Mitochondrial Disruption Is Involved in the Effect of Nuclear Factor of Activated T cells, Cytoplasmic 4 on Aggravating Cardiomyocyte Hypertrophy.

ABSTRACT: Nuclear factor of activated T cells, cytoplasmic 4 (NFATc4), a nuclear transcription factor, has been implicated in cardiac hypertrophy through the enhancement of hypertrophic gene expression. However, the role of NFATc4 in mitochondrial modulation is mostly unknown. The current study aimed to investigate the role of NFATc4 in regulating mitochondrial function during phenylephrine (PE)-induced cardiac hypertrophy. Our results showed that overexpression of NFATc4 aggravated the PE-induced decrease in mitochondrial genesis, membrane potential, and mitochondrial gene expression as well as impaired mitochondrial respiration. However, knockdown of NFATc4 relieved PE-induced perturbations in mitochondria and cardiomyocyte hypertrophy. Mechanistically, by activating phosphoinositide-dependent kinase 1 and promoting a combination of AKT and phosphoinositide-dependent kinase 1, phosphorylation and sequential acetylation of PGC-1α were aggravated by NFATc4 and suppressed the activity of PGC-1α. In conclusion, NFATc4-regulated factors were shown to be associated with mitochondrial function and exacerbated PE-induced mitochondrial dysfunction. These findings revealed new roles of NFATc4 in cardiac hypertrophy.

Two New Chromone Glycosides from the Roots of Saposhnikovia divaricata.

Five chromone glycosides were isolated from the water-soluble portions of 70% EtOH extract of the roots of Saposhnikovia divaricata, including two new chromone glycosides 1 and 2. The structures of the chromone glycosides were identified as (3'S)-3'-O-ß-d-apiofuranosyl-(1 → 6)-ß-d-glucopyranosylhamaudol (1), (2'S)-4'-O-ß-d-apiofuranosyl-(1 → 6)-ß-d-glucopyranosylvisamminol (2), 3'-O-glucopyranosylhamaudol (3), 4'-O-ß-d-glucopyranosylvisamminol (4), and 4'-O-ß-d-glucopyranosyl-5-O-methylvisamminol (5) on the basis of extensive spectroscopic methods, and the absolute configurations of the new compounds were elucidated by the electronic circular dichroism (ECD) calculation and acid hydrolysis. The cytotoxic activities of the glycosides 1 - 5 against three human cancer cell lines (PC-3, SK-OV-3, and H460) were evaluated. The result showed that compounds 1 - 5 had weak cytotoxic activities against the human cancer cell lines with IC50 values in the range of 48.54 ± 0.80 - 94.25 ± 1.45 µm.

Modeling and Optimum Extraction of Multiple Bioactive Exopolysaccharide from an Endophytic Fungus of Crocus sativus L.

BACKGROUND: Crocus sativus L. (saffron) is a scarce plant that has been used as food flavoring agent, coloring agent, and traditional herbal medicine. METHODS: The bioactivity of exopolysaccharide (EPS) extracted from an endophytic fungus of C. sativus was examined for the first time by antioxidative, antitumor, and antibacterial assays. The extraction conditions for EPS were optimized by combining the response surface methodology with Box-Behnken design. RESULTS: EPS exhibited excellent scavenging activities against 1,1-diphenyl-2-picrylhydrazyl, hydroxyl and superoxide anion radicals, and moderate cytotoxicities against K562, A549, HL-60, and HeLa cells. The optimum extraction conditions for EPS were as follows: precipitation time of 16 h, precipitation temperature of 3.7°C, pH 7.2, and ratio of ethanol to fermented broth of 5:1 (L/L). Under the optimized conditions, the yield of EPS reached 162 ± 6 µg/L which was close to the predicted one (165 µg/L). Moreover, high-performance liquid chromatography of monosaccharide composition showed that EPS comprised mannose, glucose, galactose xylose, and arabinose in a molar ratio of 25.6:16.5:1.0:3.8:5.4. CONCLUSION: EPS may be an eligible substitute for C. sativus and a potential bioactive source applicable to pharmaceutical and food industries. SUMMARY: Exopolysaccharide (EPS) from endophytic fungus of Crocus sativus was studied for the first timeEPS extraction was optimized by combining response surface methodology with Box-Behnken designMonosaccharide composition and EPS structure were identified by high-performance liquid chromatography and Fourier-transform infrared spectroscopy. Abbreviations used: EPS: Exopolysaccharide, RSM: Response surface methodology, BBD: Box-Behnken design, DPPH: 1,1-diphenyl-2-picrylhydrazyl, VC: Ascorbic acid, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, LB: Luria Bertani, DMSO: dimethyl sulfoxide, PMP: 1-phenyl-3-methyl-5-pyrazolone, FT-IR: Fourier transform-infrared, HPLC: High-performance liquid chromatography, 3D: Three-dimensional, 2D: Two-Dimensional.

Bioassay- and liquid chromatography/mass spectrometry-guided acetylcholinesterase inhibitors from Picriafel-terrae.

BACKGROUND: Picria fel-terrae is a traditional Chinese medicine. MATERIALS AND METHODS: A new approach to the search for acetylcholinesterase (AChE) inhibitors from Picria fel-terrae is presented. RESULTS: Bioassay- and LC-MS-guided fractionation of the ethyl acetate extract was from traditional Chinese medicine P.fel-terrae. Following primary extraction, the ethyl acetate extracts fraction of P.fel-terrae showed strong AChE inhibitory activities. So the sample was separated using highperformance liquid chromatography (HPLC). The effluent was split towards two identical 96-well fraction collectors, and the presence of the biologically interesting portion and chromatographic fractions could be readily detected by analyzing selected ion chromatograms through an electrophoresis-electrospray ionization mass spectrometry (ESIMS) system for accurate mass measurement. One 96-well plate was used for a bioassay (AChE-inhibitory assay) and detected the bioactivity and position of the relevant peak in the chromatogram. The positive well in the second 96-well plate was used for identification by LC-(+) ESIMS. CONCLUSION: As abovementioned, the AChE inhibitory constituents from P.fel-terrae by LC-bioassay-ESIMS were rapid identified. Liquid chromatography/ mass spectrometry (LC-MS) screening detected the presence of six active compounds, identified as picfeltarraenin IA (1), picfeltarraenin IB (2), picfeltarraenin IV (3), picfeltarraenin X (4), picfeltarraenin XI (5), and one unknown compound. The structures were further determined by 13C NMR. The six compounds expressed stronger AChE inhibition than the known AChE inhibitorTacrine. Above all, the value of this LC-bioassay-ESIMS methodology is highlighted by the finding and structure elucidation of the active constituents from many other structural families of natural products.