In vivo activity assessment of some Tanacetum species used as traditional wound healer along with identification of the phytochemical profile by a new validated HPLC method

Abstract

Objectives: Tanacetum species are traditionally used as insecticide, and externally wound healer as well as for anti-inflammatory and antihistaminic properties. The in vivo wound-healing and anti-inflammatory potential of four Tanacetum species, Tanacetum argenteum (Lam.) Willd. subsp. argenteum (TA), Tanacetum heterotomum (Bornm.) Grierson (TH), Tanacetum densum (Lab.) Schultz Bip. subsp. sivasicum (TD), and Tanacetum vulgare L. (TV) was investigated.

Materials and methods: The chloroform (CHCl₃) and methanol:water (80:20) extracts were prepared from the aerial parts of each plant. For assessment of the wound-healing activity, linear incision on rats and circular excision on mice wound models were used and histopathological analyses were conducted on the tissues treated with the test materials. For the evaluation of the anti-inflammatory activity, Whittle Method based on the inhibition of the acetic acid-induced increase in capillary permeability was used. In order to elucidate the phytochemical contents of the extracts, HPLC profiles of active fractions were screened and quantitative analysis was conducted within the scope of HPLC analysis.

Results: The CHCl₃ extracts of TD, TA and TV were found to have significant wound healing activity (37.1%, 30.8% and 26.1% tensile strength; 88.05%, 72.93% and 44.88% contraction values, respectively) and anti-inflammatory activities (31.5% and 26.6% inhibition values for TD and TA). Parthenolide content of the CHCl₃ extracts of TA, TH and TV were found 242.66±1.53, 190.16±5.62 and 177.51±3.73 µg/100 mg plant material, respectively.

Conclusion: According to the results, the other secondary metabolites present in the aerial parts of the Tanacetum species possibly exerted synergistic effects on the observed healing of the wounds.

Keywords: Anti-inflammatory; Asteraceae; Excision; Incision; Tanacetum; Wound-healing.

Figure 1

Histopathological view of treated tissues; Skin sections show the hematoxylin & eosin (HE) stained epidermis and dermis in A, and the dermis stained with Van Gieson (VG) in B. The original magnification was x 100 and the scale bars represent 120 µm for figures in A, and the original magnification was x 400 and the scale bars represent 40 µm for B. Data are representative of 6 animal per group. 1) Vehicle group; 2) Negative control group (untreated group); 3) T. argenteum subsp. argenteum MeOH:H₂O extract group; 4) T. argenteum subsp. argenteum CHCl₃ extract group; 5) T. heterotomum MeOH:H₂O extract group; 6) T. heterotomum CHCl₃ extract group;7) T.densum subsp. sivasicum MeOH:H₂O extract group; 8) T.densum subsp. sivasicum CHCl₃ extract group; 9) T. vulgare MeOH:H₂O extract group; 10) T. vulgare CHCl₃ extract group; 11) Reference group (wound tissue treated with Madecassol^®); Arrows pointing events during wound healing; s: scab, u: ulcus, re: re-epithelization, f: fibroblast, c: collagen, mnc: mononuclear cells, nv: neovascularization

Figure 2

HPLC chromatogram of T. argenteum subsp. Argenteum (214 nm)