Cuban Sugar Cane Wax Acid and Policosanol Showed Similar Atheroprotective Effects with Inhibition of LDL Oxidation and Cholesteryl Ester Transfer via Enhancement of High-Density Lipoproteins Functionality.

BACKGROUND: Cuban sugarcane wax acids (SCWA) and policosanol (PCO) are mixtures of higher aliphatic acids and alcohols, respectively, purified from sugarcane wax with different chief components. Although it has been known that they have antioxidant and anti-inflammatory activities, physiological properties on molecular mechanism of SCWA have been less studied than PCO. METHODS: In this study, we compared antiatherogenic activities of SCWA and PCO via encapsulation with reconstituted high-density lipoproteins (rHDL). RESULTS: After reconstitution, SCWA-rHDL showed smaller particle size than PCO-rHDL with increase of content. PCO-rHDL or SCWA-rHDL showed distinct inhibition of glycation with similar extent in the presence of fructose. PCO-rHDL or SCWA-rHDL showed strong antioxidant activity against cupric ion-mediated oxidation of low-density lipoproteins (LDL), and inhibition of oxLDL uptake into macrophages. Although PCO-rHDL showed 1.2-fold stronger inhibition against cholesteryl ester transfer protein (CETP) activity than SCWA-rHDL, SCWA-rHDL enhanced 15% more brain cell (BV-2) growth and 23% more regeneration of tail fin in zebrafish. CONCLUSION: PCO and SCWA both enhance the beneficial functions of HDL to maximize its antioxidant, antiglycation, and antiatherosclerotic activities and the inhibition of CETP. These enhancements of HDL functionality by PCO and SCWA could exert antiaging and rejuvenation activity.

Beta-lapachone attenuates immobilization-induced skeletal muscle atrophy in mice.

Skeletal muscle atrophy reduces quality of life and increases morbidity and mortality in patients with chronic conditions. Oxidative stress is a key factor contributing to skeletal muscle atrophy by altering both protein synthesis and protein degradation pathways. Beta-lapachone (Beta-L) is known to act as a pro-oxidant in cancer cells but suppresses oxidative stress in normal cells and tissues. In the present study, we examined whether Beta-L (100 mg/kg body weight) prevents immobilization-induced skeletal muscle atrophy in male C57BL/6N mice. Skeletal muscle atrophy was induced by immobilization of left hindlimbs for two weeks, and right hindlimbs were used as controls. The muscle weights of gastrocnemius (0.132 ±â€¯0.003 g vs. 0.115 ±â€¯0.003 g in Beta-L and SLS, respectively, p < 0.01) and tibialis anterior (0.043 ±â€¯0.001 vs. 0.027 ±â€¯0.002 in Beta-L and SLS, respectively, p < 0.001) were significantly heavier in Beta-L-treated mice than that in SLS-treated mice in immobilization group, which was accompanied by improved skeletal muscle function as tested by treadmill exhaustion and grip strength test. Immobilization increased H2O2 levels, while Beta-L treatment normalized such levels (1.6 ±â€¯0.16 µM vs. 2.7 ±â€¯0.44 µM in Beta-L and vehicle, respectively, p < 0.05). Oxidative stress makers were also normalized by Beta-L treatment. Protein synthesis signaling pathways were unaltered in the case of both immobilization and Beta-L treatment. However, protein catabolic, ubiquitin-proteasomal, and autophagy-lysosomal pathways were stimulated by immobilization and were normalized by Beta-L treatment. Upregulation of transforming growth factor ß and Smad 2/3 after immobilization was significantly diminished by Beta-L treatment. These results suggest that Beta-L attenuates the loss of muscle weight and function induced by immobilization through suppression of oxidative stress.

Quercetin-Induced AMP-Activated Protein Kinase Activation Attenuates Vasoconstriction Through LKB1-AMPK Signaling Pathway.

