compared with those of the raw group. But in the rat heart and spleen, the C max and AUC 0-t of asperosaponin VI was decreased as compared with those of the raw group. The accumulation of bioactive constituents in liver and kidney tissues after wine-processing will contribute to the enhancement of liver and kidney nourishing effects. [ABSTRACT FROM AUTHOR]

A rapid and sensitive ultra-high performance liquid chromatography with tandem mass spectrometry approach was established for the simultaneous determination of 4-caffeoylquinic acid, loganic acid, chlorogenic acid, loganin, 3,5-dicaffeoylquinic acid, dipsacoside B, asperosaponin VI, and sweroside in raw and wine-processed Dipsacus asper. Chloramphenicol and glycyrrhetinic acid were employed as internal standards. The proposed approach was fully validated in terms of linearity, sensitivity, precision, repeatability as well as recovery. Intra- and interassay variability for all analytes were 2.8-4.9 and 1.7-4.8%, respectively. The standard addition method determined recovery rates for each analytes (96.8-104.6%). In addition, the developed approach was applied to 20 batches of raw and wine-processed samples of Dipsacus asper. Principle component analysis and partial least squares-discriminate analysis revealed a clear separation between the raw group and wine-processed group. After wine-processing, the contents of loganic acid, chlorogenic acid, dipsacoside B, and asperosaponin VI were upregulated, while the contents of 3,5-dicaffeoylquinic acid, 4-caffeoylquinic acid, loganin, and sweroside were downregulated. Our results demonstrated that ultra-high performance liquid chromatography with tandem mass spectrometry quantification combined with chemometrics is a viable method for quality evaluation of the raw Dipsacus asper and its wine-processed products. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the larvicidal activity of the essential oil of Dipsacus asper Wallich ex Candolle (Dipsacaceae) roots against the larvae of Aedes aegypti L. and Culex pipiens pallens Coquillett. Methods: Essential oil was extracted from D. asper roots by hydrodistillation and analyzed for its composition by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The oil was evaluated for larvicidal activity, using World Health Organization (WHO) procedures, against the fourth larvae of A. aegypti and C. pipiens pallens within 24 h, and activity was recorded for various concentrations of the oil ranging from 12.5 - 200.0 µg/mL. Results: A total of 34 components of the essential oil of D. asper were identified. The major compounds of the essential oil were caryophyllene oxide (13.29 %), caryophyllene (9.14 %), cubebene (7.87 %), ß-gurjunene (6.43 %), carvone (5.38 %), 1,8-cineole (5.29 %), and calamenene (5.05 %). The oil exhibited larvicidal activity against A. aegypti and C. pipiens pallens at median lethal concentrations (LC50) of 56.29 µg/mL and 47.49 µg/mL, respectively. Conclusion: The essential oil of D. asper roots has potentials for use in the control of A. aegypti and C. pipiens pallens and may be useful in the search for new, safer and more effective natural larvicides. [ABSTRACT FROM AUTHOR]

The purpose of this study is to establish and validate an UPLC–MS/MS approach to determine 4-caffeoylquinic acid, chlorogenic acid, 3,5-dicaffeoylquinic acid, loganic acid, loganin, sweroside, dipsacoside B and asperosaponin VI from extracts of crude and wine-processed Dipsacus asper in biological samples and apply the approach to a comparative pharmacokinetic study. A Waters BEH C 18 UPLC column was employed with acetonitrile/0.2% formic acid-water as mobile phases. The mass analysis was carried out in a triple quadrupole mass spectrometer using multiple reaction monitoring (MRM) with negative scan mode. A one-step protein precipitation by acetonitrile was performed to extract the eight analytes from plasma. Our results revealed that all of the calibration curves displayed good linear regression (r 2 > 0.9990). The lower limits of quantification (LLOQ) were determined as 10.0, 9.6, 8.9, 9.1, 9.2, 9.8, 10.1 and 9.8 ng/mL. The intra-day and inter-day precisions (RSD) of the eight compounds at high, medium and low levels were less than 4.94% and the bias of the accuracies ranged from −3.89% to 3.95%.The extraction recoveries of the eight compounds were from 90.4% to 100.2% and the matrix effects ranged from 89.3% to 100.1%. The stabilities of these compounds were investigated by analyzing six replicates of QC samples at three different concentrations following storage at 25 °C for 4 h, −80 °C for 30 days, three-freeze-thaw cycles, and 4 °C for 24 h. All the samples showed satisfactory precision and accuracy after various stability tests. Pharmacokinetic parameters were estimated using a non-compartment model. Compared with the crude group, the parameters of C max and AUC 0-t of 4-caffeoylquinic acid, loganic acid, loganin and asperosaponin VI increased remarkably ( p < 0.05 ) after oral administration of the aqueous extract of wine-processed Dipsacus asper , indicating that wine-processing could enhance bioavailability of 4-caffeoylquinic acid, loganic acid, loganin and asperosaponin VI. [ABSTRACT FROM AUTHOR]

A new iridoid glycoside, named loganic acid ethyl ester (1), together with five known compounds: chlorogenic acid (2), caffeic acid (3), loganin (4), cantleyoside (5) and syringaresinol-4′,4′′-O-bis-β-D-glucoside (6) were isolated from the roots of Dipsacus asper. The structure of compound 1 was elucidated on the basis of detailed spectroscopic analyses. Lignan is isolated from Dipsacaceae species for the first time. Compounds 1, 4 and 5 had moderate neuroprotective effects against the Aβ25–35 induced cell death in PC12 cells.

