Bioactive-guided isolation and identification of oligostilbenes as anti-rheumatoid arthritis constituents from the roots of Caragana stenophylla.

ETHNOPHARMACOLOGICAL RELEVANCE: The roots of Caragana stenophylla have been used as folk medicine due to the functions of activating blood, diuresis, analgesic and tonicity, especially in treating rheumatoid arthritis and hypertension. However, the anti-rheumatoid arthritis mechanisms and bioactive ingredients have not previously been fully investigated. AIM OF THE STUDY: The aim of this study was to assess the anti-rheumatoid arthritis effects of the roots of Caragana stenophylla ethanol extract (EC), elucidate its mechanism of action and identify its active substances. MATERIALS AND METHODS: Anti-rheumatoid arthritis activity of EC was assessed using type II-collagen induced arthritis in rats. Arthritis severity was evaluated by foot paw volume, arthritis index, joint swelling degree and histopathology. The serum inflammatory cytokines and matrix metalloproteinases (MMPs) were also detected by immunohistochemical analysis. In addition, the protein expression of IκB, p-IκB, iNOS and COX-2 was analyzed by western blot. RAW 264.7 macrophage cells were employed to assess the anti-inflammatory effects of fractions and compounds in vitro. UPLC-Q-TOF-MS was adopted to appraise the ingredients of the active fraction of the roots of C. stenophylla. Furthermore, various chromatographic techniques and spectroscopic methods were used for isolation and structure elucidation of compounds. RESULTS: The results showed that EC could reduce type II collagen-induced rheumatoid arthritis model arthritic score and histopathology markedly at dose of 240 mg/kg. Besides, EC could suppress the levels of pro-inflammatory cytokines (IL-1ß, IL-6, IL-8, IL-17, and TNF-α) and matrix metalloproteinases (MMP-3, MMP-9), and the expression levels of COX-2, p-IκB and iNOS also were declined. While, the levels of IL-10 and IκB were increased. The ethyl acetate fraction exhibited potent inhibitory effects against nitric oxide production in RAW 264.7 macrophage cells. Eleven main components including 1 flavonoid and 10 oligostilbenes from active fraction were isolated by mass directed chromatographic techniques. Their structures were determined on the basis of various spectroscopic methods and by comparison with the published NMR data. CONCLUSION: The roots of C. stenophylla attenuated arthritis severity, restored serum cytokine imbalances by regulating NF-κB signaling pathway in type II collagen-induced rheumatoid arthritis model. Oligostilbenes were essential ingredients in ethyl acetate extract of C. stenophylla roots. Stilbenes and flavonoids should be responsible for its anti-rheumatoid arthritis activities.

Four New Lignans from Kadsura Interior and Their Bioactivity.

A phytochemical investigation of the stems of Kadsura interior has led to an isolation of four new lignans, named kadsutherin E⁻H (1⁻4), together with two known lignans (5⁻6). The structures of the four new compounds were established on the basis of comprehensive spectroscopic analyses. Compounds 1⁻6 exhibited inhibition against adenosine diphosphate (ADP) induced platelet aggregation. Among the isolated compounds, kadsutherin F (2) showed the strongest anti-platelet aggregation activity with an inhibition of 49.47%.

[Chemical constituents from roots of Caragana stenophylla and their anti-tumor activities].

