Chinese Crude Drug Research Articles

Non-target metabolomic analysis reveals the therapeutic effect of Saposhnikovia divaricata decoction on collagen-induced arthritis rats

Ethnopharmacological relevanceSaposhnikovia divaricata (SD), a Chinese crude drug, has long been recognized for therapeutic effect to rheumatoid arthritis (RA). At present, the mechanisms of SD treatment in RA have not been fully understood especially on the perspective of metabolomics.Aim of the study: To study the pharmacodynamic effects of Saposhnikovia divaricata decoction on CIA rats, and explore the therapeutic mechanism by metabolomics methods. Materials and methodsWistar rats were randomly divided into normal group, CIA model group, dexamethasone group and SD decoction groups (10 g crude drug/kg, 5 g crude drug/kg and 2.5 g crude drug/kg of SDD). Body weight, arthritis scores, serum cytokine levels and histopathological parameters of rats were assessed. A metabolomics method based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOFMS) was established to collect the metabolic profiles of rats and explore the metabolic changes that occurred after SDD treatment. ResultsSDD showed its protective effect on the affected joints, especially in the middle dosage group of SDD. Eighteen and 13 potential biomarkers for the SDD treatment of CIA rats were identified in the plasma and urine, respectively. SDD could regulate the disturbed metabolic pathways including tryptophan metabolism, glycerophospholipid catabolism, primary bile acid biosynthesis and fatty acid metabolism. ConclusionsIn summary, SDD treatment could effectively alleviate symptoms of RA and regulate metabolic disorders in CIA rats.

UPLC/Q-TOF-MS/MS-based metabolomics revealed the lipid-lowering effect of Ilicis Rotundae Cortex on high-fat diet induced hyperlipidemia rats

Ethnopharmacological relevanceIlicis Rotundae Cortex (IRC), a Chinese crude drug, has been widely utilized in Guangdong and Guangxi provinces of China to treat or prevent cardiovascular diseases. Aim of studyThis investigation aims to study the lipid-lowering effect of IRC, as well as the regulating effect on the endogenous metabolites in hyperlipidemia rats. Materials and methodsHigh-fat diet induced hyperlipidemia rats were administrated with different doses of IRC extract (0.5, 1.0 and 2.0 g/kg/day) for 5 weeks. Simvastatin was used as the positive control. Body weight, serum lipid levels and histopathology of liver were evaluated. The metabolic profiles of plasma, urine and cecum content were analyzed using UPLC/Q-TOF-MS/MS-based metabolomics approach coupled with multivariate data analysis. ResultsThe levels of serum TC, TG, LDL-C, AST and ALT were significantly decreased and HDL-C level was increased in IRC treatment groups. The hepatic histomorphology was partially restored. 23, 26 and 15 metabolites in plasma, urine and cecum content were determined as the biological biomarkers, respectively. IRC extract could partially recover the disturbed metabolic pathways of bile acid metabolism, linoleic acid metabolism, arachidonic acid metabolism, taurine and hypotaurine metabolism, glyoxylate and dicarboxylate metabolism, glycerophospholipid metabolism, synthesis and degradation of ketone bodies, sphingolipid metabolism and riboflavin metabolism. ConclusionThis study demonstrated that IRC could effectively improve the serum lipids and partially restore the hepatic histomorphology. The underlying metabolic mechanism mainly included improving the metabolism of bile acids, glycerophospholipid, sphingolipid, fatty acid and amino acid. This is the first study on the lipid-lowering effect of IRC from the perspective of metabolomics.

Nor-kurarinone Characteristic of Chinese Sophora flavescens

The root of Sophora flavescens is used as a Chinese crude drug for treating various disorders. S. flavescens is grown in Japan, China and Korea. We previously used Principle Component Analysis (PCA) to demonstrate that 1H NMR spectra of root extracts of S. flavescens can be used to identify the country of origin of the sample (Japan or China). In the present study we identified a compound characteristic of Chinese S. flavescens using NMR metabolomics. Loading plot analysis of the PCA results and an NMR metabolomics study identified NMR signals from a compound identified as nor-kurarinone and characteristic of Chinese S. flavescens grown in Hangzhou. This compound was confirmed by HPLC analysis to be a distinctive marker for Chinese S. flavescens.

