Study On The Mechanisms And Anti- Tumor Active Of Extracts From Ampelopsis Megalophylla

Ampelopsis megalophylla Diels et.Gilg is a medicinal plant of the genus Ampelopsis, its medicinal part is tender leaves and stems. It could be called “Meicha” because there will be white rime on the surface after its leaves and stems scalded slightly in hot boiling water and dried at ventilated place. It is neutral in nature, sour and bitter tasting, with the functions of the activating blood, resolving stasis, stopping bleeding, clearing heat and toxins. Based on the previous research results, anti-tumor active ingredients of A. Megalophylla and its mechanism were studied in this paper.In preliminary research, our team has been completed extracted and isolated with the solvent method, and got ethanol extracts, the petroleum ether extracts, Eto Ac extracts and water extracts. Total flavones extracts and water detection extracts were obtained through percolation and water detection methods, respectively. The three kinds of tumor cells, Hela, SMMC-7721 and A549 cells were selected to screen anti-tumor activities for the extracts of A. Megalophylla. The experimental results indicated that the ethanol, Eto Ac and total flavones extracts were the main active fractions for the inhibition of Hela cells proliferation; the Eto Ac and total flavones extracts were the main active fractions for the inhibition of SMMC-7721 cells proliferation; the ethanol and Eto Ac extracts were the main active fractions for the inhibition of A549 cells proliferation. Hela and SMMC-7721 cells were selected to screen anti-tumor activity for twelve compounds from A. Megalophylla. The results showed that four compounds, Ampelospin, Myricetin, Gallic acid and Emodin, could strongly inhibit the proliferation of Hela cells. Emodin could strongly inhibit the proliferation of SMMC-7721 cells.Take Hela cells as subject, we studied anti-tumor mechanism of Ampelospin(AMP) through using Hochest33258 staining, propidium iodide(PI) staining, Annexin V-FITC/PI double staining, rhodamine123(Rh-123) and western blot analysis. The results suggested that AMP could inhibit the proliferation of Hela cells and induce apoptosis, apoptotic bodies could be found under fluorescent microscopy; Flow cytometry method showed AMP could make Hela cells cycle arrested in S phase. Apoptotic percent presented a dose-dependent in a certain range, meanwhile, mitochondrial membrane potential was markedly decreased. The expression of Caspase-3, Caspase-9 and Cytc proteins dramatically increased after incubation with AMP at concentrations of 0, 30, 40, 50, 60 and 70 μM for 12 h. Furthermore, different doses of AMP on Hela cells for 12 h resulted in decreasing of bcl-2 protein levels and increasing of bax protein expression, leading to a decrease of bcl-2/bax ratio. Through analyzing the experimental results, AMP may induced apoptosis in Hela cells via mitochondrial pathway.