ABSTRACT
Juvenile zebrafish were used to screen the active components of Lycii Fructus for improving osteoporosis. The screening results were further verified by zebrafish adult osteoporosis model and the action mechanism was explored. Prednisolone was used as the inducer to build osteoporosis models of juvenile and adult zebrafish, and 9 groups of samples of different extracts and chemical parts of Lycii Fructus were given. Alizarin red staining was applied for observing the scale matrix mineralization and bone resorption. The activities of osteoblasts and osteoclasts were detected using alkaline phosphatase (ALP) and tartrate resistant acid phosphatase (TRAP/TRACP) staining. The expressions of bone metabolism-related genes alp, osteoprotectin (opn), osteoblast specific transcription factor (sp7), cathepsin K (ctsk), tracp, and Runt family transcription factor 2b (runx2b) in each group were determined using quantitative polymerase chain reaction. The results showed that all components of Lycii Fructus improved the formation area of the first vertebrae, the staining light density value, and the number of vertebrae joints in juvenile zebrafish and the Lycium barbarum polysaccharide (LBP) treatment group exerted the best effect. In addition, LBP prevented the formation of bone resorption lacunae in zebrafish scales, increased ALP activity, decreased TRAP activity, up-regulated the alp, sp7, and opn genes, and lowered the expressions of ctsk and tracp genes. In conclusion, LBP regulated the activity of osteoblasts and osteoclasts, reduced bone resorption, promoted bone formation and enhanced bone density, which might be the main anti-osteoporosis active fraction of Lycii Fructus. This study provided modern scientific evidence for the scientific connotation of the traditional effect of "strengthening bones and muscles" of Lycii Fructus, provided the reference for the evaluation of the anti-osteoporosis activity of traditional Chinese medicine based on zebrafish adult model, and provided beneficial enlightenment for the bone health needs of the aging society population.
ABSTRACT
Platelet adhesion is a key process in thrombosis. Anti-platelet adhesion effect of some Chinese medicines for promoting blood circulation and removing blood stasis (PBCRBS) has been reported, but their relative efficacies as a whole and specific targets remained unclear. This paper combined activity screening, drug compatibility analysis, pathway clustering, target prediction, and molecular docking to explore the mechanism of anti-platelet adhesion by PBCRBS Chinese medicine. Screening the activity of anti-platelet adhesion of 58 commercially available PBCRBS Chinese patent medicines showed that about 50.0% significantly inhibit ADP-induced platelet adhesion in vitro, and about 96.6% significantly inhibit thrombin-induced platelet adhesion in vitro. The animal experiment involved was approved by the Animal Ethics Committee of Tianjin International Biomedical Research Institute. Combined with the auxiliary platform for TCM (V2.0) inheritance showed that the compatibility of Danshen-Chuanxiong was used most frequently among the top 20 active proprietary Chinese patent medicines. IPA network analysis revealed that IL-1, APP and CCL2 might be the key targets for anti-platelet adhesion function of Danshen-Chuanxiong against atherosclerosis, neuroinflammation and chemokine signaling pathways as the main mechanisms. Molecular docking analysis confirmed the interaction between one of the active compounds shared by Danshen and Chuanxiong, i.e. chlorogenic acid, with its target CCL2. This study provides TCM theory guidance and experimental support for targeting platelet adhesion in anti-thrombosis therapy by Chinese medicine for promoting blood circulation and removing blood stasis.
ABSTRACT
Objective To establish an effective method to rapidly detect acetylcholinesterase inhibitors from natural products, and provide support for the treatment of Alzheimer’s disease with Chinese medicine and its active ingredients. Methods The activity of restraining acetylcholinesterase of crude extracts of a variety of medicinal plants were evaluated by applying Ellman’s colorimetric, and the better activity of the extract sample solution was filtered out. The active ingredients of medicinal plants were identified and separated by the application of AS/UF-UPLC-MS. Results According to the experimental data, Corydais yanhusuo, Phellodendron chinense, Uncaria rhynchophylla, and Gynura segetum had significant restraining acetylcholinesterase activity. Rapid screening showed that 3α-dihydrocadambine, 3α-isodihydrocadambine, rhyncholphylline, isorhynchophylline, corynoxine, and isocorynoxine were active ingredients in U. rhynchophylla. Conclusion The results suggest that it is feasible to screen acetylcholinesterase inhibitors from natural organism plants. Using AS/UF-UPLC-MS methods, leading compounds for the development of new drugs for Alzheimer’s disease would be developed and provided.
ABSTRACT
Objective To investigate the applicability of zebra fish thrombosis model in antithrombotic activity screening of Chinese materia medica.Methods The living zebra fish thrombosis model was induced by adrenaline hydrochloride. Zebra fish were randomly divided into blank control group, model group, positive medicine group and medication group. Each group was given the corresponding medicine or embryo culture water. O-anisidine staining solution was used to stain and calculate the staining intensity of erythrocytes in zebra fish heart, and quantitative analysis was carried out. The platelet aggregation of transgenic zebra fish was observed and under qualitative analysis. Results Compared with the model group, 100μg/mL salvianolic acid B, 300, 900μg/mL aqueous extract of Salvia miltiorrhiza, 45μg/mL 95% ethanol extract and 400, 1200μg/mL hypothalamus could significantly inhibited the formation of zebra fish thrombosis (P<0.01).ConclusionZebra fish thrombosis model has good applicability in antithrombotic activity screening of Chinese materia medica.