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.

Amethanolic extract of Dipsacus asper, having anti-diabetic activity, was examined as a possible aldose reductase (ALR2) inhibitor, a key enzyme involved in diabetic complications. Bioactivity guided fractionation led to the isolation of ten compounds, ursolic acid (1), oleanolic acid-3- O-α-L-arabinopyranoside (2), daucosterol (3), hederagenin-3- O-α-L-arabinopyranoside (4), sweroside(5), caffeic acid (6), esculetin (7), protocatechualdehyde (8), loganin (9), and vanilic acid (10) from the ethyl acetate fraction of D. asper methanol extract. Among them, compounds 4, 6, 7, and 8 exhibited inhibitory effects on aldose reductase, with IC values of 23.70, 16.71, 34.36, and 21.81 μM, respectively. This is the first report on the isolation of these compounds from D. asper, and the ALR2 inhibitory activity of hederagenin-3- O-α-L-arabinopyranoside. These results suggest the successful use of the extract of D. asper for ameliorating diabetic complications. [ABSTRACT FROM AUTHOR]

Abstract: Three new triterpene saponins, named asperosaponin A–C (1–3), together with seven known triterpene saponins (4–10) were isolated from the roots of Dipsacus asper. The structures of compounds 1–3 were elucidated on the basis of detailed spectroscopic analysis and chemical methods. Compounds 1–3, 6–10 had significantly protective effects in PC12 cells against the Aβ25-35 induced cytotoxicity. All the compounds isolated showed no cytotoxic activity against three human cancer cell lines, A-549, Bel-7402 and BGC-823. [Copyright &y& Elsevier]

A new iridoid glycoside, named loganic acid ethyl ester (1), together with five known compounds: chlorogenic acid (2), caffeic acid (3), loganin (4), cantleyoside (5) and syringaresinol-4',4''-O-bis-β-D-glucoside (6) were isolated from the roots of Dipsacus asper. The structure of compound 1 was elucidated on the basis of detailed spectroscopic analyses. Lignan is isolated from Dipsacaceae species for the first time. Compounds 1, 4 and 5 had moderate neuroprotective effects against the A#x03B2;25-35 induced cell death in PC12 cells. [ABSTRACT FROM AUTHOR]

Two new triterpenoid saponins 1 and 2, along with six known saponins 3-8, were isolated from the roots of Dipsacus asper. The structures of new compounds were established as 3-O-β-d-glucopyranosyl-(1 → 4)-[α-l-rhamnopyranosyl-(1 → 3)]-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 2)-α-l-arabinopyranosyl-hederagenin-28-O-β-d-glucopyranoside (dipsacus saponin J, 1) and 3-O-α-l-arabinopyranosyl-hederagenin-28-O-β-d-glucopyranosyl-(1 → 6)-β-d-glucopyranosyl-(1 → 6)-β-d-glucopyranoside (dipsacus saponin K, 2). The structures were determined by extensive analysis of their spectroscopic data. Compounds 6 and 7 could significantly stimulate UMR106 cell proliferation and increase alkaline phosphatase activities in UMR106 cell at the concentration of 4 μM. [ABSTRACT FROM AUTHOR]

Objective To compare the regulation difference of gavage administration of crude and wine-processed Dipsacus on OPG/RANK/RANKL axis in osteoporosis rat. Methods Thirty SPF grade SD rats purchased from the Shanghai Jiesijie Laboratory Animal Co., Ltd. were established the osteoporotic model by the castration method and randomly divided into the normal group, sham operation group,model group, estradiol group, crude Dipsacus group and wine-processed Dipsacus group, 5 cases in each group. After 12 weeks of gavage administration, the rats were anesthetized and aortic blood was collected. The basic biochemical indicators of serum Ca, P, alkaline phosphatase and osteocalcin were analyzed by adopting the automatic biochemical analyzer. Meanwhile, the 3 bone metabolic indicators of serum OPG, RANK and RANKL levels were determined by using the ELISA method. Results Compared with the model group, the levels of alkaline phosphatase, osteocalcin and RANKL in the estradiol group, crude Dipsacus group and wine-processed Dipsacus group were significantly reduced, while the levels of OPG and RANK were significantly increased, the Ca level in the wine-processed Dipsacus group was significantly reduced and the P level was increased,the differences were statistically significant (P<0.05); compared with the estradiol group, the levels of alkaline phosphatase and osteocalcin in the crude Dipsacus group were significantly increased, whereas the Ca level in the wine-processed Dipsacus group was significantly reduced and the levels of P and osteocalcin were significantly increased, the levels of OPG and RANK in the crude and wine-processed Dipsacus groups were significantly reduced, while the RANKL level was significantly elevated,the differences were statistically significant (P<0.05); compared with the crude Dipsacus group, the levels of Ca,alkaline phosphatase,osteocalcin and RANKL levels in the wine-processed Dipsacus group were significantly reduced, while the levels of OPG and RANK were significantly increased with statistical differences (P<0.05). Conclusion The curative effect of Dipsacus in treating osteoporosis rat is related with its regulating the OPG/RANK/RANKL axis. The regulation ability of wine-processed Dipsacus is stronger than that of crude Dipsacus. [ABSTRACT FROM AUTHOR]

