Cardiovascular disease represents a leading cause of mortality and is often characterized by the emergence of endothelial dysfunction (ED), a physiologic condition that takes place in the early progress of atherosclerosis. In this study, two cytoprotective peptides derived from blue mussel chymotrypsin hydrolysates with the sequence of EPTF and FTVN were purified and identified. Molecular mechanisms underlying the cytoprotective effects against oxidative stress which lead to human umbilical vein endothelial cells (HUVEC) injury were investigated. The results showed that pretreatment of EPTF, FTVN and their combination (1:1) in 0.1 mg/mL significantly reduced HUVEC death due to H2O2 exposure. The cytoprotective mechanism of these peptides involves an improvement in the cellular antioxidant defense system, as indicated by the suppression of the intracellular ROS generation through upregulation of the cytoprotective enzyme heme oxygenase-1. In addition, H2O2 exposure triggers HUVEC damage through the apoptosis process, as evidenced by increased cytochrome C release, Bax protein expression, and the elevated amount of activated caspase-3, however in HUVEC pretreated with peptides and their combination, the presence of those apoptotic stimuli was significantly decreased. Each peptide showed similar cytoprotective effect but no synergistic effect. Taken together, these peptides may be especially important in protecting against oxidative stress-mediated ED. 相似文献
Plant extracts contain a huge variety of pharmacologically active substances. Conventionally, various chromatographic methods must be applied several times to purify functional compounds to measure their functional activity. However, conventional purification methods are time-consuming and expensive due to the laborious purification process. Recently, a high-throughput discovery method that replaces such time-consuming purification processes was introduced; this method uses 15 T ultra-high-resolution Fourier transform ion cyclotron resonance mass spectrometry (15 T FT-ICR MS) and a high-throughput screening method. This 15 T FT-ICR MS provides unparalleled resolution and sub-ppm accuracy in mass measurements, while simultaneously detecting multiple compounds without separation. The high-throughput, simultaneous multi-component discovery method known as Scaling of Correlations between Activity and Mass Profiles (SCAMP) was used to detect functional compounds in a plant extract. We validated the performance of SCAMP using 33 fractions from antioxidant-rich mulberry ethyl acetate extract and known standard antioxidants.
Results
The mulberry fruit was first separated into 33 fractions by LC and analyzed using high-resolution mass spectrometry. The antioxidative strength of the 33 fractions and standard antioxidants was measured. To validate the efficiency of this antioxidant discovery method, correlations between the antioxidation activity profile and changes in mass intensity of components within the 33 fractions were calculated to provide relative scores for the antioxidant candidate list. Enrichment curves and area under the curve (AUC) values were then calculated to compare the performance of the methods. Using this improved scoring method, five strong antioxidants, chlorogenic acid (14.2 ng), dihydoxy quercetin (46.2 ng), rutin (154.0 ng), quercetin (71.7 ng) and luteolin (3.5 ng) in 2 kg mulberry fruit, were found within the top 20 candidates.
Conclusions
We calculated AUCs in order to compare scoring methods quantitatively. Scoring systems were compared and calculated AUCs, where the AUCs for new scoring systems (0.98 and 0.99) were higher than the previously used correlation coefficient (AUC = 0.89). Using the new scoring algorithms, we successfully enriched thirteen unknown strong antioxidant candidates in addition to known antioxidants, methyl syringin and naringenin (3.5 ng) in mulberry extract. Targeted purification of these unknown candidates will significantly reduce purification time and labor.
Platycodon grandiflorum (PG), a species of herbaceous flowering perennial plant of the family Campanulaceae, has been used as a traditional oriental medicine for bronchitis, asthma, pulmonary tuberculosis, diabetes, hepatic fibrosis, bone disorders and many others similar diseases and as a food supplement. For the primary profiling of PG gas chromatography coupled with high resolution – time of flight mass spectrometry (GC/HR-TOF MS) was used as an analytical tool. A comparison of optimal extraction of metabolites was carried out with a number of solvents [hexane, methylene chloride, methanol, ethanol, methanol: ethanol (70:30, v:v)]. In extracts with methanol: ethanol (70:30 v:v) were detected higher amounts of metabolites than with other solvents. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) plots showed significant differences between the diploid and tetraploid metabolite profiles. Extracts of tetraploid showed higher amounts of amino acids, while extracts of diploid contained more organic acids and sugars.
