Cytotoxic triterpene and steroidal glycosides from the seeds of Digitalis purpurea and the synergistic cytotoxicity of steroidal glycosides and etoposide in SBC-3 cells.

The phytochemical investigations of the seeds of Digitalis purpurea have revealed their richness in cardenolide and pregnane glycosides exhibiting potent cytotoxicity; further chemical examinations of the D. purpurea seeds have achieved the isolation of six triterpene glycosides (1-6), six spirostanol glycosides (7-12), and three furostanol glycosides (13-15), including seven previously unidentified compounds (1-3, 10-12, and 14). Here, the structures of 1-3, 10-12, and 14 were determined via extensive spectroscopic analyses, including two-dimensional (2D) NMR; hydrolysis, followed by chromatographic and spectroscopic analyses; and X-ray crystallographic analysis. The cytotoxic activities of the isolated compounds (1-15) against SBC-3 small cell lung carcinoma and TIG-3 normal human diploid fibroblast cells were evaluated. Triterpene glycoside 3 and spirostanol glycoside 9 exhibited considerable cytotoxicity with IC50 values of 1.0 and 1.7 µM, respectively; they induced apoptotic cell death, which was accompanied by the activation of caspase-3 in SBC-3 cells. Spirostanol glycoside 7 exhibited cytotoxicity toward the SBC-3 cells (IC50 1.3 µM). Additionally, 7 at 0.1 and 1.0 µM synergistically enhanced the cytotoxicity of etoposide against SBC-3 cells; compound 7 induced the release of DAMPs; the release of HMGB1, the secretion of ATP, and the exposure of CALR in the SBC-3 cells. Furthermore, the combination of 7 and etoposide resulted in increasing the extracellular release of DAMPs. These data indicated that 7, as well as its combination with etoposide, might potentially cause immunogenic cell death.

Benzofuran and coumarin derivatives from the root of Angelica dahurica and their PPAR-γ ligand-binding activity.

A phytochemical investigation of the root of Angelica dahurica led to the isolation of benzofuran and coumarin derivatives. This is the first report of the isolation and identification of three furanocoumarin sulfates from A. dahurica root. The structures of a total of twelve undescribed compounds were determined by extensive spectroscopic analysis, including 2D NMR data, hydrolysis, and solvolysis, followed by either physicochemical and spectroscopic data or X-ray crystallographic analysis. The isolated compounds were evaluated for their PPAR-γ ligand-binding activity, and six compounds showed significant PPAR-γ ligand-binding activity. In particular, the undescribed benzofuran derivative, 3-[6,7-furano-9-hydroxy-4-(2″,3″-dihydroxy-3″-methylbutyloxy)]-phenyl propionic acid, exhibited the most potent PPAR-γ ligand-binding activity and accumulated intracellular lipid in 3T3-L1 cells.

The effect of methanol extracts of tsao-ko (Amomum tsao-ko Crevost et Lemaire) on digestive enzyme and antioxidant activity in vitro, and plasma lipids and glucose and liver lipids in mice.

Our previous study showed that tsao-ko intake can lower plasma and liver triacylglycerol (TG) concentrations and has hypoglycemic and antioxidant activity in mice. This study involved separating two major fractions (A and B) from the methanol extracts (MeX) of tsao-ko using silica gel column chromatography, and then determining the effect of the fractions in vivo and in vitro to clarify the most effective components of tsao-ko. An intake of MeX and A fraction statistically significantly reduced body lipids and plasma thiobarbitutic acid reactive substances (TBARS) concentrations compared with the control and inhibited lipase and alpha-glucosidase activities. These reductions were not observed in mice fed the B fraction and these inhibitions of B fraction were mild compared with MeX and A fraction. The plasma and liver TG concentrations of each fraction group did not show significant differences compared with the control. The [M-H](+) and maximum UV absorption of the A fraction were 291 m/z and 279 nm, respectively. The peak of A fraction appeared at a similar time to the epicatechin standard in the LC/MS/MS analysis and the MS/MS spectrum of the A fraction was similar to that of the epicatechin standard. It was concluded that the most effective component of tsao-ko for body lipid reduction and hypoglycemic and antioxidant activity was contained in the polar fraction and the evidence suggested that this component could be epicatechin. However, the strongest TG lowering components of tsao-ko may be methanol insoluble.

Effect of lipid extracted from tsao-ko (Amomum tsao-ko Crevost et Lemaire) on digestive enzyme activity, antioxidant activity, plasma and liver lipids, and blood glucose levels of mice.

Lipids extracted from tsao-ko were separated into three fractions with silica gel column chromatography and fed to mice (3 mo old) for 90 d to clarify their inhibitory activity on digestive enzyme activity. The diets contained the following: control--no tsao-ko, 0.05% total lipid of tsao-ko (TL), 0.0109% chloroform fraction (CF), 0.0245% acetone fraction (AF), or 0.00365% methanol fraction (MeF). Although CF and AF slightly inhibited the activities of alpha-glucosidase, alpha-amylase, and lipase, intakes of these fractions had little influence on plasma and liver lipid concentrations when compared with the control diet. MeF did not inhibit alpha-glucosidase but had DPPH radical scavenging activity and the mice fed this fraction had the most marked reduction in plasma glucose and TBARS concentrations compared with the other diet groups. These results suggest that the fat-soluble polar components of tsao-ko contain an active component that might be associated with decreased plasma glucose and TBARS concentrations in mice.

Analysis of stress distribution in the humeroradial joint.

Stress distribution in the humeroradial joint was analyzed with pressure-sensitive film, a tactile sensor, and by the three-dimensional finite element method. Fifteen cadaveric elbows with minimal osteoarthritic changes were loaded perpendicular to the articular surface of the radial head in the full pronation, supination, and neutral positions from 0 degrees to 90 degrees. Finite element analysis of stress distribution in the joint was based on a model of the same conditions. The patterns of stress distribution were similar with all three analysis methods. High stress was concentrated laterally in supination, and medially in the neutral and pronation positions. The results of the analyses closely resembled those found in some studies of the pathophysiology of degenerative changes in the humeroradial joint.