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Objective To optimize the extraction process of mixed volatile oil from Forsythiae Fructus, Saposhnikoviae Radix and Magnoliae Flos and inclusion process of β-cyclodextrin (β-CD). Methods With yield ratio of volatile oil as evaluation index, single factor experiments were used to study the extraction process of volatile oil;saturated aqueous solution was used, with inclusion rate of volatile oil as index, and orthogonal design was adopted to examine effects of charge ratio of volatile oil and β-CD, inclusion temperature and inclusion time on the inclusion process; X-ray scattering technique, infrared spectroscopy and scanning electron microscopy were used to characterize the inclusion compound. Results The optimum extraction process of volatile oil was soaking fine powder extracted 5 hours with 10 folds the amount of water. The optimum conditions of inclusion process were as follows:mixed ratio of volatile oil (mL) and β-CD (g) was 1:10; inclusion temperature was 50 ℃; the inclusion time was 2 h. X-ray scattering technique, infrared spectroscopy and scanning electron microscopy proved the inclusion compound had been formed. Conclusion Optimum extraction and inclusion processes are stable and feasible, and can provide research foundation for further research and development of preparation.
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Tumor neovascularization and vasculogenic mimicry are two important angiogenesis pathways, which play an important role in the occurrence, development and metastasis of breast cancer. TCM has confirmed efficacy in the prevention and treatment of breast cancer. In recent years, research on inhibition of tumor angiogenesis is also gradually in-depth. This article reviewed the progress of TCM in the dual pathways of breast cancer angiogenesis and its mechanism, and provided references for clinical medication and research and development of new medicine.
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Objective To observe the effects of tetra-atsenic oxide (As4O6) on the proliferation, migration and invasion of human breast cancer MCF-7 cells; To explore its potential mechanism. Methods The human breast cancer MCF-7 cells were regarded as the research object and cultured in vitro, with different concentrations of As4O6 using to intervene in MCF-7 cells. The cell proliferation was detected by MTT assay; the flow cytometry and wound-healing assay were used to detect the cell apoptosis and migration, respectively. The expressions of Cyclin E1, Cyclin A2, Caspase 3, p21 and MMP-9 mRNA were accessed by semi quantitative RT-PCR. Results As4O6 had a significant inhibitory effect on the proliferation of MCF-7 cells in a dose dependent manner. Compared with the control group (0 μmol/L), the apoptosis rate increased significantly when the concentration of As4O6 was 9, 12, 15 μmol/L (P<0.01). Either As4O6 at high (3 μmol/L) or low (1 μmol/L) concentration could effectively inhibit the migration of MCF-7 cells (P<0.01). With the increasing concentration of As4O6, the expressions of Cyclin E1, Caspase 3, and p21 mRNA significantly increased, while the expressions of Cyclin A2 and MMP-9 mRNA significantly decreased (P<0.05, P<0.01). Conclusion As4O6 can significantly inhibit the proliferation, cycle, invasion and migration of breast cancer MCF-7 cells, and the mechanism may be related to the increase of expressions of cyclin E1, caspase 3, p21 and inhibition of expressions of cyclin A2 and MMP-9.
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Objective To observe toxic symptoms and signs , toxic damage extents and reversibility in rats after oral administration of Tangwang Mingmu granules .Methods Four dose groups with 40 rats in each group were designed in this study, including control group fed with distilled water and three groups at different dosages of the test drug .Tangwang Mingmu granules were orally administered to SD rats at the dosage of 8.4, 4.2 and 2.1 g/kg for 3 weeks and 14.0, 8.4 and 4.2 g/kg for 23 weeks, for 26 consecutive weeks .The general state of the rats was observed every day , while body mass and food consumption were calculated once a week .Halfway through and at the end of the administration (13 and 26 weeks) and after four weeks of recovery, parameters of body mass, hematology, hematological biochemistry, organ/body mass ratio and histopathology were measured .Results Compared with the control group at the same time-point, body mass of male rats in the other three groups was slightly reduced .Food consumption in high and medium dose groups was reduced (P<0.05), MCHC, ALT, TBIL and Na +in high dose group were decreased (P<0.05), TP, ALB and D-BIL were increased (P<0.05), the mean body mass and relative organ weight of thymus in medium dose male rats were decreased (P<0.05), relative organ weight of the liver and kidney in high dose male rats was increased (P<0.05), and focal chronic inflammation to different extent was observed in the liver , kidney and prostate gland .No dose-effect relationship was found in these perturbations that were all within the normal range of animals .No significant drug-related pathological changes were found.Conclusion The NOAEL of Tangwang Mingmu granules is considered to be 14.0 g/kg body mass/day (equal to 50 times the proposed clinical adult dosage ) for the 26-week repeated dose oral toxicity study in male andfemale rats.