The vascular tone plays an important role in blood pressure and flow. It is influenced by the contraction of vascular smooth muscle cells (VSMCs), which in turn is regulated by the balance between the myosin light chain kinase (MLCK) and the phosphorylated myosin light chain (p-MLC). Quercetin is a common flavonoid which is found in many fruits and red wine. Although quercetin has been widely reported to be involved in cell proliferation, migration, and apoptosis in VSMCs, it has not yet been demonstrated whether quercetin is related to vasocontraction, a function regulated by the AMP-activated protein kinase (AMPK) signaling pathway. Accordingly, the aim of this study is to investigate the molecular mechanism through which the quercetin-activated LKB1-AMPK signaling pathway regulates the contraction of VSMCs. In cultured VSMCs, quercetin activated AMPK in a dose- and time-dependent manner. Quercetin inhibited the phenylephrine (PE)-induced expression of MLCK and p-MLC through the LKB1-AMPK signaling pathway and decreased the mRNA level of MLCK. Adenovirus-AMPK DN α1 and AMPK DN α2-transduced VSMCs displayed higher p-MLC expression. Moreover, quercetin inhibited the PE-mediated contraction in rat aorta. These data suggest that the quercetin-activated LKB1-AMPK signaling pathway regulates VSMC contraction by inhibiting MLCK and p-MLC; hence, it may be a therapeutic intervention for the treatment of cardiovascular disorders such as atherosclerosis and hypertension.

High Consumption of Iron Exacerbates Hyperlipidemia, Atherosclerosis, and Female Sterility in Zebrafish via Acceleration of Glycation and Degradation of Serum Lipoproteins.

Elevated serum iron level is linked with an increased risk of diabetes and atherosclerosis. However, the pathological mechanism by which iron affects serum lipoprotein levels is unknown. To elucidate the mechanism, a high dose of ferrous ion was applied (final 60 µM, 120 µM) to human serum lipoproteins, macrophages, and human dermal fibroblast (HDF) cells. Iron-treated lipoproteins showed loss of antioxidant ability along with protein degradation and multimerization, especially co-treatment with fructose (final 10 mM). In the presence of fructose, HDF cells showed 3.5-fold more severe cellular senescence, as compared to the control, dependent on the dosage of fructose. In macrophages, phagocytosis of acetylated low-density lipoprotein (acLDL) was more accelerated by ferrous ion, occurring at a rate that was up to 1.8-fold higher, than acLDL alone. After 24 weeks supplementation with 0.05% and 0.1% ferrous ion in the diet (wt/wt), serum total cholesterol (TC) level was elevated 3.7- and 2.1-fold, respectively, under normal diet (ND). Serum triglyceride (TG) was elevated 1.4- and 1.7-fold, respectively, under ND upon 0.05% and 0.1% ferrous ion supplementation. Serum glucose level was elevated 2.4- and 1.2-fold under ND and high cholesterol diet (HCD), respectively. However, body weight was decreased by the Fe2+ consumption. Iron consumption caused severe reduction of embryo laying and reproduction ability, especially in female zebrafish via impairment of follicular development. In conclusion, ferrous ion treatment caused more pro-atherogenic, and pro-senescence processes in human macrophages and dermal cells. High consumption of iron exacerbated hyperlipidemia and hyperglycemia as well as induced fatty liver changes and sterility along with reduction of female fertility.

Inhibitory effects of (-)-loliolide on cellular senescence in human dermal fibroblasts.

Cellular senescence influences tumor suppression and progress, tissue repair and regeneration, tissue and organismal aging, and age-related diseases. Aging intervention might be an advantageous target for prevention and treatment of diverse age-related diseases. In this study, we investigated whether (-)-loliolide purified from the crude extract of Polygonum aviculare exerted inhibitory activity against cellular senescence in human dermal fibroblasts (HDFs). (-)-Loliolide diminished senescence-associated ß-galactosidase activity (SA-ß-gal), the level of p21 protein, and the level of reactive oxygen species in senescent cells induced by adriamycin treatment. (-)-Loliolide also attenuated SA-ß-gal activity in HDFs under replicative senescence. These findings imply that (-)-loliolide rescues cellular senescence in HDFs and might be useful for the development of dietary supplements or cosmetics that ameliorate tissue aging or age-associated diseases.

