Preclinical study of the medicinal plants for the treatment of malignant melanoma.

Melanoma is the most aggressive type of skin cancer and originates from pigment-containing cells called melanocytes. The incidence of melanoma has been increasing worldwide. In the current study, the cytotoxic and photo-cytotoxic activities of different medicinal plants from Lamiaceae (Salvia cedronella, Salvia chionantha, and Salvia adenophylla), Asteraceae (Klasea kurdica, Klasea bornmuelleri, and Achillea millefolium), Apiaceae (Cuminum cyminum, and Anethum graveolens), and Polygonaceae (Rumex crispus) families were studied against HT 144 (Human malignant melanoma) cancer cell lines. The activities were performed by employing the MTT assay. Moreover, the apoptotic effects of the plant extracts were investigated by flow cytometry with annexin V/PI dual staining technique. The production of intracellular ROS by DCFH-DA technique and the effects of TNF-α secretion on apoptosis were also investigated. All plant extracts exhibited cytotoxic, and photo-cytotoxic effects against HT 144 cancer cells. Salvia species and Klasea species induced apoptosis via intracellular ROS generation secreted by TNF-α. On the other hand, A. millefolium, C. cyminum, A. graveolens, and R. crispus extracts induced apoptosis due to the intracellular generation of ROS, but, via the different pathway. In conclusion, this study indicates that the tested medicinal plant extracts have the potential in the treatment of melanoma.

The anticancer activity of visnagin, isolated from Ammi visnaga L., against the human malignant melanoma cell lines, HT 144.

Melanoma is a cancer of melanocyte cells and has the highest global incidence. There is a need to develop new drugs for the treatment of this deadly cancer, which is resistant to currently used treatment modalities. We investigated the anticancer activity of visnagin, a natural furanochromone derivative, isolated from Ammi visnaga L., against malignant melanoma (HT 144) cell lines. The singlet oxygen production capacity of visnagin was determined by the RNO bleaching method while cytotoxic activity by the MTT assay. Further, HT 144 cells treated with visnagin were also exposed to visible light (λ ≥ 400 nm) for 25 min to examine the illumination cytotoxic activity. The apoptosis was measured by flow cytometry with annexin V/PI dual staining technique. The effect of TNF-α secretion on apoptosis was also investigated. In standard MTT assay, visnagin (100 µg/mL) exhibited 80.93% inhibitory activity against HT 144 cancer cell lines, while in illuminated MTT assay at same concentration it showed lesser inhibitory activity (63.19%). Visnagin was induced apoptosis due to the intracellular generation of reactive oxygen species (ROS) and showed an apoptotic effect against HT 144 cell lines by 25.88%. However, it has no effect on TNF-α secretion. Our study indicates that visnagin can inhibit the proliferation of malignant melanoma, apparently by inducing the intracellular oxidative stress.

Effect of Sideritis leptoclada against HT-144 human malignant melanoma.

Sideritis leptoclada O. Schwarz et P.H. Davis extracts were evaluated for its singlet oxygen production capacity using spectrophotometric method. The extracts producing singlet oxygen were then evaluated for cytotoxicity against malignant melanoma cancer (HT-144) and fibroblast (3T3) cell lines using the 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. The photocytotoxicity against the HT-144 human melanoma cell line in the presence of illumination (∼≥400 nm) was also evaluated. In the standard MTT assay, the ethanol extract of S. leptoclada (100 µg/ml) showed 83.49±3.33% inhibition of HT-144 cancer cells, whereas in the illuminated MTT assay, it showed 77.46±1.97% inhibition of HT-144 cancer cells. The effects of ethanol extract on reactive oxygen species production, apoptosis, and tumor necrosis factor-α secretion were also evaluated on HT-144 cell lines. The extract triggered an increase in intracellular reactive oxygen species production and tumor necrosis factor-α secretion compared with the respective controls. Thus, the ethanol extract may cause apoptosis. The LC-MS/MS analyses of S. leptoclada ethanolic extract showed that it has quinic acid (137213±11.25 µg/g extract), malic acid (1468±0.16 µg/g extract), chlorogenic acid (881.7±0.06 µg/g extract), and apigetrin (223.2±0.13 µg/g extract) as major constituents. The ethanolic extract of S. leptoclada should be further investigated as a potential treatment for malignant melanoma cancer.

