Antioxidant Activity of Sprouts Extracts Is Correlated with Their Anti-Obesity and Anti-Inflammatory Effects in High-Fat Diet-Fed Mice.

Obesity is closely associated with oxidative stress and chronic inflammation leading to related metabolic diseases. Some natural extracts or polyphenols reportedly possess anti-obesity and anti-inflammatory effects as well as antioxidant activity. In this study, we assessed the correlations between the antioxidant, anti-obesity, and anti-inflammatory activities of plant extracts with potent antioxidant activity in diet-induced obese mice. Sprouts of Cedrela sinensis (CS) and Oenothera biennis L. (OB) were selected as the most potent antioxidant plant based on analysis of in vitro antioxidant activity of the extracts of ten different edible plants. C57BL/6 mice were fed with a high-fat diet (HFD) and orally treated with 50% ethanol extract of CS or OB at 50 or 100 mg/kg body weight 5 days a week for 14 weeks. Body weight gain, weight of adipose tissue, adipocyte size, and levels of lipid metabolism, inflammation, and oxidative stress markers were investigated. The CS or OB extract reduced body weight gain, visceral adipose tissue weight, adipocyte size, and plasma leptin levels, and expressions of adipogenic genes (PPARγ and fatty acid synthase) in the adipose tissue and liver of HFD-fed mice. Both extracts also reduced mRNA levels of pro-inflammatory cytokines (IL-6 and TNF-α) and oxidative stress-related genes (heme oxygenase- (HO-) 1 and p40phox). Body weight gain of mice was significantly correlated with visceral adipose tissue weight and adipocyte size. Body weight gain and adipocyte size were significantly correlated with plasma total cholesterol and 8-epi PGF2α levels, mRNA levels of leptin, HO-1, p40phox, and CD-11 in the adipose tissue, and mRNA levels of TNF-α in the adipose tissue and liver. These results suggest that the CS and OB extracts with potent antioxidant activity may inhibit fat deposition in adipose tissue and subsequent inflammation.

Rosa multiflora Thunb Flower Extract Attenuates Ultraviolet-Induced Photoaging in Skin Cells and Hairless Mice.

Ethanol extract (RET) of Rosa multiflora Thunb flowers and its subfractions in ethylacetate (REA) or n-butanol subfractions (RBT) were reported to have potent antioxidative and anti-inflammatory activities. In this study, we investigated if those Rosa multiflora flower (RMF) extracts prevent ultraviolet (UV)-induced biochemical damages leading to photoaging. In keratinocyte or dermal fibroblasts, RET, REA, and RBT treatments with UV irradiation significantly decreased reactive oxygen species (ROS), interleukin (IL)-6, IL-8, and matrix metalloproteinase (MMP)-1 levels through suppression of nuclear factor kappa B and mitogen-activated protein kinases. In the animal experiment, mice were orally supplemented with RET (RET group) or REA and RBT mixture (RM group) for 10 weeks, concomitantly with UV exposure. Tumor necrosis factor alpha production and MMP-13 expression were reduced in the mouse skin of RET and RM groups compared with those in the UV control (UVC) group. UV-induced IL-6 production and epidermal thickening were reduced in RM group compared with those in UVC group. Eight phenolic compounds, including quercitrin (quercetin-3-O-rhamnoside), were identified in RMF extracts. Quercitrin treatment to dermal fibroblasts significantly attenuated an increase of MMP-1 expression and a decrease of type I procollagen expression caused by UV. Collectively, RMF extracts showed protective effects from UV-induced photoaging in the skin through suppression of ROS generation, proinflammatory cytokine production, and MMP expression. Quercitrin is suggested to be one of the effective compounds.

Doenjang, A Korean Traditional Fermented Soybean Paste, Ameliorates Neuroinflammation and Neurodegeneration in Mice Fed a High-Fat Diet.

Obesity is considered a risk factor for neurodegeneration. Because fermentation of soybean increases contents of various bioactive compounds with anti-obesity and anti-diabetic activities, we investigated the protective effect of doenjang, a Korean traditional fermented soybean paste, against neuroinflammation and neurodegeneration in the cortex and hippocampus of mice fed a high-fat (HF) diet. C57BL/6J mice were fed a low-fat diet, an HF diet, an HF-containing steamed soybean diet, or an HF-containing doenjang (DJ) diet for 11 weeks. Doenjang consumption alleviated hippocampal neuronal loss, which was increased by the HF diet. Accordingly, we observed higher cell proliferation and neurotrophic factor mRNA levels in the DJ group. Contents of oxidative metabolites and mRNA levels of oxidative stress- and neuroinflammation-related genes were lower in the DJ group compared to the HF group. Dietary doenjang reduced ß-amyloid peptide (Aß) levels by regulating gene expressions involved in Aß production and degradation. Furthermore, doenjang consumption reduced tau hyperphosphorylation induced by HF feeding. Overall, doenjang was more effective than steamed soybean in suppressing neuroinflammation and neurodegeneration in mice fed an HF diet. These results suggest that bioactive compounds produced during the fermentation and aging of soybean may be involved in the enhanced neuroprotective effects of doenjang.

