Evaluation of angiogenic and neuroprotective potential of different extracts from three Veronica species

Figures & data

Table 1. Total phenolics, iridoids and phenylpropanoid glycosides in Veronica species.

Sample	Total phenolics (mg GAE/g DW)	Total iridoids (mg aucubin/g DW)	Total phenylpropanoids (mg acteoside/g DW)
V. jacquinii ME	195.4 ± 1.5	167.8 ± 3.2	18.7 ± 0.3
V. jacquinii AW	201.4 ± 3.1	249.6 ± 3.6	31.5 ± 0.4
V. teucrium ME	156.7 ± 0.9	231.0 ± 5.1	26.3 ± 0.5
V. teucrium AW	172.1 ± 1.5	322.9 ± 6.1	38.0 ± 0.6
V. urticifolia ME	167.6 ± 1.3	270.2 ± 4.5	13.4 ± 0.2
V. urticifolia AW	171.5 ± 2.2	359.7 ± 4.8	35.0 ± 0.6

Note: Values are means ± SD of three measurements.

ME = methanol; AW = acetone–water; GAE = gallic acid equivalents; DW = dry weight.

Table 2. Acteoside and aucubin contents in Veronica species.

Sample	Acteoside (mg/g DW)	Aucubin (mg/g DW)
V. jacquinii ME	38.02 ± 3.75	n.d.
V. jacquinii AW	29.28 ± 1.60	n.d.
V. teucrium ME	34.44 ± 1.07	n.d.
V. teucrium AW	32.89 ± 1.42	n.d.
V. urticifolia ME	108.74 ± 4.58	21.27 ± 1.13
V. urticifolia AW	88.37 ± 4.58	11.04 ± 0.33

Note: Values are means ± SD of three measurements.

ME = methanol; AW = acetone–water; DW = dry weight; n.d. = not determined.

Table 3. Influence of Veronica extracts on the viability of SH-SY5Y cells stressed with sodium nitroprusside (SNP) and hydrogen peroxide (H₂O₂).

	V. urticifolia		V. jacquinii		V. teucrium
Extract	AW	ME	AW	ME	AW	ME
Increase in cell viability (%)
SNP	11.0 ± 3.5	12.0 ± 3.6	12.0 ± 2.6	12.3 ± 2.1	9.7 ± 3.8	10.7 ± 2.5
H2O2	17.0 ± 3.6	13.0 ± 3.5	17.0 ± 4.6	19.0 ± 5.6	18.3 ± 4.9	21.3 ± 6.8

Note: Tested extracts were added as pretreatment 30 min before the addition of SNP/H₂O₂. The increase in cell viability is presented as a percentage of the basal level of viability (100%) observed in non-treated cells. Presented values are mean ± SEM obtained from at least three experiments. ME = methanol; AW = acetone–water.

Figure 1. Influence of Veronica extracts on the amount of thiobarbituric acid reactive species (TBARS) in the cells stressed with sodium nitroprusside (SNP)/hydrogen peroxide (H₂O₂). Cells were pretreated with 50 µg/ml of extracts 30 min before addition of the stressor: (a) 1 mM SNP or (b) 100 µM H₂O₂. The index of lipid peroxidation (ILP) was measured 24 h after the treatment. The results are presented as a percentage of the amount of TBARS produced in cells treated only with stressors, which represented 100%. Presented values are mean ± SEM obtained from at least three experiments. VU = Veronica urticifolia; VJ = Veronica jacquinii; VT = Veronica teucrium; AW = acetone–water; ME = methanol.

Figure 2. Influence of Veronica extracts on the amount of released superoxide () radicals in the cells stressed with sodium nitroprusside (SNP)/hydrogen peroxide (H₂O₂). Cells were pretreated with 50 µg/ml of extracts 30 min before addition of the stressor: (a) 1 mM SNP (A) or (b) 100 µM H₂O₂. The nitroblue tetrazolium (NBT) test was performed 24 h after the treatment. The results are presented as a percentage of the amount of radicals released by cells treated only with stressors, which represented 100%. Presented values are mean ± SEM obtained from at least three experiments. VU = Veronica urticifolia; VJ = Veronica jacquinii; VT = Veronica teucrium; AW = acetone–water; ME = methanol.

Table 4. Influence of Veronica extracts on endothelial cell adhesion.

		V. urticifolia		V. jacquinii		V. teucrium
Extract	Control	AW	ME	AW	ME	AW	ME
Influence on endothelial cell adhesion (%)	100.0 ± 2.1	108.3 ± 5.7	103.4 ± 19.2	110.4 ± 11.2	108.0 ± 16.1	111.0 ± 12.0	110.8 ± 7.4

Note: The EA.hy926 cells were pretreated with tested extracts (25 µg/ml) for 24 h and seeded on Matrigel-coated plates. Following 15 min incubation, the number of adhered cells was estimated using crystal violet staining. The results are presented as a percentage of cell adhesion, where the adhesive capacity of non-treated control cells was set at 100%.

Presented values are mean ± SD obtained from at least three experiments.

ME = methanol; AW = acetone–water.

Figure 3. Influence of Veronica extracts on the migratory capacity of EA.hy926 cells. Confluent monolayers of EA.hy926 cells were wounded and immediately after formation of scratches treated with investigated extracts (25 µg/ml). The healing process was monitored at indicated time-points and data are expressed as a percentage of scratch width covered by proliferating and/or migrating cells, where the healing capacity of the untreated control cells after 24 h was set at 100%. VU = Veronica urticifolia; VJ = Veronica jacquinii; VT = Veronica teucrium; AW = acetone–water; ME = methanol.

Figure 4. Influence of Veronica extracts on tube formation. The cells were seeded on Matrigel and treated with the investigated extracts (25 µg/ml). After 12 h of incubation, (a) photographs were taken, and (b) the number of formed loops was counted and compared. VU = Veronica urticifolia; VJ = Veronica jacquinii; VT = Veronica teucrium; AW = acetone–water; ME = methanol.