Steric Effect of Antioxidant Diels-Alder-Type Adducts: A Comparison of Sanggenon C with Sanggenon D.

Sanggenons C and D are two Diels-Alder-type adducts from Chinese crude drug Sang-bai-pi. Structurally, both sanggenons construct stereoisomers. In the study, they were comparatively determined using four antioxidant assays, including ferric ion reducing antioxidant power (FRAP) assay, Cu2+-reducing assay, 1,1-diphenyl-2-picryl-hydrazl (DPPH•)-scavenging assay, and 2,2′-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid radical (ABTS•+)-scavenging assay. Their Fe2+-binding reactions were explored using UV-Vis spectra. Finally, their cytoprotective effects were evaluated using flow cytometry. In electron transfer (ET)-based FRAP and Cu2+-reducing assays, sanggenon D was found to have lower IC50 values than sanggenon C; however, in multi-pathway-based DPPH•-scavenging and ABTS•+-scavenging assays, sanggenon C possessed lower IC50 values than sanggenon D. UV-Vis spectra suggested that sanggenon C generated a bathochromic-shift (286 nm → 302 nm) and displayed stronger UV absorption than sanggenon D. In flow cytometry, sanggenon C and sanggenon D, respectively, exhibited 31.1% and 42.0% early apoptosis-percentages towards oxidative-stressed mesenchymal stem cells (MSCs). In conclusion, both sanggenons may undergo multiple pathways (e.g., ET and Fe2+-binding) to protect MSCs against oxidative stress. In the mere ET aspect, sanggenon D possesses a higher level than sanggenon C, while in multi-pathway-based radical-scavenging, Fe2+-binding, and cytoprotection aspects, sanggenon C is more active than sanggenon D. These discrepancies can conclusively be attributed to the steric effect.

Oviductus ranae protein hydrolysate (ORPH) inhibits the growth, metastasis and glycolysis of HCC by targeting miR-491-5p/PKM2 axis

BackgroundOviductus Ranae (OR) is a valuable Chinese crude drug and has been reported to have a range of biological activities. Protein hydrolysate extracted from OR (ORPH) was previously found to have immune regulatory effect and anti-glioma activity. This study was aimed to investigate the effects of ORPH on hepatocellular carcinoma (HCC) progression. MethodsMTT, BrdU, colony formation and transwell assays were used to determine proliferation and mobility of HCC cells in vitro. Glucose consumption and lactate production assays were carried out to measure the glycolysis of HCC cells. The subcutaneous tumor model and lung metastasis model in nude mice were established to detect tumor growth and metastasis of HCC in vivo. The direct binding of miR-491-5p to 3’UTR of pyruvate kinase M2 (PKM2) was confirmed by luciferase reporter assay. ResultsIn vitro experiments showed that ORPH significantly inhibited proliferation, migration, invasion, epithelial-to-mesenchymal transition (EMT) and glycolysis of HCC cells. Moreover, ORPH treatment prominently suppressed HCC growth and metastasis in mice. We demonstrated that ORPH effectively decreased the expression of PKM2 in HCC cells. Forced expression of PKM2 abrogated the inhibitory effects of ORPH on HCC cells. Mechanically, ORPH reduced PKM2 expression in a post-transcriptional manner by up-regulating miR-491-5p. miR-491-5p exhibited a similar tumor suppressive effects with ORPH in HCC cells. Moreover, ORPH exerted its inhibitory effects on HCC cells through regulating miR-491-5p/PKM2 axis. Lastly, decreased miR-491-5p level and increased PKM2 expression were correlated with unfavorable clinical features and poor prognosis of HCC patients. ConclusionsIn all, this study reveals that ORPH inhibits the growth, metastasis and glycolysis of HCC cells by targeting miR-491-5p/PKM2 axis. ORPH may be a potential effective anti-tumor agent for HCC.