The sensitive capillary electrophoretic-LIF method for simultaneous determination of curcuminoids in turmeric by enhancing fluorescence intensities of molecules upon inclusion into (2-hydroxypropyl)-ß-cyclodextrin.

Curcuminoids have received great attention in the past decades due to their health benefit properties. The aim of this study is to develop a very simple, rapid, and sensitive capillary zone electrophoresis technique coupled with a laser induced fluorescence detector (LIF) for the simultaneous determination of three major curcuminoids of turmeric, namely, curcumin, demethoxy curcumin (DMC), and bisdemethoxy curcumin (BDMC). Background electrolyte was selected as borate at pH 9.6 and (2-hydroxypropyl)-ß-cyclodextrin (2-HP-ß-CD) was added to prevent rapid alkali degradation of curcuminoids in buffer and to increase fluorescence intensities of molecules. With the addition of 2-HP-ß-CD to the separation electrolyte, the fluorescence signal intensities of curcuminoids were enhanced considerably by 30, 40, and 54 fold for curcumin, DMC, and BDMC, respectively. The three curcuminoids of turmeric were fully separated and quantified in less than 4.5 min. The repeatability of the peak areas of curcuminoids for intra-day and inter-day experiments was in the satisfactory range of 2.26 and 2.55%, respectively. The LOD and LOQ values for the developed method were equal to or less than 0.081 and 0.270 µg/mL, respectively, for all curcuminoids. The developed method was successfully applied to find curcuminoids amount in turmeric samples and herbal supplements.

Environmental friendly alkaline sulfite anthra quinone-methonal (ASAM) pulping with Rumex crispus plant extract of woody materials.

ASAM with Rumex crispus extract organosolv pulping was developed by using 1,5-dihydroxy-3-methoxy-7-methyl-anthraquinone from Rumex crispus root, instead of anthraquinone. ASAM was also produced as a control pulping. Both pulps were made by handsheets from fast growing P. deltoides clone (Samsun p. clone), Robinia pseudoacacia L. and Pinus pinaster grown in Turkey for wood fibrous raw materials. The mechanical consisting tensile, bursting and tear values and optical values of ASAM handsheets yellowness, brightness and whiteness were compared to ASAM with Rumex crispus L. extracted. It is concluded that ASAM with Rumex crispus extract pulping suits well in the manufacturing of special papers.

Analysis of anthraquinones in Rumex crispus by micellar electrokinetic chromatography.

A micellar electrochromatographic method was performed for the analysis of the pharmaceutically important anthraquinones from the root of Rumex crispus. The separation of 1,5-dihydroxy-3-methylanthraquinone (1); 1,3,5-trihydroxy-6-hydroxymethylanthraquinone (2); 1,5-dihydroxy-3-methoxy-7-methylanthraquinone (3) was achieved in 6min using a running buffer containing 10mmol/l sodium borate, 50mmol/l sodium dodecylsulfate, and 25% acetonitrile at pH 10.6. The method is simple, rapid, and reproducible.

Phytochemical studies on Ruta chalepensis (Lam.) Lamarck.

Ruta chalepensis is a rich source of important secondary metabolites such as furanocoumarins and alkaloids. Besides, it is a medicinal plant and still used in traditional medicine. For that reason, its chemical composition, medicinal properties, and uses were reviewed in this article.

The chemical investigations on the ripe fruits of Ammi visnaga (Lam.) Lamarck growing in Turkey.

Two furanochromones and one furanochromone glycoside were isolated from the fruits of Ammi visnaga (L.) Lam. They were identified as khellin, visnagin, and khellol glycoside by interpretation of spectral analyses. Quantitative determination of furanochromones in A. visnaga (L.) ripe fruits from Hatay region (Turkey) was carried out by ultraviolet spectrophotometry and gas chromatography. In addition, photochemical properties of furanochromones and chemical composition of essential oil were determined.

1,5-dihydroxyanthraquinones and an anthrone from roots of Rumex crispus.

From the roots of Rumex crispus, two known anthraquinones and a new one together with a new anthrone were isolated and the structures of compounds 1-4 were elucidated by spectroscopic means. The singlet oxygen generation capacity was tested with 1,3-diphenylisobenzofuran (DPBF) for compounds 1-4.