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Objective To observe the effects ofTrametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on the migration and invasion of human breast cancer MCF-7 cells and relevant factors; To discuss relevant mechanism of action.Methods Breast carcinoma cell line MCF-7 was used as research subject in this experiment. Control group, Isatidis Radix group,Trametes robiniophila Murr group, andTrametes robiniophila Murr and Isatidis Radix group were included in the experiment. The effects ofTrametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on MCF-7 cell proliferation were measured by MTT method. Cell scratch assay, transwell assay and adhesion assay were used to measure the effects ofTrametes robiniophila Murrand Isatidis Radix bidirectional fermentation products on the migration, invasion and adhesion capability of MCF-7 cells, respectively. The effects ofTrametes robiniophila Murrand Isatidis Radix bidirectional fermentation products on the mRNA expression of MMP-9 and Vimentin were measured by RT-PCR.Results Compared with Isatidis Radix group andTrametes robiniophila Murrgroup, Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products could significantly inhibit the proliferation, migration, invasion and adhesion capability of MCF-7 cells (P<0.05). Similarly,Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products reduced the mRNA expression of MMP-9 and Vimentin (P<0.05).ConclusionTrametes robiniophila Murrand Isatidis Radix bidirectional fermentation products may down-regulate the expression of MMP-9 and Vimentin to inhibit the migration, invasion and adhesion capabilities of MCF-7 cells.
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Objective To observe the mechanism of action of different extracts ofTangwang Mingmu Granules on high glucose induced VEGF and IL-1α gene and protein expressions in vascular endothelial cells.Methods Human umbilical vein endothelial cells EA.hy926 were divided into six groups: blank, high glucose,Tangwang Mingmu Granules, extract 1 (glycoside and flavonoid), extract 2 (organic acid and polysaccharides) and extract 3 (alkaloids) groups. High concentration glucose was used to establish the high glucose model in EA.hy926 cells. The expressions of VEGF and IL-1α mRNA were detected by semi-quantitative RT-PCR. The contents of VEGF and IL-1α protein were tested by ELISA.Results The gene expression and protein levels of VEGF and IL-1α were significantly up-regulated under the high glucose condition (P extract 3> extract 2.Conclusion The action intensity of glycosides and flavonoids, alkaloids, organic acids and polysaccharides on VEGF and IL-1α expression inTangwang Mingmu Granules weakens in sequence.
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ObjectiveTo observe the effect of different extracts ofTangwang Mingmu Granule on hypoxia induced gene expressions in vascular endothelial cells.Methods COCl2 intervention cells were used to copy hypoxia models. Human umbilical vein endothelial cells EA. Hy926 were divided into blank group, hypoxia model group,Tangwang Mingmu Granule group, extract 1 (glycosides and flavonoids) group, extract 2 (orgain acids and polysaccharides) group and extract 3 (alkaloids) group. The changes in gene expressions of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α mRNA were detected by semi-quantitative RT-PCR.ResultsThe gene expression levels of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α were significantly up-regulated under the hypoxic condition (Palkaloids>organic acids and polysaccharides inTangwang Mingmu Granule.
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Objective To observe the preventive and therapeutic effects of the new process extracts ofTangwang Mingmu Granule on rats with diabetic retinopathy.Methods The best preparation process technology of drug extraction was improved and selected based on the traditional water decoction of Chinese medicine. The active ingredient of two processes were detected and compared by HPLC. In vitro and in vivo pharmacodynamic evaluation models were used to evaluate the drug effects and analyze its mechanism. MTT assay was used to analyze the inhibitory effects of extracts in the two processes on EA.hy 926 cell proliferation in vitro. The effects of two extracts on STZ induced rat diabetic microvascular disease and its related factors were observed by retinal slice staining and ELISA.Results Compared with the traditional process, the active ingredients of the extracts were improved by the new technology ofTangwang Mingmu Granule. The preparation process ofTangwang Mingmu Granule could significantly improve the diabetic rat retinal microvascular pathological state, and reduce the protein expression levels of angiogenesis related factors.Conclusion The new preparation process could effectively improve the active ingredients in the herbal extract. The extract of new technology showed a significantly effect on the diabetic rats in retinal microvascular pathology state. The mechanism may be related to the regulation of vascular cell adhesion molecules and inflammatory factors.
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Allergic rhinitis (AR) is one kind of nose mucous membrane inflammation reactiveness disease, which is mediated by IgE for the environment sensitizin. The major clinical symptoms are: nose itch, the sneeze, nasal discharge, nasal obstruction and so on. It is thought that the acient disease named "Bi Qiu" has included AR. By comparing the AR clinical manifestation characteristic with trditional Chinese medicine (TCM) "Feng/wind" induced disease characteristic and the "Feng/wind" disease clinical manifestation characteristic, we can know that AR should belong to the "Feng/wind" disease category of TCM. It is shown that the "Feng/ wind" medicine is the most windly appeared in AR treatment formulas of TCM clinical research data (in each formular) in the past 10 years. To aim directly at three knids of problem in TCM clinical research, we discussed five problems which should pay close attention to the feature research, therefor to provide a mirror for AR clinical and experimental studies by TCM.