Quercetin-3-O-ß-D-glucuronide isolated from Polygonum aviculare inhibits cellular senescence in human primary cells.

Cellular senescence is known to contribute to tissue aging, a variety of age-related diseases, tissue regeneration, and cancer. Therefore, aging intervention might be useful for prevention of aging as well as age-related disease. In this study, we investigated compounds from Polygonum aviculare to determine if they inhibited cellular senescence in human primary cells, human dermal fibroblasts (HDFs) and human umbilical vein endothelial cells (HUVECs). Ten compounds from P. aviculare were purified and their inhibitory effects on adriamycin-induced cellular senescence were measured by observing senescence-associated ß-galactosidase (SA-ß-gal) activity and reactive oxygen species. Among them, compound 9 (quercetin-3-O-ß-D-glucuronide) showed inhibitory effects against cellular senescence in HDFs and HUVECs treated with adriamycin. Additionally, compound 9 rescued replicative senescence in HDFs and HUVECs. These data imply that compound 9 represses cellular senescence in human primary cells and might be useful for the development of dietary supplements or cosmetics that ameliorate tissue aging or aging-associated diseases.

Inhibitory effects of juglanin on cellular senescence in human dermal fibroblasts.

Cellular senescence contributes to tissue and organismal aging, tumor suppression and progress, tissue repair and regeneration, and age-related diseases. Thus, aging intervention might be a promising target for treatment and prevention of diverse age-related diseases. In the present study, we investigated whether juglanin purified from the crude extract of Polygonum aviculare exerted inhibitory activity against cellular senescence in human dermal fibroblasts (HDFs). Juglanin decreased senescence-associated ß-galactosidase activity (SA-ß-gal) and the level of reactive oxygen species in senescent cells induced by adriamycin treatment. Juglanin also repressed SA-ß-gal activity in HDFs under replicative senescence. These results suggest that juglanin represses cellular senescence in HDFs and might be useful for the development of dietary supplements or cosmetics that alleviate tissue aging or age-related diseases.

High-performance liquid chromatographic analysis for quantitation of marker compounds of Artemisia capillaris Thunb.

Two stable high-performance liquid chromatography (HPLC) methods were developed that could quantitatively analyze 10 major marker compounds of Artemisia capillaris Thunb and could also distinguish among 'Injinho' and 'Myeon-injin' and 'Haninjin'--A. capillaris collected in autumn, A. capillaris collected in spring and A. iwayomogi, which can be misused as 'Injinho' in Korean herbal drug markets. The first HPLC method was a reversed-phase chromatography using a C18 column with an isocratic solvent system of phosphoric acid (0.05%) and acetonitrile at the flow rate of 1.0 mL/min, ultraviolet (UV) detection wavelength at 254 nm and column temperature at 40°C. Calibration and quantitation were made by using acetaminophen as an internal standard (I.S-A) and chlorogenic acid (1) was determined within 20 min. The second HPLC method was a reversed-phase chromatography using a C18 column with a gradient solvent system of phosphate buffer (0.015 M, pH 6) and acetonitrile at the flow rate of 1.0 mL/min, UV detection wavelength at 254 nm and column temperature at 40°C. Calibration and quantitation were made by using ethylparaben as an internal standard (I.S-B) and 3,5-di-O-caffeoylquinic acid (2), 3,4-di-O-caffeoylquinic acid (3), 4,5-di-O-caffeoylquinic acid (4), hyperoside (5), isoquercitrin (6), isorhamnetin 3-O-robinobioside (7), isorhamnetin-3-O-galactoside (8), isorhamnetin-3-O-glucoside (9) and scoparone (10) were determined within 60 min. Pattern recognition analysis of data from the 60 samples classified them clearly into three groups. These assay methods could be applied for QA/QC of A. capillaris and Artemisia iwayomogi.

Cortical auditory evoked potential in aging: effects of stimulus intensity and noise.

