Astragalus extract inhibits destruction of gastric cancer cells to mesothelial cells by anti-apoptosis

Figure 1 Morphological changes in human peritoneal mesothelial cells under phase contrast microscopy (× 40). A: Morphology of mesothelial cells cultured in serum free DMEM; B: Exfoliation and naked areas of mesothelial cells after treatment with Astragalus injection; C: Morphological changes in mesothelial cells after treatment with 200 &mgr;g/mL Astragalus injection; D: No typical morphological changes in mesothelial cells after treatment with DMEM + 200 &mgr;g/mL Astragalus injection.

Figure 2 Mesothelial cells under electron microscope after incubation with and without SF-CM. A: Normal nuclei and endoplasmic reticulum of control cells; B: Condensation of nuclear chromatin (arrow in Figure 2B); C: Wrinkling of nuclear membrane of cells after treatment with Astragalus injection (arrow in Figure 2C); D: Dilated endoplasmic reticulum (arrow in Figure 2D).

Figure 3 Viability of mesothelial cells after treatment with SF-CM and Astragalus injection. Mesothelial cells (5 × 10³ cells/well) were treated with SF-CM, 200 &mgr;g Astragalus injection, and SF-CM plus 200 &mgr;g Astragalus injection for 12, 24 and 48 h, respectively. Mesothelial cells cultured in serum free DMEM served as controls. MTT assays were performed in triplicate for each data point.

Figure 4 Effect of MKN45 and/or Astragalus injection on cell-cycle distribution in mesothelial cells treated for 12 h. DMEM with an apoptotic rate of 0.52% (A), SF-CM with an apoptotic rate of 10.03% (B), SF-CM plus 200 &mgr;g Astragalus injection with an apoptotic rate of 1.57% (C), DMEM + 200 μg/mL Astragalus injection with an apoptotic rate of 0.25% (D). Samples were analyzed by flow cytometry as described in “Materials and methods” section. Apoptotic peak is shown in green.

Figure 5 Percentages of mesothelial cells in sub-G1 group (apoptosis) after treatment with control, MKN45, MKN45 + 200 &mgr;g/mL Astragalus injection and 200 &mgr;g/mL Astragalus injection for various periods of time. ^bP < 0.01 vs 12 and 24 h.

Figure 6 Apoptosis of mesothelial cells treated for 48 h. A: serum free DMEM; B: MKN45; C: MKN45 + 200 &mgr;g/mL Astragalus injection; D: DMEM + 200 &mgr;g/mL Astragalus injection; E: GES-1 with AO/EB staining. Cells containing normal nuclear chromatin exhibit green nuclear staining. Cells containing fragmented nuclear chromatin exhibit orange to red nuclear staining.

Figure 7 Western blotting analyses of Bcl-2 and Bax proteins. Mesothelial cells were treated for 24 h with serum free DMEM, MKN45, MKN45 + 200 &mgr;g/mL Astragalus injection (MKN45 + A), DMEM + 200 &mgr;g/mL Astragalus injection, respectively.