Epimagnolin A, a tetrahydrofurofuranoid lignan from Magnolia fargesii, reverses ABCB1-mediated drug resistance

BackgroundEpimagnolin A is an ingredient of the Chinese crude drug Shin-i, derived from the dried flower buds of Magnolia fargesii and Magnolia flos, which has been traditionally used for the treatment of allergic rhinitis and nasal congestion, empyema, and sinusitis. The pharmacokinetic activity of epimagnolin A remains to be evaluated. PurposeIn this study, we examined the possible interactions of epimagnolin A with human ATP-binding cassette (ABC) transporter ABCB1, a membrane protein vital in regulating the pharmacokinetics of drugs and xenobiotics. Study design/methodsThe interaction of epimagnolin A with ABCB1 was evaluated in calcein, ATPase, and MTT assays by using Flp-In-293/ABCB1 cells and purified ABCB1 and simulated in molecular docking studies. ResultsEpimagnolin A inhibited calcein export by Flp-In-293/ABCB1 cells in a concentration-dependent manner in a calcein assay. ATPase assay revealed a concentration-dependent stimulation of the ATPase activity of ABCB1 by epimagnolin A. Epimagnolin A also showed saturation kinetics in the relationship between the compound-stimulated ATPase activity and the compound concentration, suggesting Michaelis–Menten kinetics similar to those of the control drug, verapamil. Km and Vmax values were calculated from Hanes–Woolf plots of (compound concentration) × (compound-stimulated ATPase activity)−1 vs. (compound concentration); the Km of epimagnolin and verapamil was 42.9 ± 7.53 μM and 12.3 ± 4.79 μM, respectively, and the corresponding Vmax values were 156 ± 15.0 μM and 109 ± 3.18 μM. Molecular docking studies on human ABCB1 showed that epimagnolin A docked to the same binding pocket as verapamil, and 3-(4,5-dimethyl-2-thiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays showed that the sensitivities of Flp-In-293/ABCB1 cells against anti-cancer drugs were enhanced upon exposure to 10 μM epimagnolin A. ConclusionThese results strongly suggest that epimagnolin A affects the transport activity of ABCB1 as a substrate.

Taxonomic examination of longgu (Fossilia Ossis Mastodi, "dragon bone") and a related crude drug, longchi (Dens Draconis, "dragon tooth"), from Japanese and Chinese crude drug markets.

Longgu ("dragon bone," Ryu-kotsu, Fossilia Ossis Mastodi, or Os Draconis) is the only fossil crude drug listed in the Japanese Pharmacopoeia. All longgu in the current Japanese market is imported from China, where its resources are being depleted. Therefore, effective countermeasures are urgently needed to prevent resource depletion. One possible solution is the development of a substitute made from bones of contemporary animals that are closely related to the original animal source of the current longgu. However, no research has been conducted on the original animal source of longgu, except for a report on the longgu specimens present in the Shosoin Repository. Taxonomic examination was performed on the fossil specimens related to longgu which are owned by the Museum of Osaka University, Japan. In total, 20,939 fossil fragments were examined, of which 20,886 were mammalian fossils, and 246 of these fossils were classified into nine families. The longgu specimens from the Japanese market belonged to a relatively smaller variety of taxa than those from the Chinese market. Despite the variety of taxa in longgu, medical doctors using Kampo preparations with longgu have not reported any problems due to the presence of impurities in the original animal source. These results suggest that the effect of longgu is independent of its origin as long as it is closely related to the origin of the current longgu. Thus, despite the considerable effects of fossilization, our results could help in developing an optimal substitute for longgu.

New Homoisoflavanes, a New Alkaloid and Spirostane Steroids from the Roots of Herreria montevidensis Klotzsch ex Griseb. (Herreriaceae).

The roots of the South American vine Herreria montevidensis Klotzsch ex Griseb. (Herreriaceae) are used in traditional medicine by several Amerindian groups of the Paraguayan Chaco. Little is known on the chemistry of the plant, despite its widespread use across the South American Chaco. From the ethyl acetate/methanol 1:1 extract of the roots, four new and one known homoisoflavanoid, two flavan derivatives, a stilbene, a new alkaloid, and three new and four known spirostane steroids were isolated. The corresponding structures were elucidated by spectroscopic and spectrometric means. The homoisoflavonoids of the plant are related to compounds isolated from the Dracaenaceae (formerly Agavaceae) sources of the Chinese crude drug Dragon’s Blood. The new alkaloid is a novel skeleton that can be used as a chemical marker of Herreria. The spirostane steroids suggest chemotaxonomic relations with the Liliales. This is the first comprehensive report on the chemistry of a South American Herreria species.