Fourteen compounds were isolated from the 80% ethanol extract of Caragana stenophylla root, by using a combination of various chromatographic approaches, including silica gel sephadex LH-20 column chromatography, and preparative HPLC. On the basis of their physical and chemical properties and spectroscopic data, their structures were elucidated as 2-(4-hydroxy-3-methoxy lphenyl)-3-methoxyl benzofuran-6-ol (1), mucodianin C (2), isopterofuran (3), formononetin (4), afromosin (5), calycosin (6), acacetin (7), 3-O-methylkaempferol (8), liquiritigenin (9), isoliquiritigenin (10), variabilin (11), resveratrol (12), zhebeiresinol (13), and 2, 3-dicarboxy-6, 7-dihydroxy-1-(3', 4'-dihydroxy)-phenyl-1, 2-dihydronaphthalen (14). Compound 1 is a new benzofuran derivative, named as mucodianin S; compounds 2, 3, 11, 13, 14 were isolated from the genus Caragana for the first time, and compounds 4-10 were firstly isolated from Caragana stenophylla. MTT assay was used to determine their cytotoxicity of the isolated compounds against human tumor cell lines, and 2 showed cytotoxicity against human hepato cellular cancer (HepG2) and human cervical (HeLa) lines, with IC50 values of (16.18±0.95), (3.75±0.08) µmol•L ⁻¹, respectively.

[Projection of potential geographic distribution of Apocynum venetum under climate change in northern China].

Apocynum venetum belongs to apocynaceae and is a perennial medicinal plant, its stem is an important textile raw materials. The projection of potential geographic distribution of A. venetum has an important significance for the protection and sustainable utilization of the plant. This study was conducted to determine the potential geographic distribution of A. venetum and to project how climate change would affect its geographic distribution. The projection geographic distribution of A. venetum under current bioclimatic conditions in northern China was simulated using MaxEnt software based on species presence data at 44 locations and 19 bioclimatic parameters. The future distributions of A. venetum were also projected in 2050 and 2070 under the climate change scenarios of RCP2.6 and RCP8.5 described in 5th Assessment Report of the Intergovernmental Panel on Climate Change (IPCC). The result showed that min air temperature of the coldest month, annual mean air temperature, precipitation of the coldest quarter and mean air temperature of the wettest quarter dominated the geographic distribution of A. venetum. Under current climate, the suitable habitats of A. venetum is 11.94% in China, the suitable habitats are mainly located in the middle of Xinjiang, in the northern part of Gansu, in the southern part of Neimeng, in the northern part of Ningxia, in the middle and northern part of Shaanxi, in the southern part of Shanxi, in the middle and northern part of Henan, in the middle and southern part of Hebei, Shandong, Tianjin, in the southern part of Liaoning and part of Beijing. From 2050 to 2070, the model outputs indicated that the suitable habitats of A. venetum would decrease under the climate change scenarios of RCP2.6 and RCP8.5.

Natural neuro-inflammatory inhibitors from Caragana turfanensis.

Because of the critical role of over-activated microglia in the progress of neurodegenerative diseases, it has been selected as a potential therapeutic target for drug discovery. In order to find natural neuroinflammatory inhibitors, we carried out a bioactivity-oriented phytochemical research of Caragana turfanensis Kom. (Krassn.), which is a folk medicine widely distributed in Xinjiang. As a result, a new coumarin lactone caraganolide A (1) and 35 known components were characterized from the effective extract of C. turfanensis. Furthermore, their anti-neuroinflammatory effects were evaluated in LPS-induced BV2 microglial cells using Griess assay to determine the release of nitric oxide (NO). Compounds 1, 2, 4-6, 9, 13-15, 20, 29 and 30 exhibited significant inhibitory activities and no obvious cytotoxicities were observed at their effective concentrations. It is noteworthy, the new compound caraganolide A (1) (IC50 1.01±1.57µM) and 3',7,8-trihydroxy-4'-methoxyisoflavone (5) (IC506.87±2.23µM) exhibited more excellent action than that of positive control minocycline (IC50 9.07±0.86µM).

Natural potential neuroinflammatory inhibitors from Alhagi sparsifolia Shap.

