Rapid Screening of Active Components with an Osteoclastic Inhibitory Effect in Herba epimedii Using Quantitative Pattern-Activity Relationships Based on Joint-Action Models.

Screening of bioactive components is important for modernization and quality control of herbal medicines, while the traditional bioassay-guided phytochemical approach is time-consuming and laborious. The presented study proposes a strategy for rapid screening of active components from herbal medicines. As a case study, the quantitative pattern-activity relationship (QPAR) between compounds and the osteoclastic inhibitory effect of Herba epimedii, a widely used herbal medicine in China, were investigated based on joint models. For model construction, standard mixtures data showed that the joint-action models are better than the partial least-squares (PLS) model. Then, the Good2bad value, which could reflect components' importance based on Monte Carlo sampling, was coupled with the joint-action models for screening of active components. A compound (baohuoside I) and a component composed of compounds with retention times in the 6.9-7.9 min range were selected by our method. Their inhibition rates were higher than icariin, the key bioactive compound in Herba epimedii, which could inhibit osteoclast differentiation and bone resorption in a previous study. Meanwhile, the half-maximal effective concentration, namely, EC50 value of the selected component was 7.54 µg/mL, much smaller than that of baohuoside I-77 µg/mL-which indicated that there is synergistic action between compounds in the selected component. The results clearly show our proposed method is simple and effective in screening the most-bioactive components and compounds, as well as drug-lead components, from herbal medicines.

Two new compounds from Melodinus suaveolens.

Two new compounds, 19-hydroxy-melodinine K (1) and melodiside (2), and 25 known compounds were isolated from leaves and twigs of Melodinus suaveolens. Their structures were elucidated based on 1- and 2-D NMR, FTIR, UV and MS spectroscopic data. 19-hydroxy-melodinine K showed cytotoxic activity against MDA-MB-231 breast, BCG-823 gastric, SW480 colon and Hela cancer cells.

DanHong injection dose-dependently varies amino acid metabolites and metabolic pathways in the treatment of rats with cerebral ischemia.

AIM: To determine how the relative amino acid contents and metabolic pathways regulate the pharmacological phenotypes in rats with cerebral ischemia after treatment with varying doses of DanHong injection (DHI). METHODS: Adult male rats underwent middle cerebral artery occlusion (MCAO), and were injected with DHI (DH-1: 1 mL/kg; DH-2: 2.5 mL/kg; DH-3: 5 mL/kg, and DH-4: 10 mL/kg, iv) daily for 3 d. The neurological deficit score, body weights and infarct volume were assessed. Serum levels of 20 free amino acids were determined using HPLC, and the values were transformed through the quantitative analysis of the amino acids in the serum metabolic spectrum. Multivariate statistical analysis methods (PCA and PLS-DA) and web-based metabolomics tools (MetPa and MetaboAnalyst) were used to analyze the biological data sets for the amino acids. RESULTS: Administration of DHI dose-dependently decreased cerebral infarct volume, and ameliorated neurological deficits. A total of 5, 6, 7 and 7 non-overlapping metabolites were identified in the DH-1, DH-2, DH-3, and DH-4 groups, respectively. Eight metabolites were shared between the DHI groups and the vehicle group. In addition, the serum levels of glutamic acid, aspartic acid and serine increased with increasing DHI dose. A total of 3, 2, 2 and 5 non-overlapping metabolic pathways were identified in the DH-1, DH-2, DH-3 and DH-4 groups, respectively, and glycine, serine, threonine and histidine metabolism were identified as overlapping pathways among the 4 dose groups. CONCLUSION: Overlapping and non-overlapping amino acid metabolites and metabolic pathways are associated with the dose-dependent neuroprotective effect of DHI.

Synergistic inhibitory effect of Icariside II with Icaritin from Herba Epimedii on pre-osteoclastic RAW264.7 cell growth.

Increasing evidence shows the therapeutic superiority of herbal extracts in comparison to isolated single constituents. One of the reasons may be attributed to the synergy effect of compound combinations. Flavonoids from Herba Epimedii have been shown to have therapeutic effect against bone loss. Our previous study showed that Icariside II inhibited pre-osteoclast RAW264.7 growth. The aim of this study was to investigate whether the activity of Icariside II is synergized by other components of Herba Epimedii. The inhibitory activity of Icariside II was significantly enhanced in the presence of the extract of Herba Epimedii (EHE) at the ratio of 1:1, 1:5 and 1:10. Icaritin, another flavonoid constituent, was shown here to inhibit RAW264.7 growth in a dose-dependent manner. Further, we found that Icariside II, together with Icaritin, synergistically inhibited RAW264.7 growth. The synergistic effect is significant when the ratio of Icariside II and Icaritin was 10:1, 5:1, 1:1, 1:2, and 1:5, respectively. In conclusion, Icaritin were an active component. The inhibitory activity of Icariside II on pre-osteoclast RAW264.7 growth was synergized by Icaritin, which maybe contribute to the efficiency of Herba Epimedii extract on curing bone-related diseases, such as osteoporosis.