金果榄药材的UPLC指纹图谱研究

目的：建立金果榄药材的UPLC指纹图谱，为金果榄药材的质量控制提供依据。方法：采用超高效液相色谱法。色谱条件：色谱柱：Acquity UPLC®BEN C18 (2.1 × 100 mm，1.7 µm)；流动相A：乙腈；流动相B：0.3%磷酸溶液；流速：0.25 ml/min；检测波长：225 nm；柱温：25℃。结果：建立了10批金果榄药材的指纹图谱。金果榄药材有7个共有峰，多数峰可以达到较好分离，具有较高的相似度。结论：建立的超高效液相指纹图谱有较好的精密度、重复性和稳定性，可作为金果榄质量评价参考。 Objective: To establish UPLC fingerprint of Tinospora sagittata and T. capillipes which is expected to provide standards for the quality control and identification of the Chinese crude drug. Method: The UPLC method was used. Chromatography conditions were Acquity UPLC®BEN C18 (2.1 × 100 mm, 1.7 µm) column with gradient mobile phase of acetonitrile and 0.3% phosphate acid. UV detection wavelength was 225 nm, the flow rate was 0.25 ml/min and the column temperature was 25˚C. Result: UPLC fingerprints of 10 samples of Tinospora sagittata and T. capillipes were established. 7 mutual peaks were selected as the fingerprint peaks in the samples. Among the obtained fingerprints, most of the detected peaks were separated effectively. 10 samples had higher simi-larities. Conclusion: The established UPLC fingerprint has desirable accuracy, repeatability and sta-bility, which can be used as reference for the quality control of Tinospora sagittata and T. capillipes.

Advances in study of anticancer properties of Allii Macrostemonis Bulbus

A commonly used Chinese crude drug Allii Macrostemonis Bulbus has been shown to possess good anticancer activities and related properties such as antioxidation, nitrite scavenging, nitrosamine synthesis blocking and immune enhancement, and has been widely used as an effective auxiliary drug in the treatment of some malignant tumors. This paper systematically reviews the advances in the study of anticancer-related activities of Allii Macrostemonis Bulbus's various components such as raw juice, extracts, saponins, volatile oil, polysaccharides, nitrogen compounds, etc.

Study on Fingerprint of ZheRong Radix Pseudostellariae by HPLC

To establish HPLC fingerprint of ZheRong Radix pseudostellariae and the evidence was provided for quality control and identification of the Chinese crude drug. The HPLC method was used,chromatography conditions were Agilent Eclipse XDB-C18( 4. 6 mm ×250 mm,5 μm) column with gradient mobile phase of acetonitrile-water,The detection wavelength was 203 nm and the col-umn temperature was 30 °C with the flow rate of 1. 0 mL/min,the sample injection was 20μL. HPLC fingerprints of 32 sam-ples of ZheRong Radix pseudostellariae were established. 9 common peaks were selected as the fingerprint peaks in all samples. Among the obtained fingerprints,most of the detected peaks were separated effectively. 32 samples had high similarities. The established HPLC fingerprint has desirable accuracy,repeatability and stability,which can be used for one of the quality control methods of ZheRong Radix pseudostellariae.

Collection, Identification, Phytochemical analysis and Phyto toxicity test of Wood inhabiting Fungi Ganoderma lucidum (Curt.Fr.)P.Karst.

Plan: In this study 10 strains of mushroom Ganoderma lucidum were collected from various places and decided to screen their phytotoxicity activity and phytochemical constitution. Preface: The popular mushroom Ganoderma lucidum (Reishi) was a bracket fungus and has been widely used for the promotion of health and longevity in Asian countries. The dried powder of G.lucidum was popular as a cancer chemotherapy agent in ancient China. G.lucidum clearly demonstrates anticancer activity in experiments with cancer cells and has possible therapeutic potential as a dietary supplement or alternative therapy for breast and prostate cancer. And also the fruiting bodies of Ganoderma lucidum (Polyporaceae) are a well known Chinese crude drug identified as a phytotoxic agent. H owever, since G.lucidum was available from different sources, it is advisable to test its biological activity. Methodology: G.lucidum strains collected was isolated and identified with potato dextrose agar medium. Then the secondary metabolites were qualitatively studied through phytochemical analysis. Outcome: Significant root length inhibition was observed at 100ppm and 200ppm. Similarly seed germination was also significantly inhibited at the concentration 100ppm and 200ppm extracts. The mean data of root length inhibition by ethanol extract in 100ppm was 0.393 and 0.208 in 200ppm. Likewise the mean data of seed germination inhibition by ethanol extract in 100ppm was 15 and 0.208 in 200ppm. Overall results supported that Ganoderma lucidum clearly suppo rted the wider medicinal uses and established its anticancer activity.