Neuroinflammation is a key contributor to neuronal damage in neurodegenerative diseases. In our previous work on natural effective neuroinflammatory inhibitors, Alhagi sparsifolia Shap. (Leguminosae), a folk medicine widely distributed in Xinjiang, attracted our attention because of its significant anti-neuroinflammatory effect. Therefore, further investigation of the bioactive material basis was carried out. As a result, 33 major components were characterized and identified by chromatographic and spectral methods, respectively. Furthermore, the anti-neuroinflammatory effects of the extract and purified constituents were evaluated in LPS-induced N9 cells in vitro. The results displayed that compounds 1, 2, 3, 5, 6, 8, 11, 15, 16, 17, 22, 23, 25, 26, 28, 30, 33 could exhibit significant inhibitory activities without obvious cytotoxicities at their effective concentrations. Especially, isorhamnetin (1) (IC50 17.87µM), quercetin (2) (10.22µM), 3',7-dihydroxyl-4'-methoxylisoflavone (5) (17.43µM), 3',7-dihydroxyl-4',6-dimethoxylisoflavone (6) (11.21µM), syringgaresinol (16) (2.68µM), bombasinol A (17) (7.61µM), aurantiamide (23) (14.91µM) and 1,3,3,4-tetramethyl cyclopentene (33) (2.63µM) showed much stronger inhibiting effect than that of the positive control minocycline (19.89µM). Therefore, the effective compositions might be responsible for the significant neuroinflammation inhibitory activities exhibited by the herb. Moreover, compounds 16 and 33 could be good leading compounds for the development of potential therapeutic agents against neurodegenerative diseases.

Flavonoids from Caragana pruinosa roots.

A new pterocarpan derivative, pruinosanone D (1), a new isoflavonoid, pruinosanone E (2), and a new chalcone, pruinosanone F (3), were isolated from Caragana pruinosa roots, along with four known analogues (4-7), identified as 2,4-dihydroxy-3'-methoxy-4'-ethoxychalcone, 7,4-dihydroxyflavanone, butin and scutellaprostin C, respectively. Their structures were elucidated by detailed analyses of NMR, IR, and MS data. The ability of the isolated compounds to prevent nitric oxide (NO) production by LPS-stimulated RAW 264.7 macrophages was also studied. Compound 1 were among the most potent NO production inhibitor, with IC50 value of 0.62µM.

Therapeutic effects of Caragana pruinosa Kom. roots extract on type II collagen-induced arthritis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Caragana pruinosa Kom. is a deciduous shrub belonging to the genus of Caragana (Leguminosae), and Caragana plants exhibit a wide range of interesting pharmacological properties including anti-inflammatory, analgesic, and anti-arthritis activity, etc. AIM OF THE STUDY: This study was aimed to investigate the anti-arthritic effect of 80% EtOH extract from the roots of C. pruinosa (ERCP) on arthritis and explore the potential pharmacological mechanism. MATERIALS AND METHODS: After collagen induced arthritis (CIA) were established in rats, the animals were orally administered with ERCP (130, 260 and 520mg/kg) for 30 days. During the treatment, the rats' body weights, arthritis indices and paw volumes were measured every 5 days. Subsequently, rats' blood samples were collected to determine TNF-α, IL-1ß, IL-6, IL-10, and C-reactive protein (CRP) contents in serum. Then, rats were sacrificed and the hind paws and knee joints were collected for histopathological examination. RESULTS: Our results indicated that ERCP significantly suppressed the inflammatory reactions and destructions in joints and synovial tissues. ERCP inhibited the paw swelling and arthritis index in CIA rats. Additionally, it decreased the levels of pro-inflammatory cytokines (TNF-α, IL-1ß and IL-6) and CRP, whereas increased that of IL-10. CONCLUSION: Our results suggested ERCP has significant anti-arthritic effect on CIA rats, and the pharmacological mechanisms are related to the down-regulation of TNF-α, IL-1ß, IL-6 and CRP and the up-regulation of IL-10.

Diarylpentanol constituents from the aerial part of Stelleropsis tianschanica.

Five diarylpentanol derivatives including two new compounds stellerasme A (1), stellerasme B (2) were isolated from the aerial parts of Stelleropsis tianschanica. Their structures were elucidated by various spectroscopic techniques (UV, IR, MS, CD, 1D and 2D NMR). All compounds were evaluated for their cytotoxicity activity against HeLa and KB cell lines, and compound 1 showed selective activities against HeLa cell line with an IC50 value of 7.4 µM.

