Anticancer effects of crude extract from Melia toosendan Sieb. et Zucc on hepatocellular carcinoma in vitro and in vivo / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.</p><p><b>METHODS</b>Transonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.</p><p><b>RESULTS</b>HPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.</p><p><b>CONCLUSIONS</b>Toosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.</p>

Inheritance in endophytic fungi from Melia toosendan and diversity of genes of PKS and NRPS / 中草药

Objective: To explore the inheritance in endophytic fungi from Melia toosendan and the diversity of the genes of PKS and NRPS and to lay the foundation of looking for the synthesis of the potential strains of bioactive substances. Methods: To identify the morphology of 39 strains of endophytic fungi separated from the medicinal plant M. toosendan; The ITS sequences and the genes of PKS and NRPS were obtained using PCR method, compared by BLAST and analyzed by phylogenesis after sequencing. Results: The morphological identification and phylogenetic analysis on 39 strains showed that all strains of endophytic fungi from M. toosendan belonged to 25 categories, mainly Penicillium, Aspergillus, and Trichoderma. The dominant fungus species were Aspergillus (17.9%), and Penicillium (15.4%). PKS (12) and NRPS (6) genes were detected in this study, All NRPS genes were Penicillium. Conclusion: The endophytic fungi of M. toosendan have a rich genetic diversity and strong potential of synthesis of bioactive substances. Further research on Penicillium and Aspergillus should be carried out.

Chromatography fingerprint of the constituents of Melia toosendan Sieb / 中成药

AIM:To establish the chromatography fingerprint of the constituents of Melia toosendan Sieb from different areas in order to provide a base for the identification of its quality. METHODS: A gradient separated method was applied. Column:Inertsil ODS-3 C_ 18 ,mobile phase:acetonitrile-water,detection wavelength:270 nm,flow rate:1.0 mL/min,column temperature:25 ℃. RESULTS: To establish the chromatography fingerprint of the constituents of Melia toosendan Sieb., make the technical parameters for its quality controlling,and mark 39 main peaks as its characteristic fingerprint. CONCLUSION: The distribution of constituents of Melia toosendan Sieb differ a little from different areas, but the propotion of the constituents differ greatly, with the Melia toosendan Sieb.from the same area , the distribution and propotion differ a little. This method is reproducible, simple and easy, and can be use to provide a base for the quality control of Melia toosendan Sieb.