Immunostimulatory activity of protein hydrolysate from oviductus ranae on macrophage in vitro.

Oviductus Ranae is the dry oviduct of Rana chensinensis, which is also called R. chensinensis oil. Oviductus Ranae is a valuable Chinese crude drug and is recorded in the Pharmacopoeia of the People's Republic of China. The aim of this study was to investigate the immunostimulatory activity of protein hydrolysate of Oviductus Ranae (ORPH) and to assess its possible mechanism. Immunomodulatory activity of ORPH was examined in murine macrophage RAW 264.7 cells. The effect of ORPH on the phagocytic activity of macrophages was determined by the neutral red uptake assay. After treatment with ORPH, NO production levels in the culture supernatant were investigated by Griess assay. The mRNA and protein expressions of inducible nitric oxide synthase (iNOS) were detected by RT-PCR and Western blotting. The production of TNF-α, IL-1β, and IL-6 after treatment with ORPH was measured using ELISA assay. In addition, NF-κB levels were also investigated by Western blot. The results showed that ORPH enhanced the phagocytosis of macrophage, increased productions of TNF-α, IL-1β, IL-6, and NO in RAW 264.7 cells, and upregulated the mRNA and protein expression of iNOS. Besides, NF-κB, levels in RAW 264.7 cells were elevated after ORPH treatment. These findings suggested that ORPH might stimulate macrophage activities by activating the NF-κB pathway.

New bencaological studies of traditional Chinese medicine after name "Doukou"

Lots of traditional Chinese medicine (TCM) shares the same Chinese common names "Doukou". Because of similar Chinese names, appearances, functions and original plants, there are still no compromise on the original plants of "Doukou" up to now. Moreover, "Doukou" referred to more than one source of species, that is, it might refer to the Chinese crude drug derived from different plants during different historical periods. In order to identify the original botanical plants of "Doukou" during different historical periods and the relationship between these "Doukou", new bencaological studies of TCM under the name "Doukou" were carried out, which included the studies on literal description, image description, market investigation and systematic botanical research. A suggestion was made to change the Chinese name "Doukou" (Amomi Fructus Rotundus) to "Baidoukou".

Anti-Austerity Agents from Rhizoma et Radix Notopterygii (Qianghuo)

During a search for potent anticancer agents from natural products based on an anti-austerity strategy, we found that a CHCl3 extract of Rhizoma et Radix Notopterygii (Qianghuo), a Chinese crude drug, exhibited strong cytotoxicity against PANC-1 human pancreatic cancer cells, with a PC₅₀ value of 17.5 µg/mL. Further fractionation and purification of this bioactive extract led to the isolation of 19 known compounds. The in vitro preferential cytotoxicity of the isolates was evaluated against two human pancreatic cancer cell lines, PANC-1 and PSN-1. Among the compounds isolated, ostruthin displayed the most potent activity against both PANC-1 (PC₅₀, 7.2 µM) and PSN-1 (PC₅₀, 7.8 µM) cells in nutrient-deprived medium (NDM) and may have induced necrotic nutrient-deprived PANC-1 cell death.

Research on Quantitative Prediction of Total Ash in Pollen Typhae by NIRS

Near infrared spectroscopy is used for establishing a rapid and non-destructive analytical method to determine the quantitative of Total Ash in Pollen Typhae, a traditional chinese crude drug. Partial least squares regression (PLSR) was used for building the quantitative prediction model of Pollen Typhae different calibration methods were compared with spectral pretreated methods, wave-number selection and factors of NIRS were discussed in detail. The first derivative and multiplicative scatter correction method was used to predict total ash to correlation coefficients (R), root mean square error of calibration (RMESECV) residual prediction deviation (RPD) and offset obtained by PLSR model were 0.9994, 0.293, 29.4 and 0.013 respectively. The standard deviation was less than 8.2% by the total ash prediction model in the 8 batches of Pollen Typhae from 6% to 27.3%. The predictive result shows the method of this research is rapid, non-destructive and credible, which can be applied to the rapid predict the total ash of Pollen Typhae.