This study was conducted to understand the effect of N supply in autumn on its uptake and distribution in tree parts and the utilization of reserve N the following year in persimmon trees (Diospyros kaki cv. Fuyu). The treated trees received 22.5-g N each for two successive years as a 3.5% urea solution from September 18 at 5-day intervals. Trees absorbed about 30% on average of the N applied in autumn. Sixty four to 83% of the N absorbed in autumn was in perennial parts, and 65–72% of that was in roots. Total N in perennial parts of the tree fertilized with N increased by over 4.28 g while leaf N changed little during senescence, indicating that the reserve N was constituted mostly by the N absorbed in autumn. Total N in the new growth was about the same as the amount of N declined in spring from perennial parts, indicating that there was little contribution by soil N to sustain new growth. Total dry weights of new growth the following year in a with-N tree were greater by over 34 g than those in a without-N tree. 相似文献
In this study, we investigated the anti-allergic effects of 3,4-dihydroxybenzaldehyde (DHB) isolated from the marine red alga, Polysiphonia morrowii, in mouse bone-marrow-derived cultured mast cells (BMCMCs) and passive cutaneous anaphylaxis (PCA) in anti-dinitrophenyl (DNP) immunoglobulin E (IgE)-sensitized mice. DHB inhibited IgE/bovine serum albumin (BSA)-induced BMCMCs degranulation by reducing the release of β-hexosaminidase without inducing cytotoxicity. Further, DHB dose-dependently decreased the IgE binding and high-affinity IgE receptor (FcεRI) expression and FcεRI-IgE binding on the surface of BMCMCs. Moreover, DHB suppressed the secretion and/or the expression of the allergic cytokines, interleukin (IL)-4, IL-5, IL-6, IL-13, and tumor necrosis factor (TNF)-α, and the chemokine, thymus activation-regulated chemokine (TARC), by regulating the phosphorylation of IκBα and the translocation of cytoplasmic NF-κB into the nucleus. Furthermore, DHB attenuated the passive cutaneous anaphylactic (PCA) reaction reducing the exuded Evans blue amount in the mouse ear stimulated by IgE/BSA. These results suggest that DHB is a potential therapeutic candidate for the prevention and treatment of type I allergic disorders. 相似文献
This study was conducted to investigate the effect of steeping conditions of waxy rice, temperatures (15, 25, and 35°C) and time periods (1, 11, and 21 days) on the expansion ratio of gangjung (a traditional Korean oil‐puffed rice snack). Physicochemical properties of waxy rice flour steeped under various conditions and expansion properties of gangjung made of the steeped waxy rice flour were investigated, and multiple regression analyses were applied between those properties to identify major physicochemical factors that optimally predict the expansion ratio of gangjung. As steeping temperature and time periods of waxy rice changed from the lowest to the highest, the expansion ratio of gangjung markedly increased (from 1,022 to 2,533%). Yet, the expansion ratio of the waxy rice sample steeped for 11–21 days at 35°C was not significantly different from the sample steeped for 21 days at 25°C, indicating that the lengthy steeping process for gangjung making can be shortened by increasing the steeping temperature. Physicochemical properties include moisture (γ = 0.85), protein (γ = –0.91), ash (γ = –0.84), potassium (γ = –0.89), magnesium (γ = –0.88), phosphorous (γ = –0.91), peak viscosity (γ = 0.77), and breakdown (γ = 0.94) of steeped waxy rice flour. These properties were highly correlated with expansion ratio (P < 0.01). Multiple regression analysis showed that the expansion ratio of gangjung was predicted successfully by the phosphorous content and breakdown value of steeped waxy rice flour. 相似文献
