Article Figures & Data
Figures
- Figure 1.
Cell proliferation. Cell growth of MTC-SK cells was determined following treatment with different concentrations of isopteropodine (A1) and pteropodine (A2) from Uncaria tomentosa (UNC). 100 μM of A1 as well as 50-100 μM of A2 significantly inhibited cell proliferation of MTC-SK cells. Co represents untreated MTC-SK control cells. Error bars represent standard deviation σ.
- Figure 2.
Cell viability: A, MTC-SK cells were treated with different concentrations of the methanol extract (M) as well as the alkaloids isopteropodine (A1) and pteropodine (A2) from Uncaria tomentosa (UNC) and cell viability was measured. Treatment with 100-200 μM of the M fraction of UNC did not show any effect on cell viability of MTC-SK cells. In contrast, A1 and A2 of UNC (100 - 200 μM) significantly decreased the enzymatic activity of mitochondrial dehydrogenase of MTC-SK cells in the WST-1 assay. B, The same concentrations of M, A1 and A2 did not affect cell viability of normal human skin fibroblasts (HF-SAR cells) (positive control). Error bars represent s.e.m. All experiments were performed in triplicates.
- Figure 3.
DAPI-stain: A, Untreated control cells; B, MTC-SK cells were treated with A1 from Uncaria tomentosa (UNC), bright fluorescent chromatin indicates compact apoptotic cells.
- Figure 4.
DNA-Cycle: The sub-G1 (M4) ratio in MTC-SK cells treated with Uncaria tomentosa (UNC) was examined using flow cytometry. Cells were harvested after 12 h, 24 h and 48 h then they were fixed stained and analyzed for DNA content. The distribution and percentage of cells in sub-G1 phase (M4), G1 (M1), S (M2) and G2/M (M3) phase of the cell cycle are indicated. The sub-G1 peak displays cells with a lower DNA content. A) Treatment of MTC-SK cells with 200 μM M fraction from UNC did not affect the cell cycle after 12, 24 and 48 h. Furthermore, no apoptotic signs became visible. B) Following administration of 200 μM of A1 a sub-G1 peak was detectable in MTC-SK cells after 12 h, which was enhanced after 48 h. C) Cells were treated with 200 μM A2 of UNC, a sub-G1 peak was detectable in MTC-SK cells after 12 h and increased after 48 h.
- Figure 5.
Caspase-3 activity was examined following treatment with different concentrations of the alkaloid-poor methanol fraction (M), as well as the alkaloids isopteropodine (A1) and pteropodine (A2) from Uncaria tomentosa (UNC). A, MTC-SK cells treated with 100-200 μM of the M fraction of UNC did not show cleaved caspase-3 increases. In contrast, a significant increase in caspase-3 activity was observed in cells treated with 100-200 μM of A1 of UNC. Cells treated with A2 (100-200 μM) displayed less cleaved caspase-3 increase than cells treated with A1. B, Neither treatment with the M fraction nor with A1 and A2 (150-200 μM) affected caspase-3 activity of HF-SAR cells.
- Figure 6.
Cytometric Bead Array measurement: The simultaneous measurement of cleaved caspase-3, Bcl-2 and PARP was performed with the CBA kit. Following treatment of MTC-SK cells with 200 μM A1 (A) and A2 (B) of UNC, an increase of cleaved caspase-3 as well as an enhancement of PARP was observed. However, in MTC-SK cells treated with A1, the highest cleaved caspase-3 amount was detected after 24 h, whereas a high amount of cleaved caspase-3 in cells treated with A2 was already detected after 8 h. No significant change in Bcl-2 expression was detected. Normal fibroblasts showed no cleaved caspase-3, no enhancement of PARP and no change in Bcl-2 expression (data not shown).
