Chun Kuang Shih

Mast cell-dependent allergic responses are inhibited by ethanolic extract of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) testa.

Allergy is an immune dysfunction caused by degranulation from mast cells in the early phase and cytokine secretion in the late phase of the cell. The purpose of this study was to investigate the effects of adlay (Jobs tears, Coix lachryma-jobi L. var. ma-yuen Stapf) testa against beta-hexosaminidase release as a marker of degranulation in rat basophilic leukemia (RBL)-2H3 cells. The ethyl acetate fraction from ethanolic extracts of adlay testa (ATE-EtOAc) exhibited potent inhibitory activity that suppressed degranulation from RBL-2H3 cells stimulated by 1 microM A23187. The 20%-80% EtOAc/Hex subfractions of ATE-EtOAc significantly inhibited histamine release with a IC(50) of 75-100 microg/mL. In addition, the ATE-EtOAc subfractions suppressed interleukin (IL)-4, IL-6, and tumor necrosis factor-alpha secretion in RBL-2H3 cells, indicating that adlay testa were able to inhibit cytokine secretion. In order to explore the inhibitory mechanism of adlay testa in mast cell degranulation, we examined the activation of intracellular signaling molecules. Adlay testa inhibited the phosphorylation ERK expression. Furthermore, the two major active compounds, 4-hydroxyacetophenone and p-coumaric acid, were isolated from the ATE-EtOAc subfractions. These results suggest that ATE had an inhibitory effect on allergic response via the ERK signaling transduction in RBL-2H3 cells.

β-Carotene and canthaxanthin alter the pro-oxidation and antioxidation balance in rats fed a high-cholesterol and high-fat diet

This study investigated the effects of beta-carotene and canthaxanthin on lipid peroxidation and antioxidative enzyme activities in rats fed a high-cholesterol, high-fat diet. Wistar rats were divided into six groups. Negative control group (group NC) received a high-fat (150 g/kg) diet; cholesterol control group (group CC) received a high-cholesterol (10 g/kg), high-fat diet. The other four groups were fed a high-cholesterol, high-fat diet supplemented with crystal beta-carotene (group BC), beta-carotene beadlet (group BB), canthaxanthin beadlet (group CX) or alpha-tocopherol (group AT). Blood and livers were collected for analysis after 6 weeks of feeding. Group BB had significantly lower hepatic thiobarbituric acid reactive substance (TBARS) and conjugated diene concentrations, whereas group CX had a significantly lower plasma TBARS concentration than did group CC. In erythrocytes, glutathione peroxidase activities were significantly greater in groups BC, BB and CX than in group CC. Moreover, compared with group CC, catalase activities were significantly greater in groups BB and CX, and superoxide dismutase (SOD) activity was significantly greater in group BB. In livers, SOD activities were significantly greater in groups BC, BB and CX, and glutathione reductase activities were significantly greater in groups BB and CX than in group CC. Compared with group CC, hepatic retinol and alpha-tocopherol concentrations were significantly greater in groups BC, BB and CX, whereas plasma and hepatic cholesterol concentrations were significantly lower in group BC. These findings suggest that beta-carotene and canthaxanthin altered the pro-oxidation and antioxidation balance and suppressed cholesterol-induced oxidative stress via modulation of antioxidant system and cholesterol metabolism.

Ethyl acetate fraction of adlay bran ethanolic extract inhibits oncogene expression and suppresses DMH-induced preneoplastic lesions of the colon in F344 rats through an anti-inflammatory pathway

Adlay ( Coix lachryma-jobi L. var. ma-yuen Stapf) is a grass crop and was reported to possess anti-inflammatory activity and an antiproliferative effect in cancer cell lines. The purpose of this study was to evaluate the effects of the ethyl acetate fraction of an adlay bran ethanolic extract (ABE-Ea) on colon carcinogenesis in an animal model and investigate its mechanism. Male F344 rats received 1,2-dimethylhydrazine (DMH) and consumed different doses of ABE-Ea. The medium-dose group (17.28 mg of ABE-Ea/day) exhibited the best suppressive effect on colon carcinogenesis and prevented preneoplastic mucin-depleted foci (MDF) formation. Moreover, RAS and Ets2 oncogenes were significantly down-regulated in this group compared to the negative control group, whereas Wee1, a gene involved in the cell cycle, was up-regulated. Cyclooxygenase-2 (COX-2) protein expression was significantly suppressed in all colons receiving the ABE-Ea, indicating that ABE-Ea delayed carcinogenesis by suppressing chronic inflammation. ABE-Ea included considerable a proportion of phenolic compounds, and ferulic acid was the major phenolic acid (5206 microg/g ABE-Ea) on the basis of HPLC analysis. Results from this study suggest that ABE-Ea suppressed DMH-indued preneoplastic lesions of the colon in F344 rats and that ferulic acid may be one of the active compounds.

Effects of adlay bran and its ethanolic extract and residue on preneoplastic lesions of the colon in rats

BACKGROUND Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) is a cereal crop used in traditional Chinese medicine and as a nutritious food. Epidemiologists have suspected that the low cancer rates in southeastern China might be related to adlay. Previous studies have shown that adlay has anti-tumour and anti-inflammatory activity. This study investigated the effect of adlay bran and its fractions on chemically induced colon carcinogenesis in rats. RESULTS Adlay bran and its ethanolic extract and residue significantly reduced the number of preneoplastic aberrant crypt foci (ACF) and modified their mucin composition. The inhibitory effect of adlay bran ethanolic extract on ACF showed a dose dependence. Adlay bran and its ethanolic extract suppressed small ACF (one, two or three crypts) and ACF in the distal colon, while the residue suppressed large ACF (four or more crypts). CONCLUSION These findings suggest the possibility that adlay bran and its ethanolic extract and residue inhibit colonic preneoplastic lesions in an early stage. Adlay and its fractions may have the potential to be developed as chemopreventive cereal products.