Authentication of Chinese Crude Drug Gecko by DNA Barcoding

Gekko gecko, an animal used as a valued traditional Chinese medicine, has been widely used for over 2000 years. Due to localized habitat destruction, the amount of G. gecko has dramatically decreased in recent years. As a result, more and more adulterants have been detected in the traditional medicine, which has resulted in a chaotic market. Therefore, a correct identification method is badly needed. In this study, we employed a new molecular method of DNA barcoding for discriminating gecko from its adulterants. Fifty-seven specimens of gecko and its adulterants were collected as test samples. The full-barcode and mini-barcode sequences of these specimens were separately amplified and sequenced separately. Together with other published barcode sequences, we detected that the intra-specific sequence diversity was far lower than the inter-specific diversity in G. gecko and its adulterants (3% compared with 35% in full-length barcode; 4% compared with 33.5% in mini-barcode). These results showed that both the full-length and mini-barcodes were effective for identifying gecko, which suggested that the DNA barcode could be an effective and powerful tool for identifying the Chinese crude drug gecko.

On the curing of the Chinese crude drug and the Chinese medicinal alcoholic drinks

On the curing of the Chinese crude drug and the Chinese medicinal alcoholic drinks

Isolation of a renal function-facilitating constituent from the Oriental drug, salviae miltiorrhizae radix.

An attempt was made to isolate the active component which exhibits an improving effect on renal function from Salviae Miltiorrhazae Radix (Chinese crude drug). Systematic isolation from aqueous extract of Salviae Miltiorrhizae Radix was carried out, and Compound 1 was found to be more effective than any of the other constituents in improving renal functional parameters; that is, a marked reduction of glomerular filtration rate following adenine ingestion was improved by administration of this substance. The renal plasma flow and renal blood flow were also increased in renal failure rats. On the basis of chemical and spectroscopic data, Compound 1 was shown to be identical with magnesium lithospermate B.

ChemInform Abstract: Astertarone B, a Hydroxy-Triterpenoid Ketone from the Roots of Aster tataricus L.

The roots of Aster (A.) tataricus L. (Compositae) are used as the Chinese crude drug Asteris Radix (Japanese name: shion) and are combined in various traditional Chinese prescriptions for use as cough medicines, expectorants and diuretics 1) . Constituents of the root extracts have been investigated, and the presence of a triterpenoid ketone, shionone (shion-21-en-3-one), 2—4) seven triterpenoid glycosides, aster saponins A—G, 5—7) and three monoterpenoid glycosides, shionosides A—C, 7,8) have been reported. Our recent study on the methanol extract of A. tataricus roots has led to the isolation and structural elucidation of a new triterpenoid ketone, astertarone A (D: A-friedoeuph-21-en-3-one), along with epishionol (shion-21-en-3b -ol). 9) In this paper, we report the isolation and characterization of yet another new triterpenoid ketone, designated astertarone B (1), in addition to spinasterone [(22E)-5a-stigmasta-7,22-dien-3-one], from the extract. Column chromatography of the hexane soluble portion of the methanol extract of A. tataricus roots on silica gel afforded a triterpenoid ketone fraction. Crystallization of the ketone fraction from acetone‐methanol, followed by preparative HPLC of the filtrate portion, eventually yielded compound 1 and spinasterone. Identification of spinasterone was performed by spectral comparison with an authentic compound. Compound 1, with a molecular formula of C30H50O2 determined from its high-resolution mass spectrum (HR-MS) (M 1 , m/z 442.3836), gave IR absorptions at 3473 (hydroxyl group), 1712 (ketone), and 833 cm 21 (trisubstituted double bond). Compound 1 displayed two olefinic methyl singlets at d 1.61 and 1.69, with an olefinic methine signal at d 5.10 (br t, J57.0 Hz), which correspond to one isopropylidene group, and six tertiary methyl singlets at d 0.82, 0.89, 0.90,