Comparative Studies on Phenolic Composition, Antioxidant, Wound Healing and Cytotoxic Activities of Selected Achillea L. Species Growing in Turkey

Abstract

Turkey is one of the most important centers of diversity for the genus Achillea L. in the world. Keeping in mind the immense medicinal importance of phenols, in this study, three species growing in Turkey, A. coarctata Poir. (AC), A. kotschyi Boiss. subsp. kotschyi (AK) and A. lycaonica Boiss. & Heldr. (AL) were evaluated for their phenolic compositions, total phenolic contents (TPC), antioxidant properties, wound healing potencies on NIH-3T3 fibroblasts and cytotoxic effects on MCF-7 human breast cancer cells. Comprehensive LC-MS/MS analysis revealed that AK was distinctively rich in chlorogenic acid, hyperoside, apigenin, hesperidin, rutin, kaempferol and luteolin (2890.6, 987.3, 797.0, 422.5, 188.1, 159.4 and 121.2 µg analyte/g extract, respectively). The findings exhibited a strong correlation between TPC and both free radical scavenging activity and total antioxidant capacity (TAC). Among studied species, the highest TPC (148.00 mg GAE/g extract) and TAC (2.080 UAE), the strongest radical scavenging (EC50 = 32.63 μg/mL), the most prominent wound healing and most abundant cytotoxic activities were observed with AK. The results suggested that AK is a valuable source of flavonoids and chlorogenic acid with important antioxidant, wound healing and cytotoxic activities. These findings warrant further studies to assess the potential of AK as a bioactive source that could be exploited in pharmaceutical, cosmetics and food industries.

Keywords: Achillea; LC-MS/MS; antioxidant; breast cancer; cytotoxic; natural product; phenolics; wound healing.

Figure 1

LC-MS/MS chromatograms of 250 ppb standard mix. Notes: Chromatographic conditions were given in Experimental section. Standard compounds: 1: quinic acid, 2: malic acid, 3: tr-aconitic acid, 4: gallic acid, 5: chlorogenic acid, 6: protocatechuic acid, 7: tannic acid, 8: tr-caffeicacid, 9: vanillin, 10: p-coumaric acid, 11: rosmarinic acid, 12: rutin, 13: hesperidin, 14: hyperoside, 15: 4-OH benzoic acid, 16: salicylic acid, 17: myricetin, 18: fisetin, 19: coumarin, 20: quercetin, 21: naringenin, 22: hesperetin, 23: luteolin, 24: kaempferol, 25: apigenin, 26: rhamnetin, 27: chrysin.

Figure 2

LC-MS/MS chromatograms of AC. Notes: Chromatographic conditions were given in Experimental Section.

Figure 3

LC-MS/MS chromatograms of AK. Notes: Chromatographic conditions were given in the Experimental Section.

Figure 4

LC-MS/MS chromatograms of AL. Notes: Chromatographic conditions were given in the Experimental Section.

Figure 5

Effects of TECA, AC, AK and AL extracts on cell proliferation. (A) Total number of NIH-3T3 fibroblasts; (B) Total number of fibroblasts proliferating by mitosis Data are mean ± SEM values (cells/mm², n = 5 in each case). * p < 0.05, ** p < 0.01, *** p < 0.001, significantly different from control values.

Figure 6

Effects of TECA, AC, AK and AL extracts on percentages of NIH-3T3 fibroblasts in fusiform (A); and polygonal (B) shapes. Data are mean ± SEM values (n = 5, in each case). * p < 0.05, ** p < 0.01, *** p < 0.001, significantly different from control values.

Figure 7

Effects of TECA, AC, AK and AL extracts on percentages of NIH-3T3 fibroblasts in round shape (A); and vacuole containing NIH-3T3 fibroblasts (B) Data are mean ± SEM values (n = 5, each case). * p < 0.05, ** p < 0.01, *** p < 0.001, significantly different from control values.

Figure 8

Effects of TECA, AC, AK and AL extracts on the number of collagen granules in NIH-3T3 fibroblasts. Data are mean ± SEM values (n = 10 in each case). *** p < 0.001, significantly different from control values.

Figure 9

Cytotoxic effects of AC, AK and AL on MCF-7 cells estimated by the MTT reduction assay. Cells were treated with nothing (control, only medium), vehicle (DMSO, 0.1% in medium), and different concentrations of the extracts for 24 h (A); and 48 h (B). Data are mean ± SEM values (n = 8).