- 5-Methoxyleoligin, a Lignan from Edelweiss, Stimulates CYP26B1-Dependent Angiogenesis In Vitro and Induces Arteriogenesis in Infarcted Rat Hearts In Vivo

Figure S1. Overexpression of TXNIP does not interfere with 5-Methoxyleoligin mediated increase in endothelial tube formation. HUVECs stably expressing thioredoxin interacting protein (TXNIP) and controls were incubated with 5ML and subjected to capillary tube formation as outlined in the Material and Methods section. In contrast to CYP1A1 and CYP26B1 knock down, overexpression of TXNIP-which was found to be down-regulated by 5ML had no significant effect on tube spontaneous and 5ML induced formation. Figure S2. 5ML inhibits the proliferation of SMCs and increases the proliferation of HUVECs. To examine the potential effect of 5ML on the proliferation of HUVECs and vascular SMCs in vitro, we performed cellular metabolic assays (XTT) which allows conclusions about the proliferative activity of cells. XTT-based analysis revealed that incubation of HUVECs with 5ML (10 µM) activates significantly the proliferation of the cells, but lower concentrations of 1 µM had no effect. Contrasting results delivered the incubation of vascular smooth muscle cells with 5ML in vitro: concentrations of 1 µM 5ML had no effect on the proliferation. However, incubation with a higher concentration of 10 µM 5ML significantly inhibits the proliferation. Figure S3. 5ML is a stimulator of capillary tube formation with HMVECs. As with HUVECs, 5ML is also able to increase the tube formation with HMVECs (dose dependent increase). Figure S4. 5ML-treated infarction areas show a non-significant increase in the small arteries-density compared to the control hearts. As already mentioned, 5ML is able to significantly increase the number of arterioles in the infarction and peri-infarction area. Further histological analysis revealed there is a non-significant trend towards an increased number of small arteries in the infarction area.
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