Methanol extracts of the root of Dipsacus asper Wall (Dipsacaceae) were found to exhibit apoptosis-inducing activities in U937 (human monocyte-like histiocytic) cells. Investigation of the active n-BuOH fraction led to the isolation of akebia saponin D (ASD). Structure was established by spectroscopic methods. Treatment of U937 cells with ASD induced apoptosis in a dose dependent manner. ASD exerted strong cytotoxicity against human and murine leukemia cells. It is significantly increased the subG1 cell population and expression of p53 and Bax gene. And also ASD enhanced NO production from RAW264.7 macrophage cells. Taken together, these results strongly indicate that ASD may exert apoptosis-inducing activity via induction of apoptosis through activation chiefly via the nitric oxide and apoptosis-related p53 and Bax gene expression. These data provide scientific evidence that Dipsacus asper Wall can be useful as a chemopreventive agent. [ABSTRACT FROM AUTHOR]

Cytotoxic activity of seven hederagenin saponins isolated from the root of Dipsacus asper were investigated in vitro against L1210, HL-60, and SK-OV-3 tumor cell lines by the MTT method. 3- O-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin ( 2), 3- O-β-D-xylopyranosyl-(1→3)-α-L-Rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin ( 6) and 3- O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin ( 7) exhibited the potent cytotoxicity against the three tumor cell lines with IC50 values ranging from 4.7 to 8.7 μg/mL, with the exception of compound 7, which exhibited weak cytotoxic activity against SK-OV-3 (IC50 22.5 μg/mL). Other compounds did not exhibit any cytotoxic activity (IC50>30 μg/mL). [ABSTRACT FROM AUTHOR]

Excess aluminum (Al) exposure impairs neurocognitive function in humans and animals. Epidemiologic studies have shown a potential link between chronic Al exposure and Alzheimer''s disease. In the present study, we sought to evaluate the protective effects of the herbal medicine Dipsacus asper extract against the cognitive impairment and overexpression of hippocampal β-amyloid protein (Aβ) induced by chronic Al exposure in rats. Vitamin E (VE) was used as a positive control. Following exposure to 0.3% aluminum chloride (AlCl3) solution for 90 days in their drinking water, animals displayed a striking decrease (>80%) in step-through latency in the passive avoidance task and a significant increase (123%) in the number of Aβ immunoreactive cells in the hippocampus compared to controls. Al-exposed animals were then randomly assigned to receive vehicle, Dipsacus asper extract (4 g/kg), or VE (40 mg/kg) treatment up to 5 months. Both Dipsacus asper extract and VE significantly ameliorated animal''s performance impairment in the passive avoidance task and suppressed the overexpression of hippocampal Aβ immunoreactivity. The effects of Dipsacus asper extract, but not VE, increased with time of treatment. The present results suggest that Dipsacus asper extract may possess therapeutic effects against Alzheimer''s disease. [Copyright &y& Elsevier]

Four triterpenoids were isolated from the roots of Dipsacus asper. On the basis of chemical and spectral evidence, the structures of these compounds have been elucidated to be hederagenin ( 1), hederagenin 3- O-α-L-arabinoside ( 2). 3- O-α-L-arabinopyranosyl hederagenin 28- O-β-D-glucopyranosyl ester ( 3) and hederagenin 28- O-β-D-glucopyranosyl (1→6)-β-D-glucopyranosyl ester ( 4). The new glycoside, 4, was named dipsacus saponin A. [ABSTRACT FROM AUTHOR]

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.

Anticomplementary activity of hederagenin and related saponins isolated from Dipsacus asper was investigated in vitro. HN saponin F ( 3) was most potent with IC50 value of 3.7×10−5 M followed by 3- O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-β-L-arabinopyranosyl hederagenin 28- O-β-D-glucopyranosyl-(1→6)-β-D-glucopyrano side ( 8), 3- O-β-L-arabinopyranosyl hederagenin 28- O-β-D-glucopyranosyl-(1→6)-β-D-glucopyrano side ( 5) dipsacus saponin A ( 4), and hederagenin ( 1) on the classical pathway (CP) of complement system, while the saponins 3–5, did not show the inhibition of hemolysis and rather increase the hemolysis on the alternative pathway (AP). However, all of C-3 monodesmosides [prosapogenin CP ( 2), dipsacus saponin B ( 6), and dipsacus saponin C ( 7)] evoked hemolysis directly on the erythrocytes. [ABSTRACT FROM AUTHOR]