| Objective Extraction and separation of the aerial parts of Bletilla striata, screening of the Antineoplastic effective part of inhibiting proliferation of tumor cells and studying the effect on this part to investigate the in vitro and vivo anti-tumor-related mechanism of action to provide a theoretical basis for the development and utilization of Bletilla striata.Methods (1) Using the ethanol Refluxing methods and ethanol precipitation to collect the extracts of the aerial parts of Bletilla striata and HPLC was used to analyze the chemical composition of extracts,then the antineoplastic effective part was selected. (2) Effects of the alcohol extraction from Bletilla striata aerial parts on the growth of the lung adenocarcinoma A549 cells were evaluated by MTS. (3) Analyzed the impact of the alcohol extraction from Bletilla striata aerial parts on the apoptosis of A549 by Hoechst-PI staining. (4) The influence of the alcohol extraction from Bletilla striata aerial parts on A549 cell migration was measured in two dimensions. (5) The expression of bcl-2, bax and ICAM-1 gene in A549 was evaluated by RT-PCR. (6) Hepatic cancer H22 model in mice was performed to study the anti-hepatoma effects of the alcohol extraction from Bletilla striata aerial parts in vivo; using the immunohistochemistry to analyze the expression of Bax and Survivin protein in tumor tissue and testing the expression of mRNA of Bcl-2 and Bax.Results (1)Compared with tubers of rhizome Bletillae striatae, the small molecule content higher than the content of polysaccharides in the aerial parts of Bletilla striata and the alcohol extraction from Bletilla striata aerial parts (small molecules) Mainly includes chemical compounds such as phenanthrenes, dihydro-phenanthrene. (2) The antineoplastic effective part in the extracts of the aerial parts of Bletilla striata is alcohol extract(the alcohol extraction from the Stem and leaf of Bletilla striata was mixed as effective parts of alcohol extract). (3) The lung adenocarcinoma A549 cells were treated with different concentrations of the alcohol extraction from Bletilla striata aerial parts separately for 24 h,48 h and 72 h, and we found the extraction can significantly reduce A549 survival rate in a dose-dependent manner. (4) Hoechst-PI double staining showed the following results:a large number of apoptotic cells was observed after Hoechst staining and cell morphology also changed (including the nucleus becomes smaller) when 40 μg/mL of the alcohol extraction from Bletilla striata aerial parts treated at A549, then a large number of cells died and the strong red fluorescence was observed after PI staining when 50 μg/mL of the extraction treated at A549. (5) Observed the healing assay, the alcohol extraction from Bletilla striata aerial parts can inhibit metastasis of lung adenocarcinoma cell on plane. (6) Promoting apoptosis of lung adenocarcinoma cells primarily by inhibiting the antiapoptotic gene of Bcl-2, promoting the expression of the promoting apoptosis gene of Bax to realize. It also can inhibit ICAM-lgene to reduce metastasis of lung adenocarcinoma cells. (7) The alcohol extraction from Bletilla striata aerial parts can significantly inhibit tumor growth in H22 mice; it’s also able to adjust the body’s physiological condition in mice, improve the quality of life and the body’s immune function, and has little toxic effect on the liver and the kidney toxicity. (8) The alcohol extraction from Bletilla striata aerial parts can increase the expression of Bax and decrease the expression of Survivin in tumor tissue of H22 mice, and a significant effect on moderate doses; It by inhibiting the antiapoptotic gene of Bcl-2, promoting the expression of the promoting apoptosis gene of Bax to induce apoptosis of tumor cell to realize anti-tumor.Conclusion The alcohol extraction from Bletilla striata aerial parts (small molecules) mainly includes chemical compounds such as phenanthrenes, dihydro-phenanthrene, which has a significant inhibitory effect on lung adenocarcinoma A549 cells, and there is a dose-response relationship.it primarily by inhibiting the antiapoptotic gene of Bcl-2, promoting the expression of the promoting apoptosis gene of Bax to realize. It also can inhibit ICAM-lgene to reduce metastasis of lung adenocarcinoma cells. In the meantime, the alcohol extraction from Bletilla striata aerial parts can adjust the body’s physiological condition in mice, improve the quality of life and the body’s immune function, increase the expression of Bax and decrease the expression of Survivin to induce apoptosis of tumor cell to realize anti-tumor. |
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