OBJECTIVE: Older adults often have more difficulty understanding speech than younger adults do, particularly in the presence of noise. Such age-related speech perception difficulties may be related to declines in central auditory processing. Additionally, it has been hypothesized that impaired auditory processing might be related to neural dysynchrony. The purpose of this study was to examine the effects of stimulus intensity and noise on the N1-P2 response in younger and older normal-hearing adults. METHODS: Eight younger and 8 older normal-hearing adults participated in this study. Brief 100-ms tones (1.0 kHz, 100-60 dB SPL) in quiet and in continuous broadband noise (70 dB SPL) were used to evoke the N1-P2 responses. The N1-P2 components were analyzed as a function of stimulus intensity in both groups. RESULTS: N1 latencies to tones in quiet for older adults were delayed only at 60 dB SPL compared with those for younger adults. Additionally, N1 latencies to tones in noise were prolonged in older adults compared with those in younger adults even at 70 dB SPL (SNR = 0). No significant age effects were observed for the P2 latencies and N1-P2 amplitudes between the groups. CONCLUSION: N1 latency to tones with lower intensity and noise were delayed in older adults compared with those in younger adults. These stimulus intensity and noise issues can affect synchronized neural activity underlying the auditory processing and may provide a partial explanation for the difficulties shown by older adults in understanding speech.

Females with angina pectoris have altered lipoprotein metabolism with elevated cholesteryl ester transfer protein activity and impaired high-density lipoproteins-associated antioxidant enzymes.

In order to investigate non-invasive biomarkers for angina pectoris (AP), we analyzed the lipid and protein composition in individual lipoproteins from females with angina pectoris (n=22) and age- and gender-matched controls (n=20). In the low-density lipoprotein (LDL) fraction, the triglycerides (TG) and protein content increased in the AP group compared to the control group. The AP group had lower total cholesterol (TC) and elevated TG in the high-density lipoprotein (HDL) fraction. In the AP group, cholesteryl ester transfer protein (CETP) activity was enhanced in HDL and LDL, while lecithin:cholesterol acyltransferase (LCAT) activity in HDL3 was almost depleted. Antioxidant activity was significantly decreased in the HDL3 fraction, with a decrease in the HDL2 particle size. In the HDL3 fraction, paraoxonase and platelet activating factor-acetylhydrolase (PAF-AH) activity were much lower and the levels of CETP and apoC-III were elevated in the AP group. The LDL from the AP group was more sensitive to cupric ion-mediated oxidation with faster mobility. In conclusion, the lipoprotein fractions in the AP group had impaired antioxidant activity and increased TG and apoC-III with structural and functional changes.

Protective constituents against sepsis in mice from the root barks of Ulmus davidiana var. japonica.

In the course of isolating preventive agents against sepsis based on the in vivo assay model, eleven known compounds, (-)-catechin (1), catechin-7-O-ß-apiofuranoside (2), catechin-7-O-α-Lrhamnopyranoside (3), catechin-3-O-α-L-rhamnopyranoside (4), catechin-7-O-ß-D-glucopyranoside (5), butyl (+)-5'-methoxyisolariciresinol-9'-O-ß-D-xylopyranoside (6), lyoniside (7), nudiposide (8), α-nigerose (9), butyl α-D-fructofuranoside (10), and procyanidin B(3) (11) were isolated from the root barks of Ulmus davidiana var. japonica. Compounds 2, 6, and 8 significantly protected against sepsis in a mouse model with survival rates of mice exposed to 10 mg/kg of LPS/D-GalN ranged from 80%-100%. Among them, 8 exhibited the most potent protective effect and decreased the plasma levels of TNF-α, IL-10 and ALT activity.

Epifriedelanol from the root bark of Ulmus davidiana inhibits cellular senescence in human primary cells.