Simultaneous Quantitative and Chemical Fingerprint Analysis of Receptaculum Nelumbinis Based on HPLC-DAD-MS Combined with Chemometrics.

A rapid and sensitive method based on HPLC-DAD-MS was developed for quantitative analysis of two flavonoids and chemical fingerprint analysis to evaluate the quality of Receptaculum Nelumbinis. The analysis was conducted on a Poroshell 120 C18 column (100 × 4.6 mm, 2.7 µm) with 0.2% formic acid buffer solution and methanol as mobile phases with gradient elution. This method displayed good linearity with R(2) at >0.9999 and limits of quantity <0.37 µg mL(-1). Relative standard deviation values for intra- and interday precision were <0.82 and 1.03%, respectively. The mean recovery of hyperoside was 95.54% and of isoquercitrin was 92.10%. Hyperoside and isoquercitrin were determined simultaneously, and 12 peaks in the chemical fingerprint were identified. The chemometric methods, including similarity analysis, hierarchical clustering analysis and principal component analysis, were applied to distinguish 11 batches of Receptaculum Nelumbinis samples. The above results could validate each other and successfully divide these samples into two groups. Moreover, hyperoside and isoquercitrin could be selected as chemical markers to evaluate the quality of Receptaculum Nelumbinis from different localities. This study demonstrated that the developed method was a powerful and beneficial tool to carry out the quality control of Receptaculum Nelumbinis.

Systematic chemical analysis of flavonoids in the Nelumbinis stamen.

The stamen of lotus, known as Nelumbinis stamen, has been used as the folk medicine and functional food for a long time, which showed good activities of anti-ulcer, anti-thrombosis, analgesic, anti-diarrhea, strengthen uterine contraction. The bioactivities of Nelumbinis stamen were attributed to the existence of flavonoids, its characteristic chemical constituents. A reliable method for comprehensive chemical analysis of flavonoids in Nelumbinis stamen by HPLC-DAD-MS was developed for the first time. The extraction protocol of flavonoids from Nelumbinis stamen was optimized by an orthogonal design. The chromatographic conditions were optimized, which exhibited similar level than that of the UHPLC platform allowing target compound identification in a shorter time with little solvent consumption. Moreover, similarity analysis, hierarchical clustering analysis and principal components analysis were successfully applied to demonstrate the variability of these Nelumbinis stamen samples.

[Evaluation of high-resolution images application for wild medicinal plants macro monitoring: a case of Apocynum].

To investigate the resources of medicinal plant, such as wild Apocynum, supervised classification based on Principal Component Analysis (PCA) and texture feature were used to monitor wild medicinal plants from image captured by ZY-3 and World-view-2 and compare which satellite Image are more appropriate to monitor the wild medicinal plants. The research results shows that: for more complex growth conditions wild medicinal plants Apocynum, high-resolution images Worldview-2 is more suitable for its remote identification, the low-resolution satellite ZY-3 can only recognizes the wild medicinal plants which distributed intensively. If the study target distribution is more intensive and larger scale, and cultivated type medicinal plants, the use of satellite ZY-3 in low resolution remote sensing data to identify the target can be a good choice, it is not necessary to buy high-resolution data, in order to avoid waste of expenditure, for the scattered distribution, the high-resolution satellite imagery data may be indispensable to identify targets.

[Research on spectral reflectance characteristics for Glycyrrhizae Radix].

