HPLC Fingerprint Analysis with the Antioxidant and Cytotoxic Activities of Selected Lichens Combined with the Chemometric Calculations.

The aim of this study was to evaluate the ability of multivariate techniques to predict antioxidant and cytotoxic activity of the selected lichens from the chromatographic data. A simple and reproducible HPLC-DAD technique has been used to obtain the chromatographic fingerprint profiles. Reversed phase high performance liquid chromatography (RP-HPLC) linear gradient system with methanol, water and phosphoric acid (V) (pH 2.3) as the mobile phase was used (50 min). Principal Component Analysis (PCA) has been applied to the evaluation of the phytochemical similarity between studied samples, especially between the same species collected in various places of Poland (Cetraria islandica (L.) Ach., CI, Cladina mitis Sandst., CM, Hypogymnia physodes (L.) Nyl., HP). The ability to scavenge free radicals was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) methods and the total phenolic content was determined by Folin-Ciocalteu (F-C) test. In the case of DPPH % of inhibition was higher for selected species (Pseudevernia furfuracea (L.) Zopf, H. physodes in comparison to the literature data. The FRAP test showed that the H. physodes extract had higher ability to scavenge free radical in comparison to Cladonia furcata (Huds.) Schrader and Evernia prunastri (L.) Ach., whereas P. furfuracea extract showed higher ability than C. islandica. The high content of phenolics in P. furfuracea and H. physodes confirms their high antioxidant activity. The cytotoxic activity of studied extracts was tested by cell culture method using the human HL-60 / MX2 acute CKL-22 (CRL-2257) promyelocytic leukemia tumor cell line. The lowest values of IC50 [µg∙mL-1] were obtained for: H. physodes (HP1)-99.4; C. digitate-122.6; H. physodes (HP)-136.5, C. subulata-142.6; C. mitis-180.2.

Eleutherococcus Species Cultivated in Europe: A New Source of Compounds with Antiacetylcholinesterase, Antihyaluronidase, Anti-DPPH, and Cytotoxic Activities.

Secondary metabolites of the roots of Eleutherococcus spp. cultivated in Poland, or the bioactivity, are not fully known. The 75% methanol extracts of five Eleutherococcus spp. (E. senticosus, E. divaricatus, E. sessiliflorus, E. gracilistylus, and E. henryi) were examined for the content of polyphenols and phenolic acids as well as for antiacetylcholinesterase, antihyaluronidase, anti-DPPH∗, and cytotoxic activities. The richest in polyphenols were the roots of E. henryi (10.4 mg/g DW), while in flavonoids the roots of E. divaricatus (6.5 mg/g DW). The richest in phenolic acids occurred the roots of E. henryi [protocatechuic acid (1865 µg/g DE), caffeic acid (244 µg/g DE), and p-coumaric and ferulic acids (55 µg/g DE)]. The highest inhibition of AChE was observed for E. gracilistylus and E. sessiliflorus (32%), at the concentration of 100 µg/0.19 mL of the reaction mixture, while that of Hyal for the roots of E. henryi (40.7%), at the concentration of 100 µg/0.16 mL of the reaction mixture. Among five species tested, the E. henryi extract exhibited the strongest HL-60 cell line growth's inhibition (IC50 270 µg/mL). The extracts reduced DPPH∗ in a time-dependent mode, at the concentration of 0.8 mg/mL. After 90 min from 14.7 to 26.2%, DPPH∗was reduced. A phytochemical composition and activity of the Eleutherococcus species, cultivated in Poland, are still under research; however, on the basis of the results obtained, it may be concluded that they may become a source of phytochemicals and be useful for Europe's citizens.

Effect of sodium dichloroacetate on apoptotic gene expression in human leukemia cell lines.

BACKGROUND: Sodium dichloroacetate (DCA) is an agent with anticancer properties against solid tumors. DCA also seems to have antileukemic activity. In order to affirm it we investigate the effect of DCA on cell viability and apoptotic gene expression profiles in leukemia cell lines: CEM/C1, CCRF/CEM, HL-60, HL-60/MX2. METHODS: Cell viability was assessed by trypan blue staining. The expression of 93 genes involved in the process of apoptosis was determined by real-time PCR method using Taqman Low Density Array (TLDA). RESULTS: CEM/C1, CCRF/CEM, HL-60, HL-60/MX2 cells were exposed to DCA for 24 h. The sensitivity of each cell line to DCA is different and depends on the concentration. CEM/C1 was the most sensitive with an half-maximal inhibitory concentration (IC50) value of 30 mM, while HL-60/MX2 was the most resistant with an IC50 value of 75 mM. Exposure of leukemia cells to DCA causes differences in gene expression profiles which cannot indicate that any particular pathway of apoptosis is initiated. However, the presence of 388 statistically significant correlations between expression pattern of gens was determined. CONCLUSION: We showed that DCA causes a decrease in viability of leukemia cells. The decline depends on DCA concentration. The induction of any particular apoptosis pathway is not shown in cells after DCA treatment. For that reason, studies on the molecular mechanism of cell death after exposure to DCA should be continued.