Wound Healing Properties of Jasione montana Extracts and Their Main Secondary Metabolites.

The effects of different extracts obtained from Jasione montana L. (JM1-JM6) and their main metabolites on biological processes during wound healing were evaluated. The effect on wound closure in the scratch test was established, and collagen type I synthesis and anti-inflammatory effects were assessed by flow cytometry in a human dermal fibroblast model (PCS-201-012). Additionally, the antioxidant activity (DPPH and FRAP) and degree of inhibition of elastase participating in the proliferation processes of skin fibroblasts were determined in an in vitro model. The extracts and fractions were analyzed using high-performance liquid chromatography-photodiode array detection (HPLC-PDA) to quantitatively characterize their main polyphenolic compounds. The high antioxidant activity of the JM4-JM5 fractions correlated with the content of luteolin and its derivative 7-O-glucoside. Luteolin also showed the highest anti-elastase activity with an IC50 value of 39.93 ± 1.06 µg/mL, and its substantial content in the JM4 fraction presumably determines its activity (359.03 ± 1.65 µg/mL). At lower concentrations (<50 µg/mL) of all extracts, cell proliferation and migration were significantly stimulated after 24 h of treatment. The stimulation of cell migration was comparable with that of allantoin, which was used as a positive control. However, most of the tested extracts showed limited capacity to affect collagen type I biosynthesis. Moreover, the tested samples exhibited a complex effect on cytokine secretion, and the strongest anti-inflammatory activity through the moderation of IL-1ß, IL-6 and IL-8 was observed for JM4 and luteolin. Based on the obtained results of the quantitative analysis, the anti-inflammatory activity of JM4 may be due to the high content of luteolin. In summary, extracts from J. montana, which is flavonoid-rich, promote the viability and accelerate the migration of fibroblasts as well as moderate oxidant and inflammatory processes and elastase activity. Hence, they may be potentially useful for topical therapeutic applications to stimulate the wound healing process.

Glycerol and Natural Deep Eutectic Solvents Extraction for Preparation of Luteolin-Rich Jasione montana Extracts with Cosmeceutical Activity.

Jasione montana is a plant from the family Campanulaceae rich in phenols with health-beneficial properties such as luteolin (LUT) derivatives. In this work, a glycerol-based ultrasound-assisted extraction method was developed and optimized for in total phenol (TP) and LUT content, as well as antiradical activity (RSA). The best conditions (glycerol content, temperature, plant material weight, and ultrasonication power) for the preparation of J. montana extracts richest in TP (OPT-TP), LUT (OPT-LUT), and having the best RSA (OPT-RSA) were determined. Furthermore, numerous natural deep eutectic solvents (NADES), containing proline, glycerol, betaine, urea, and glucose were prepared and used for the extraction of J. montana. Contents of TP, LUT, and RSA in the prepared extracts were established. Antioxidant and cosmeceutical activity of the prepared extracts was tested. The OPT-TP, OPT-LUT, and OPT-RSA, as well as the most efficient NADES-based extract, PG-50-TP, were excellent antioxidants and Fe2+ ion chelators. In addition, they were potent inhibitors of collagenase and hyaluronidase, as well as good significant anti-elastase and -lipoxygenase activity. The observed antioxidant- and enzyme-inhibiting activity of J. montana extracts prepared using environmentally friendly methods and non-toxic solvents makes them promising ingredients of cosmeceutical products.

In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells.

Jasione montana L. (Campanulaceae) is used in traditional Belarusian herbal medicine for sleep disorders in children, but the chemical composition and biological activity have not been investigated. In this study, the activities of J. montana extracts, their fractions and main compounds were evaluated in amelanotic melanoma C32 (CRL-1585) cells and normal fibroblasts (PCS-201-012). The extracts and fractions were analyzed using liquid chromatography-photodiode array detection-electrospray ionization-mass spectrometry (LC-PDA-ESI-MS/TOF) to characterize 25 compounds. Further, three major and known constituents, luteolin (22) and its derivatives such as 7-O-glucoside (12) and 7-O-sambubioside (9) were isolated and identified. The cytotoxic activities against fibroblasts and the amelanotic melanoma cell line were determined using the fixable viability stain (FVS) assay. The influence of diethyl ether (Et2O) fraction (JM4) and 22 on apoptosis induction was investigated using an annexin V binding assay. The obtained results showed significant cytotoxicity of JM4 and 22 with IC50 values of 119.7 ± 3.2 and 95.1 ± 7.2 µg/mL, respectively. The proapoptotic potential after 22 treatment in the C32 human amelanotic melanoma cell line was comparable to that of vinblastine sulfate (VLB), detecting 29.2 ± 3.0% apoptotic cells. Moreover, 22 displayed less necrotic potential against melanoma cells than VLB. In addition, the influences of JM4 and 22 on the dysfunction of the mitochondrial membrane potential (MMP), cell cycle and activity of caspases 3, 8, 9, and 10 were established. The effects of JM4 on MMP change (74.5 ± 3.0% of the cells showed a reduced MMP) corresponded to the results obtained from the annexin V binding assay and activation of caspase-9. JM4 and 22 displayed a significant impact on caspase-9 (40.9 ± 2.4% of the cells contained active caspase-9 after JM4 treatment and 16.6 ± 0.8% after incubation with 22) and the intrinsic (mitochondrial) apoptotic pathway. Moreover, studies have shown that JM4 and 22 affect the activation of external apoptosis pathways by inducing the caspase-8 and caspase-10 cascades. Thus, activation of caspase-3 and DNA damage via external and internal apoptotic pathways were observed after treatment with JM4 and 22. The obtained results suggest that J. montana extracts could be developed as new topical preparations with potential anticancer properties due to their promising cytotoxic and proapoptotic potential.

Recent Trends in the Application of Chromatographic Techniques in the Analysis of Luteolin and Its Derivatives.

Luteolin is a flavonoid often found in various medicinal plants that exhibits multiple biological effects such as antioxidant, anti-inflammatory and immunomodulatory activity. Commercially available medicinal plants and their preparations containing luteolin are often used in the treatment of hypertension, inflammatory diseases, and even cancer. However, to establish the quality of such preparations, appropriate analytical methods should be used. Therefore, the present paper provides the first comprehensive review of the current analytical methods that were developed and validated for the quantitative determination of luteolin and its C- and O-derivatives including orientin, isoorientin, luteolin 7-O-glucoside and others. It provides a systematic overview of chromatographic analytical techniques including thin layer chromatography (TLC), high performance thin layer chromatography (HPTLC), liquid chromatography (LC), high performance liquid chromatography (HPLC), gas chromatography (GC) and counter-current chromatography (CCC), as well as the conditions used in the determination of luteolin and its derivatives in plant material.