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. 2023 Apr 9;12(8):1591.
doi: 10.3390/plants12081591.

Antitumor and Antioxidant Activities of In Vitro Cultivated and Wild-Growing Clinopodium vulgare L. Plants

Maria Petrova  1 , Lyudmila Dimitrova  1 , Margarita Dimitrova  1 , Petko Denev  2 , Desislava Teneva  2 , Ani Georgieva  3 , Polina Petkova-Kirova  4 , Maria Lazarova  4 , Krasimira Tasheva  1
Affiliations

Antitumor and Antioxidant Activities of In Vitro Cultivated and Wild-Growing Clinopodium vulgare L. Plants

Maria Petrova et al. Plants (Basel). .

Abstract

Clinopodium vulgare L. is a valuable medicinal plant used for its anti-inflammatory, antibacterial and wound-healing properties. The present study describes an efficient protocol for the micropropagation of C. vulgare and compares, for the first time, the chemical content and composition and antitumor and antioxidant activities of extracts from in vitro cultivated and wild-growing plants. The best nutrient medium was found to be Murashige and Skoog (MS) supplemented with 1 mg/L BAP and 0.1 IBA mg/L, yielding on average 6.9 shoots per nodal segment. Flower aqueous extracts from in vitro plants had higher total polyphenol content (29,927.6 ± 592.1 mg/100 g vs. 27,292.8 ± 85.3 mg/100 g) and ORAC antioxidant activity (7281.3 ± 82.9 µmol TE/g vs. 7246.3 ± 62.4 µmol TE/g) compared to the flowers of wild plants. HPLC detected qualitative and quantitative differences in phenolic constituents between the in vitro cultivated and wild-growing plants' extracts. Rosmarinic acid was the major phenolic constituent, being accumulated mainly in leaves, while neochlorogenic acid was a major compound in the flowers of cultivated plants. Catechin was found only in cultivated plants, but not in wild plants or cultivated plants' stems. Aqueous extracts of both cultivated and wild plants showed significant in vitro antitumor activity against human HeLa (cervical adenocarcinoma), HT-29 (colorectal adenocarcinoma) and MCF-7 (breast cancer) cell lines. The best cytotoxic activity against most of the cancer cell lines, combined with the least detrimental effects on a non-tumor human keratinocyte cell line (HaCaT), was shown by the leaf (250 µg/mL) and flower (500 µg/mL) extracts of cultivated plants, making cultivated plants a valuable source of bioactive compounds and a suitable candidate for anticancer therapy.

Keywords: HPLC; antitumor activity; micropropagation; polyphenols; wild basil.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Micropropagation of Clinopodium vulgare L. on MS medium with different PGRs: (a) control MS medium, (b) B1, (c) Z1, (d) K1 and (e) iP1.
Figure 2
Figure 2
In vitro cultivation of Clinopodium vulgare L.: (a) shoot multiplication on MS medium supplemented with B1I0.1, (b) in vitro rooting on half-strength MS medium and (c) plants cultivated in the experimental field.
Figure 3
Figure 3
Effects of C. vulgare extracts on the cell viability of HeLa, HT-29, MCF-7 and HaCaT cell lines as assessed by MTT test after 24 and 48 h of exposure.
Figure 4
Figure 4
Cytomorphological alterations induced by the extract of in vitro cultures of C. vulgare in HT-29 colorectal carcinoma cells. (a,c) Control untreated cells; (b,d) cells treated with C. vulgare extract. (a,b) AO/EB staining; (c,d) DAPI staining; fluorescence microscopy; objective 40×.
See this image and copyright information in PMC

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