Exploration of collagenase, cyclooxigenases, angiogenesis and free radical processes as the putative pharmacological targets of Arum maculatum L.

Figures & data

Table 1. GC-MS data of the volatiles of the extracts of Arum maculatum roots.

No.	Compound	R1(min)	Formula	Exact Mass:	BP	% area
						30% EtOH extract	70% EtOH extract
1.	Benzamide, N-(2′-ethylphenyl)-(N-(2-Ethylphenyl) benzamide)	6.39	C15H15NO	225.115364	105	4.44	2.21
2.	n-Nonadecanol-1	6.71	C19H40O	284.307917	71	2.24	3.75
3.	2-Undecanethiol, 2-methyl-	6.99	C12H26S	202.175522	69	12.14	22.71
4.	Benzene, 1-ethyl-3-methyl (3-Ethyltoluene)	7.57	C9H12	120.0939	105	3.61	7.2
5.	Benzene, 1-ethyl-3-methyl	8.11	C9H12	120.0939	105	3.36	6.7
6.	3-Decyn-2-ol	8.35	C10H18O	154.135765	97	8.21	14.2
7.	Mesitylene (Benzene, 1,3,5-trimethyl)	8.62	C9H12	120.0939	105	13.94	19.22
8.	2-Hexyl-1-octanol	8.85	C14H30O	214.229666	57	8.8	12.4
9.	α-Pinene oxide (3-Oxatricyclo[4.1.1.0(2,4)]octane, 2,7,7-trimethyl)	9.87	C10H16O	152.120115	67	2.07	3.75
10.	Carveol	10.58	C10H16O	152.120115	83	4.89	5.2
11.	Benzene, (1-methylpropyl) (2-Phenylbutane)	10.81	C10H14	134.10955	105	2.66	3.92
12.	p-Cymene	11.19	C10H14	134.10955	119	2.79	4.1
13.	7-Hexadecenal	11.88	C16H30O	238.229666	57	6.29	8.3
14.	n-Nonadecanol-1	14.61	C19H40O	284.307917	71	7.4	11.5
15.	1-Decanol, 2-hexyl-	19.37	C16H34O	242.260965	57	8.08	12.2
16.	2-Hexyl-1-octanol	23.51	C14H30O	214.229666	57	5.47	7.23
17.	Carbonic acid, decyl hexadecyl ester	27.22	C27H54O3	426.407295	57	2.17	3.21

Figure 1. Antiangiogenic effects of A. maculatum extracts against EA.hy926 cells after 72 h. exposure (МТТ-assay).

Table 2. Antiproliferative effects of the tested A. maculatum extracts against EA.hy926 vascular endothelial cells (MTT-dye reduction assay).

Treatment series	GI50 (µg/mL)	TGI	LC50
A. maculatum extract (30% EtOH)	152 ± 21	338 ± 19	>1000
A. maculatum extract (70% EtOH)	277 ± 17	489 ± 23	>1000
Hydroxyarthemisinine (ART-OH)*	0.87 ± 0.1	4.06 ± 0.4	>14

*Positive control.

Figure 2. Inhibition of collagenase activity by A. maculatum extracts.

Table 3. Inhibitory activity of the tested extracts on bacterial collagenase.

Treatment series	IC50 (µg/mL)
A. maculatum extract (30% EtOH)	318.1 ± 14
A. maculatum extract (70% EtOH)	548.6 ± 27
1.10-phenanthroline*	21.17 ± 3.9

^* Positive control.

Table 4. Inhibitory effects of the tested extracts on cyclooxygenase (COX-1 and COX-2) activities.

Treatment series	IC50 (µg/mL)
	COX-1	COX-2
A. maculatum extract (30% EtOH)	589.90 ± 23.4	499.1 ± 20.5
A. maculatum extract (70% EtOH)	978.1 ± 47.9	> 1000
Nimesulide	11.36 ± 3.2	0.42 ± 0.07
Indomethacin	5.6 ± 0.9	0.74 ± 0.09

Figure 3. Effects of A. maculatum hydro-alcoholic extracts on PHA/PMA-induced secretion of IL-2 in Jurkat E.6 cells (human Т-cell model).

Note: * Indicates statistical significance at p < 0.05 vs. PHA + PMA alone; # indicates significant difference vs. the 70% EtOH extract; paired t-test.

Figure 4. Concentration dependence of the anti-radical activity of the 30% EtOH extract determined in stable free radical-based systems.

Figure 5. Typical chemiluminescent curves obtained in the chemilumiescent model systems used for evaluation of the superoxide anion and hypochlorte scavenging activity.

Note: X-axis, time of analysis [s]; Y-axis, intensity of chemiluminescence signal - relative units [RU]. Blank samples contained PBS, the tested antioxidants, and the reaction-intitiating agents; control samples contained PBS, luminol and the reaction-initiating agents but without the tested antioxidants.

Figure 6. Effect of the 30% EtOH extract on the luminol dependent CL in model systems with different ROS: OCl¯ - system of NaOCl generated hypochlorite; O₂^•¯ - system of KO₂ produced superoxide formation.