Since cellular senescence involves organismal aging as well as diverse diseases, aging intervention might contribute to inhibit the aging process as well as aging-associated diseases. We tried to search for effective compounds from the root bark of ULMUS DAVIDIANA that are able to inhibit cellular senescence in human fibroblasts (HDFs) and human umbilical vein endothelial cells (HUVECs). Twenty-two compounds from the root bark of U. DAVIDIANA were isolated and screened for their inhibitory effects on adriamycin-induced cellular senescence by measuring senescence-associated ß-galatosidase (SA- ß-gal) activity. Among twenty-two compounds isolated, epifriedelanol (3), ssioriside (15), and catechin-7-O- ß-D-glucopyranoside (22) had inhibitory effects on adriamycin-induced cellular senescence in HDFs. Friedelin (2), epifriedelanol (3), and catechin-7-O- ß-apiofuranoside (18) were active in HUVECs. In particular, epifriedelanol (3) suppressed adriamycin-induced cellular senescence as well as replicative senescence in HDFs and HUVECs. These results suggest that epifriedelanol (3) reduces cellular senescence in human primary cells and might be used to develop dietary supplements or cosmetics that modulate tissue aging or aging-associated diseases.

Inhibition of DNA topoisomerases I and II and cytotoxicity of compounds from Ulmus davidiana var. japonica.

Twenty five compounds including ten triterpenes (1-3, 5-11), six flavonoids (12-15, 24, 25), five lignans (17, 18, 21-23), two butenyl clohexnone glycosides (19-20), one fructofuranoside (16) and one fatty acid (4) were isolated from the roots of Ulmus davidiana var. japonica. The structures of those compounds were identified by comparing their physicochemical and spectral data with those of published in literatures. All the compounds were evaluated for DNA topoisomerase inhibitory activities and cytotoxicities. Among the purified compounds, 4 and 19 showed more potent inhibitory acitivities (IC(50): 39 and 19 µM, respectively) than camptothecin, as the positive control (IC(50): 46 µM) against topoisomerase I. Compounds, 4, 10, 12, 19, 24 and 25 showed strong inhibitory activities toward DNA topoisomerase II (IC(50): 0.1, 0.52, 0.47, 0.42, 0.17 µM and 17 nM, respectively), which were more potent than that of etoposide as positive control (IC(50): 20 µM). In A549 cell line, 5 and 6 showed cytotoxicities (IC(50): 4 µM and 3 µM, respectively, with IC(50) of camptothecin as positive control: 10.3 µM). In the HepG2 cell line, 3, 5 and 7 showed cytotoxicity (IC(50): 4, 3 and 4 µM, respectively, with IC(50) of camptothecin: 0.3 µM). Compounds 6, 12 and 23 showed cytotoxicities in the HT-29 cell line (IC(50): 19, 19 and 15 µM, respectively, with IC(50) of camptothecin: 2 µM).

Ethanol and water extract of purple sweet potato exhibits anti-atherosclerotic activity and inhibits protein glycation.

It is well known that extracts of purple sweet potato (PSP) have potent antioxidant activity. However, it has not been established whether extracts of PSP inhibit oxidation of low-density lipoprotein (LDL) or protein glycation. LDL oxidation and protein glycation are well-known risk factors for chronic metabolic diseases, such as atherosclerosis and diabetes mellitus. Chopped and sliced PSP and yellow sweet potato (YSP) were extracted individually at a concentration of 1 g of PSP tuber/mL using either ethanol or water for 6 hours. The PSP ethanol extract (100-fold diluted) showed stronger radical (2,2-diphenyl-1-picrylhydrazyl radical) scavenging activity than the water extract of PSP and the ethanol extract of YSP (up to a sixfold higher activity). The ethanol extract of PSP also exhibited the highest increase in ferric reducing ability among all extracts. Cupric ion-mediated LDL oxidation was strongly inhibited by the ethanol extract of PSP, with similar potency to vitamin C treatment (final concentration, 10 mM). The PSP extract strongly inhibited fructose-mediated protein glycation as determined by fluorescence spectroscopy. The PSP extract-treated apolipoprotein (apo) A-I showed a decreased multimerization pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whereas glycated apoA-I showed the strongest multimeric band. PSP extract treatment also inhibited the uptake of oxidized LDL into human macrophage cells with suppression of malondialdehyde production in the cell culture medium. In conclusion, these results suggest that the extract of PSP can be used as a putative anti-atherosclerotic and antidiabetic agent with strong antioxidant functions. This is the first report to show the biological functions of PSP extract to treat hyperlipidemic and hyperglycemic disorders.