ABSTRACT
The increasingly apparent liver injury problems of bone strengthening Chinese medicines have brought challenges for clinical application, and it is necessary to consider both effectiveness and safety in screening anti-osteoporosis Chinese medicines. Metabolic transformation is closely related to drug efficacy and toxicity, so it is significant to comprehensively consider metabolism-action/toxicity(M-Act/Tox) for screening anti-osteoporosis Chinese medicines. The current evaluation models and the number of compounds(including metabolites) severely restrict efficient screening in vivo. By referring to previous relevant research and domestic and abroad literature, zebrafish M-Act/Tox integrative method was put forward for efficiently screening anti-osteoporosis herb medicines, which has organically integrated zebrafish metabolism model, osteoporosis model and toxicity evaluation method. This method can break through the bottleneck and blind spots that trace compositions can't achieve efficient and integrated in vivo evaluation, and realize both efficient and comprehensive screening on anti-osteoporosis traditional medicines based on in vivo process taking both safety and effectiveness into account, which is significant to accelerate discovery of effective and safe innovative traditional Chinese medicines for osteoporosis.
ABSTRACT
Objective: To screen the active compounds in Sini Decoction showing the potential to inhibit tumor necrosis factor α (TNFα) to alleviate Doxorubicin (DOX)-induced heart failure. Methods: A chemical database of Sini Decoction was constructed from literature research. The generated pharmacophore models based on TNF-α used to screen active ingredients of Sini Decoction in the database by Discovery Studio 2.5. Molecular docking by Autodock 4.2 was adopted to demonstrate the hit compounds' affinities with TNFα. Furthermore, DOX-induced heart failure model on H9c2 cell line was constructed and cell viability was detected by CCK-8 to validate the therapeutic effect of potential active compounds. Results: The higenamine showed potential cardiovascular protective effect through virtual screening. And the activity was identified in vitro. Conclusion: In this study, we found that higenamine may inhibit TNF-α through virtual docking and validated that higenamine may have the potential of treatment for heart failure in the model of doxorubicin-induced myocardial toxicity to H9c2 cells.
ABSTRACT
OBJECTIVE: To study the chemical constituents of the extract of Arnebia euchroma (Royle) Johnst. and screen natural protein tyrosine phosphatase 1B (PTP1B) inhibitors. METHODS: Silica gel, MCI gel, ODS gel, and Sephadex LH-20 chromatographic techniques were used to study the chemical constituents of A. euchroma, the chemical structures were elucidated by analysis of physico-chemical and spectral data, and the inhibitory activity on PTP1B enzyme was tested in vitro. RESULTS: Eight compounds were obtained and their structures were identified as deoxyshikonin (1), shikonin (2), acetylshikonin (3), β, β'-dimethylacrylalkannin (4), quercetin (5), kaempferol (6), kaempferide (7), and β-sitosterol (8). Compounds 1-4 exhibited inhibitory activities on PTP1B with IC50 values of (0.80±0.16), (4.42±0.37), (1.02±0.13), and (0.36±0.08) μmol·L-1, respectively. The study of structure-activity relationship showed that the ring of naphthoquinone might be the key skeleton structure for the inhibition activity on PTP1B, and the 2-substituted long lipo-chain of naphthoquinone significantly affected the activity of these compounds: the increase in the polarity of the lipo-chain led to the reduction of activity, while the increase in the terminal double bond of the lipo-chain led to the enhancement of activity. CONCLUSION: Shikonin derivatives with 1, 4-naphthoquinone skeleton is a new type of leading compounds for the treatment of diabetes.
ABSTRACT
Objective: In this study, ultrafiltration membrane process is employed first to separate the polysaccharides from industrialization waste in Mailuoning Injection and to promote the resources utilization of the waste. Methods: The waste ethanol sediments in the production of Mailuoning Injection mainly include dendrobium polysaccharides, Achyranthes bidentata polysaccharides (ABPS), and honeysuckle polysaccharides, and the active components of polysaccharide with high content were selected through the immune activity. Six different relative molecular weight cut-off membranes ( 3 × 105) made of polyether sulfone (PES) were used. Six fractions were obtained and named as MFP1, MFP2, MFP3, MFP4, MFP5, and MFP6. Meanwhile, polysaccharides and protein content were determined by phenol-sulfuric acid colorimetry and Coomassie brilliant blue G-250 staining method; Polysaccharides species was analyzed by ELSD-HPLC; Immunocompetence of different fractions were investigated in vitro, including mouse spleen lymphocytes proliferation, transformation, and production of nitric oxide after acting on the macrophage RAW264.7. Results: The results showed MFP4 (3 × 104-1 × 105) have the highest content and immunocompetent, MFP5 (1 × 105-3 × 105) immunocompetent followed, compared to others, purity, and recovery of MFP4 are 76.80% and 10.81%, purity and recovery of MFP5 are 73.23% and 14.73%, respectively. Conclusion: The findings reveal MFP4 and MFP5 serves as our object of exploitation in the future; Moreover, separation of polysaccharides from "Mailuoning Injection" by the membrane is feasible with high recovery and low cost in the realization of circular economy.
ABSTRACT
We isolated 61 endophyte isolates from the bark of 4 plants, Ginkgo biloba L, Albizzia julibrissin Durazz, Ailanthus sltissima (Mill) Swingle and Melia azedarach L. At the test concentration of 200 ?g/mL, higher than 50% of antitumor activities were demonstrated with crude extracts from 45.9% of fungal culture in MTT assay. Six isolates, YX5, YX17, YX36, KL1, CC1 and CC5, still showed higher cytotoxicity at 50 ?g/mL. No isolates from A. julibrissin had inhibitory effect towards EC109 at the test concentration of 50 ?g/mL; while about 15.8% of isolates from G. biloba were active. IC50 of the extract from the most active isolate YX5 against EC109, HONE1 and HeLa were 18.3 ?g/mL, 3.6 ?g/mL and 6.5 ?g/mL, respectively. Our results indicate that endophytes from G. biloba could be regarded as a potent source of antitumor drugs.