[Antitumor components screening of Stellera chamaejasme L. under the case of discrete distribution of active data].

This is to report the screening, extracting and validating antitumor components and compounds from Stellera chamaejasme L. under the case of discrete distribution of active data. In this work, different components from Stellera chamaejasme L. were collected by HPD macroporous resin and polyamide resin column, and their antitumor activity on A549 were tested by MTT assay. Activity results indicate that activity of components at 30-39 min is more potent than that of Stellera chamaejasme L. extract, and the activity of components at 33.97 min is equivalent to positive drug, cis-platinum at 100 microg x mL(-1), but with totally different mode of action. Under the case of discrete activity, the weight analysis is capable of screening active components and compounds from natural products.

[Antitumor effect of alcohol extracts from Stellera chamaejasme].

OBJECTIVE: To screen the best antitumor components of Stellera chamaejasme and their sensitive cell lines. METHOD: Sixteen different components of alcohol extracts from S. chamaejasme, including HH, H1-H8, JH and J1-J8, were got by gradient column chromatography eluted with alcohol in different concentrations. In the first screening, the solvent control group, the drug group, the positive group and the blank group were set up. Then the human cancer cell lines such as hepatocarcinoma BEL-7402, SK-HEP-1, and lung cancer A549, NCI-H157 were processed with the components, and the concentration for each drug group was 100 mg x L(-1). Thus, the 48 hour suppression ratio to the four kinds of cancer cells for each component were compared by the SRB method, to select the most inhibitive components and the most sensitive cell lines, which were used as the subjects of the second screening. In the second screening, each component including the concentration of 6.25, 12.5, 25, 50, 100 mg x L(-1) was used to treat the sensitive cell lines and the inhibition rates to each cell line of 24, 48, 72 h by the SRB assay were detected. Also, the IC50 of each component was calculated and their main chemical composition was analyzed by UPLC-MS. RESULT: The inhibition effect to the proliferation of the different cancer cells has great difference among 16 components, and the lung cancer cells are more sensitive to them than the hepatocarcinoma cells. Besides, the inhibition rates of JS, J6 and H8 are higher than the other components and their effect has a certain time and concentration dependence. At 72 h, the inhibition rate of each component ranges from (60.57 +/- 3.83)% to (96.66 +/- 0.51)% for lung cancer cells, and IC50 from (9.61 +/- 0.79) mg x L(-1) to (55.76 +/- 2.31) mg x L(-1). J5, J6 and H8 are the biflavonoids. CONCLUSION: The biflavonoids in alcohol extracts from S. chamaejasme have exerted a satisfactory inhibitory effect on the lung cancer cell proliferation.

[Screening of bioactive components in epimedium for osteoporosis treatment by model population analysis].

Epimedium was obtained from different habitats, and their bioactive components for inhibiting RAW264.7 were screened by MTT assay. Results indicate that epimedium from different habitats displayed significant different activities. By means of model population analysis (MPA), a latent bioactive component, baohuoside-I was got. Activity of baohuoside-I wasvalidated and prior to icariin. MPA can be used for bioactive components screening.

Taxol-induced alteration of intracellular amino-acid profile related to human cervical carcinoma HeLa cell death.

The anti-tumor action of Taxol was investigated in the changes of amino-acids involved in tumor cell survival. By tracing the intracellular amino-acid profiles of HeLa cells treated with non-conditioned and three conditioned media (Taxol, L-alanine, and Taxol + L-alanine), it was observed that an alteration of amino-acid metabolism participates in Taxol-induced death of HeLa cells. The contents of 18 out of 21 detected amino-acids are 5-95% and the ones of lysine and methionine are 158 and 117% of the corresponding contents in the control after treatment with Taxol for 24 h, respectively. Addition of L-alanine inhibited cell apoptosis upon Taxol treatment by partially blocking the increase of lysine and methionine and reversing decrease trend of alanine, glycine, and glutamic acid. These results suggest that interference of amino-acid metabolism might be an important mechanism of Taxol cytotoxicity.