Association of anti-obesity activity of N-acetylcysteine with metallothionein-II down-regulation.

People with upper body or visceral obesity have a much higher risk of morbidity and mortality from obesity-related metabolic disorders than those with lower body obesity. In an attempt to develop therapeutic strategies targeting visceral obesity, depot- specific differences in the expression of genes in omental and subcutaneous adipose tissues were investigated by DNA array technology, and their roles in adipocyte differentiation were further examined. We found that levels of metallothionein-II (MT-II) mRNA and protein expression were higher in omental than in subcutaneous adipose tissues. The study demonstrates that MT-II may play an important role in adipocyte differentiation of 3T3L1 preadipocytes, and that N-acetylcysteine (NAC) inhibits the adipocyte differentiation of 3T3L1 cells by repressing MT-II in a time- and dose-dependent manner. Furthermore, the intraperitoneal administration of NAC to rats and mice resulted in a reduction of body weights, and a marked reduction in visceral fat tissues. These results suggest that MT-II plays important roles in adipogenesis, and that NAC may be useful as an anti-obesity drug or supplement.

Methyl gallate and chemicals structurally related to methyl gallate protect human umbilical vein endothelial cells from oxidative stress.

Methyl gallate (meGAL) is known as one of major antioxidants. To investigate whether meGAL protects human cells from oxidative stress, meGAL extracted from Korean medicinal plant, Cercis chinensis leaves, was primarily screened using cell viability assay against oxidative stress. Human umbilical vein endothelial cells (HUVECs) were treated with three different concentrations of meGAL for indicated time. After or during meGAL treatment, H(2)O(2) was added and incubated. meGAL showed free radical scavenging effect at low concentration (0.02 mM) and cell protective effect against H2O2-mediated oxidative stress. meGAL recovered viability of HUVECs damaged by H(2)O(2)-treatment, reduced the lipid peroxidation (LPO) and decreased the internal reactive oxygen species (ROS) level elevated by H(2)O(2)-treatment. Free radical scavenging effect of meGAL was proven to be very high. Differential display reverse transcription-PCR analysis showed that meGAL upregulated the levels of regulator of chromatin condensation 1, type 1 sigma receptor and phosphate carrier protein expressions, respectively. Based on structural similarity compared with meGAL, 14 chemicals were chosen and viability assay was performed. Four chemicals, haematommic acid (56.2% enhancement of viability), gallic acid (35.0%), methylorsellinic acid (23.7%), and syringic acid (20.8%), enhanced more potent cell viability than meGAL, which showed only 18.1% enhancement of cell viability. These results suggest that meGAL and four meGAL-related chemicals protect HUVECs from oxidative stress.

Identification of amyloid beta-peptide responsive genes by cDNA microarray technology: involvement of RTP801 in amyloid beta-peptide toxicity.

Amyloid beta-peptide (Abeta), a causative molecule in the pathogenesis of Alzheimer's disease and the main component of senile plaques, is known to be neurotoxic in vitro and in vivo. The mechanisms involved in this Ab-mediated neurotoxicity are not fully understood, although there is evidence to suggest the involvement of oxidative stress, alterations in calcium homeostasis, and/or of CDK activators. Many studies have suggested that Ab may exert its toxic effect via the activation of transcription factors. Therefore, we investigated Ab- responsive genes in human neuroblastoma CHP134 cells using 3.1K human DNA microarrays. Among the several genes overexpressed or repressed by Ab, RTP801, Hi95/sestrin 2, and stanniocalcin 2 were confirmed to be Ab-mediated overexpression in the cells by semiquantitative RT-PCR. Transient expression of the sense RTP801 gene in CHP134 cells increased sensitivity to Abeta cytotoxicity and the expression of the antisense RTP801 gene protected the cells from the Abeta toxicity. These results suggest that RTP801 might play important roles in Abeta toxicity and the pathogenesis of Alzheimer's disease.