In order to study the spectral reflectance differences of Glycyrrhizae Radix under different growth conditions and lay the foundation for quantitative monitoring of Glycyrrhizae Radix remote sensing images, spectra of Glycyrrhiza species under different growth period and different varieties and different regions were measured by a portable spectrometer. The results showed that the reflectivity of annual G. uralensis was obviously higher than that of the two years plant in the visible light band own to the contents of crown layer chlorophyll. The reflectivity of two years G. pallidiflora was higher than that of G. uralensis in the near infrared band own to the leaf area index and the content of leaf water. The red edge spectrum of annual plant fluctuated largely than that of two years plant due to vegetation coverage and leaf area index. G. pallidiflora grew well than G. uralensis. Under different regions of the Glycyrrhiza species, spectral data analysis showed that within a certain range, the average annual precipitation and average annual evaporation were the major factors to affect the differences of Glycyrrhiza species spectral data under different regions owe to the leaf water content, the higher leaf water content, the lower spectral reflectance. The principal component analysis and continuum-removed method of the spectral data under different regions found that, within a certain range, the average annual precipitation and average annual evaporation were the major factors caused by the differences of Glycyrrhiza species spectral data under the different regions, Glycyrrhiza species spectral similarity related to the spatial distance.

Genus Kadsura, a good source with considerable characteristic chemical constituents and potential bioactivities.

In China, the plants of genus Kadsura had been used as the folk medicines for a long time and showed good effect of activating blood and dissolving stasis, promoting qi circulation to relieve pain, dispelling wind and eliminating dampness. The bioactivities of genus Kadsura were attributed to the existence of its characteristic chemical constituents. This review systematically summarized the traditional efficacy and medicinal application of genus Kadsura in China, chemical constituents and bioactivities of the plants of genus Kadsura. And, lignans and triterpenoids were the main bioactive constituents, which exhibited good anti-HIV, anti-tumor, anti-hepatitis, anti-oxidant, anti-platelet aggregation activities and neuroprotective effect etc. Moreover, some structure-activity relationships mining would greatly enrich the opportunity of finding new and promising lead compounds and promote the reasonable development and utilization of the plants of genus Kadsura.

Acylated phenylethanoid glycosides, echinacoside and acteoside from Cistanche tubulosa, improve glucose tolerance in mice.

Acylated phenylethanoid glycosides, echinacoside (1) and acteoside (2), principal constituents in stems of Cistanche tubulosa (Orobanchaceae), inhibited the increase in postprandial blood glucose levels in starch-loaded mice at doses of 250-500 mg/kg p.o. These compounds (1 and 2) also significantly improved glucose tolerance in starch-loaded mice after 2 weeks of continuous administration at doses of 125 and/or 250 mg/kg/day p.o. without producing significant changes in body weight or food intake. In addition, several constituents from C. tubulosa, including 1 (IC50 = 3.1 µM), 2 (1.2 µM), isoacteoside (3, 4.6 µM), 2'-acetylacteoside (4, 0.071 µM), tubulosides A (5, 8.8 µM) and B (9, 4.0 µM), syringalide A 3-O-α-L-rhamnopyranoside (10, 1.1 µM), campneoside I (13, 0.53 µM), and kankanoside J1 (14, 9.3 µM), demonstrated potent rat lens aldose reductase inhibitory activity. In particular, the potency of compound 4 was similar to that of epalrestat (0.072 µM), a clinical aldose reductase inhibitor.

Rapid discrimination of Herba Cistanches by multi-step infrared macro-fingerprinting combined with soft independent modeling of class analogy (SIMCA).

Herba Cistanche, an important Chinese herbal medicine, has common four species, Cistanche deserticola (CD), Cistanche tubulosa (CT), Cistanche salsa (CS) and Cistanche sinensis (CSN) which have been frequent mixed used. To clarify the sources of Herba Cistanches and ensure the clinical efficacy and safety, a method combing IR macro-fingerprinting with statistical pattern recognition was developed to analyze and discriminate the four species of Herba Cistanche. By comparing FT-IR, second derivative spectral fingerprints via group-peak matching, the similarity to CD and total saccharides (TS) followed an increasing sequence, CTCSN>CS>CD. Characteristic fingerprints of their 2D-IR correlation spectra in 1750-1000 cm(-1) have confirmed the above findings in a more intuitive way. In terms of sources for phenylethanoid glycosides (PhGs), CT can be an ideal alternative species. However, in terms of using them as a whole, more pharmacological study should be conducted due to the different ratios of their chemical constituents, which is also applicable to CSN and CS. Moreover, the four species (179 samples) has been objectively classified by SIMCA based on IR macro-fingerprints.