Application of the solvent extraction technique to investigation of the anti-inflammatory activity of adlay bran.

The current study utilised a bioassay-directed chemical analysis scheme to screen the anti-inflammatory activity of fractions and compounds from adlay bran (AB). Liquid-liquid extraction couple with liquid chromatography-mass spectrometry (LC-MS) was applied to the isolation, analysis and identification of active components in AB samples. Ethanol extracts of AB (ABE) and ethyl acetate extracts AB (ABEa) were obtained and further partitioned with different solvents. The results showed that among all 16 kinds of fractions from ABE and ABEa, ABEa-Ea-B (80% Ea/n-hexane sub-fraction from ABE-Ea) had the most potent inhibitory effects on NO production, iNOS and COX-2 expressions, and proinflammatory IL-6 and TNF-α secretion in lipopolysaccharide-activated RAW264.7 cells system. Mechanistic data from luciferase reporter-gene assay revealed that the anti-inflammatory action of ABEa-Ea-B may be associated with inhibition of NF-kB transcriptional activity. Notably, tangeretin, nobiletin, and p-hydroxybenzoic acid were found to be the main active compounds for the anti-inflammatory properties in ABEa-Ea-B.

Constituents in purple sweet potato leaves inhibit in vitro angiogenesis with opposite effects ex vivo

OBJECTIVE The aim of this study was to investigate the in vitro effect of polyphenols in purple sweet potato leaves (PSPLs) on angiogenesis in human umbilical vascular endothelial cells (HUVECs). The ex vivo effect was test in human serum collected from the subjects who consumed 200 g of PSPL in a low polyphenol diet versus a low polyphenol diet. METHODS Methanolic extract from PSPLs and human sera from subjects were treated with HUVECs and the effects of cell proliferation, migration, tube formation, and matrix metalloproteinase activity were investigated. RESULTS The PSPL polyphenols at 0.2 to 0.6 mM gallic acid equivalents inhibited proliferation, migration, and tube formation of vascular endothelial growth factor-treated HUVECs. Further, the activity of secreted matrix metalloproteinase-2 was decreased by at least 13.8%. However, 5% PSPL serum increased migration and tube formation of HUVECs by 110% and 56.9%, respectively, compared with serum from subjects on the low polyphenol diet. Further, the activity of matrix metalloproteinase-9 was increased by 128% in the PSPL serum. CONCLUSION These results suggest that PSPL polyphenols inhibited in vitro angiogenesis, but PSPL constituents might shift serum biochemistries to be more proangiogenic.

Riboflavin protects mice against liposaccharide-induced shock through expression of heat shock protein 25.

Riboflavin (vitamin B2) is a water-soluble vitamin essential for normal cellular functions, growth and development. This study aimed to investigate the effects of vitamin B2 on the survival rate, and expressions of tissue heat shock protein 25 (HSP25) and heat shock factor 1 (HSF1) in mice undergoing lipopolysaccharide (LPS) induced shock. Mice were assigned to four groups, saline vehicle, LPS, LPS plus low dose of vitamin B2 (LB2) and LPS plus high dose of vitamin B2 (HB2). Vitamin B2 (1 and 10mg/kg BW) was administered intraperitoneally at 2 and 0 h before the i.p. administration of LPS. At the end of the experiment, the survival rate monitored was 10, 20, 60, and 100% for LPS, LB2, HB2, and saline mice, respectively. HSP25 expressions in the heart and lung were significantly enhanced in a time-dependent manner in the HB2 mice as compared to the saline mice (p < 0.05), but not altered in the LB2 mice. In the HB2 mice, plasma riboflavin concentrations reached 300 nM at 6h post LPS and returned to the 0 h level at 72 h. The results showed that high dose of riboflavin could decrease LPS-induced mortality through an increased expression of HSP25.

Screening of Ethylnitrosourea Mice With Fatty Acid Oxidation Disorders by a Candidate Gene Approach After Proteome Analysis

Background/Purpose Ethylnitrosourea (ENU) is an alkylating agent and primarily induces point mutations such as AT to TA transversions and AT to GC transitions. Due to its high mutagenicity, ENU mouse mutagenesis enables the generation and identification of mouse mutants with aberrance in various phenotypes and to identify novel genes relevant for the expression of the phenotype. The purpose of this study was to investigate the candidate genes involved in fatty acid oxidation disorders by the proteomic approach. Methods We screened ENU mice from 39 families from previously published data and identified two mutant mice that had a striking elevation in blood C4-OH short chain fatty acids compared with ENU controls. Total mitochondrial proteins were extracted from the gastrocnemius for two-dimensional electrophoresis, and two downregulated proteins, adenylate kinase isoenzyme 1 (AK1) and adenosine-5′-triphosphate (ATP) synthase D chain (ATP5H), were identified in the mutant mice through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Results After genomic polymerase chain reaction and direct sequencing of Ak1 and Atp5h , no variation was found in both gene sequence analyses. Conclusion Proteomic profiling can be a useful approach for detecting dynamic protein expression in ENU-induced mice. It is important to further clarify mechanisms of the mutant C4-OH disorder responsible for this expression.