[Measurement of sown area of safflower based on PCA and texture features classification and remote sensing imagery].

To improve accuracy of estimation in planted safflower acreage,we selected agricultural area in Yumin County, Xinjiang as the study area. There safflower was concentrated planted. Supervised classification based on Principal Component Analysis (PCA) and texture feature were used to obtain the safflower acreage from image captured by ZY-3. The classification result was compared with only spectral feature and spectral feature with texture feature. The research result shows that this method can effectively solve the problem of low accuracy and fracture classification result in single data source classification. The overall accuracy is 87.519 1%, which increases by 7.117 2% compared with single data source classification. Therefore, the classification method based on PCA and texture features can be adapted to RS image classification and estimate the acreage of safflower. This study provides a feasible solution for estimation of planted safflower acreage by image captured by ZY-3 satellite.

Flavonol glycosides, nigelflavonosides A-F from the whole plant of Nigella glandulifera (Ranunculaceae).

Six new flavonol glycosides, nigelflavonosides A-F (1-6), together with a known compound (7) were isolated from the whole plant of Nigella glandulifera Freyn et Sint (Ranunculaceae). Structure elucidation, especially the localization of the glycosyl or acetyl groups, and complete (1)H- and (13)C-NMR assignments of these compounds were carried out using one- and two-dimensional NMR measurements, including (1)H- and (13)C-NMR, (1)H-(1)H COSY, TOCSY, HMQC, HMBC and NOESY, in addition to HRESI-TOF-MS experiments.

[Speciation analysis of harmful elements in Rhei Radix et Rhizoma, Chrysanthemi Flos].

OBJECTIVE: To investigate the state and distribution of Pb, Cd, Hg, As, Cu in Rhei Radix et Rhizoma and Chrysanthemi Flos. METHOD: The samples were prepared by modified Tessier sequential extraction; The elements of Cu, Pb, and Cd in the samples were analyzed by atomic absorption spectrometry (AAS), while Hg, and As were analyzed by atomic fluorescence (AFS). RESULT: Cu,Pb,Cd,Hg,As in Chrysanthemi Flos were 12.806, 10.478, 0.436, 0.231, 1.531 mg x kg(-1), respectively. Cu, Pb, Hg in Chrysanthemi Flos mainly existed in residual and organic states; Cd was priority to ion exchange state; the residual state was the main form of As, and ion exchange state and water soluble state also had a large proportion. Cu, Pb, Cd, Hg, As in Rhei Radix et Rhizoma were 10.530, 4.926, 0.478, 0.260, 0.750 mg x kg(-1), respectively. Cu, Pb and Hg of residual state were the highest ratio; Cd and As mainly existed in ion exchange state. CONCLUSION: The experiment results showed that sequential extraction can be applied in speciation analysis of the harmful elements of traditional Chinese medicine. The method can speciate the state and distribution of harmful elements and provide the information of harmful elements. It will provide reference to the production of Chinese traditional patent medicine and herb extracts, processing of medicine herbs, the development of new drugs and safety evaluation of traditional Chinese medicine.

Sesquiterpene lactones from Saussurea involucrata.

Bioassay-directed separation of the ethyl acetate extract from the aerial parts of Saussurea involucrata (Kar. et Kir. ex Maxim) led to the isolation of three new sesquiterpene lactones sausinlactones A (1), B (2), C (3) and six known ones (4-9). The structures were established by spectroscopic data (UV, IR, HRESIMS, 1D and 2D NMR). CD technique was also employed to determine the absolute configurations of new compounds 1-3. The anti-inflammatory activities of compounds 1-9 and antitumor activities of new compounds 1-3 were tested. The results presented that compounds 5 and 6 were responsible for anti-inflammatory activities, and compounds 1 and 2 showed significant cytotoxic activities against A549 cells.