Topical or oral treatment of peach flower extract attenuates UV-induced epidermal thickening, matrix metalloproteinase-13 expression and pro-inflammatory cytokine production in hairless mice skin.

BACKGROUND/OBJECTIVES: Ultraviolet radiation (UV) is a major cause of skin photoaging. Previous studies reported that ethanol extract (PET) of Prunus persica (L.) Batsch flowers (PPF, peach flowers) and its subfractions, particularly the ethylacetate (PEA) and n-butanol extracts (PBT), have potent antioxidant activity and attenuate the UV-induced matrix metalloproteinase (MMP) expression in human skin cells. In this study, we investigated the protective activity of PPF extract against UV-induced photoaging in a mouse model. MATERIALS/METHODS: Hairless mice were treated with PET or a mixture of PEA and PBT either topically or orally along with UV irradiation. Histological changes and biochemical alterations of mouse skin were examined. Major phenolic compounds in PPF extract were analyzed using an ACQUITY UPLC system. RESULTS: The overall effects of topical and oral treatments with PPF extract on the UV-induced skin responses exhibited similar patterns. In both experiments, the mixture of PEA and PBT significantly inhibited the UV-induced skin and epidermal thickening, while PET inhibited only the UV-induced epidermal thickening. Treatment of PET or the mixture of PEA and PBT significantly inhibited the UV-induced MMP-13 expression, but not typeⅠ collagen expression. Topical treatment of the mixture of PEA and PBT with UV irradiation significantly elevated catalase, superoxide dismutase (SOD) and glutathione-peroxidase (GPx) activities in the skin compared to those in the UV irradiated control group, while oral treatment of the mixture of PEA and PBT or PET elevated only catalase and SOD activities, but not GPx. Thirteen phytochemical compounds including 4-O-caffeoylquinic acid, cimicifugic acid E and B, quercetin-3-O-rhamnoside and kaempferol glycoside derivatives were identified in the PPF extract. CONCLUSIONS: These results demonstrate that treatment with PET or the mixture of PEA and PBT, both topically or orally, attenuates UV-induced photoaging via the cooperative interactions of phenolic components having anti-oxidative and collagen-protective activities.

Antioxidant activity and anti-inflammatory activity of ethanol extract and fractions of Doenjang in LPS-stimulated RAW 264.7 macrophages.

BACKGROUND/OBJECTIVES: Fermentation can increase functional compounds in fermented soybean products, thereby improving antioxidant and/or anti-inflammatory activities. We investigated the changes in the contents of phenolics and isoflavones, antioxidant activity and anti-inflammatory activity of Doenjang during fermentation and aging. MATERIALS/METHODS: Doenjang was made by inoculating Aspergillus oryzae and Bacillus licheniformis in soybeans, fermenting and aging for 1, 3, 6, 8, and 12 months (D1, D3, D6, D8, and D12). Doenjang was extracted using ethanol, and sequentially fractioned by hexane, dichloromethane (DM), ethylacetate (EA), n-butanol, and water. The contents of total phenolics, flavonoids and isoflavones, 2,2-diphenyl-1 picryl hydrazyl (DPPH) radical scavenging activity, and ferric reducing antioxidant power (FRAP) were measured. Anti-inflammatory effects in terms of nitric oxide (NO), prostaglandin (PG) E2 and pro-inflammatory cytokine production and inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expressions were also measured using LPS-treated RAW 264.7 macrophages. RESULTS: Total phenolic and flavonoid contents showed a gradual increase during fermentation and 6 months of aging and were sustained thereafter. DPPH radical scavenging activity and FRAP were increased by fermentation. FRAP was further increased by aging, but DPPH radical scavenging activity was not. Total isoflavone and glycoside contents decreased during fermentation and the aging process, while aglycone content and its proportion increased up to 3 or 6 months of aging and then showed a slow decrease. DM and EA fractions of Doenjang showed much higher total phenolic and flavonoid contents, and DPPH radical scavenging activity than the others. At 100 µg/mL, DM and EA fractions of D12 showed strongly suppressed NO production to 55.6% and 52.5% of control, respectively, and PGE2 production to 25.0% and 28.3% of control with inhibition of iNOS or COX-2 protein expression in macrophages. CONCLUSIONS: Twelve month-aged Doenjang has potent antioxidant and anti-inflammatory activities with high levels of phenolics and isoflavone aglycones, and can be used as a beneficial food for human health.

Inhibitory effects of Doenjang, Korean traditional fermented soybean paste, on oxidative stress and inflammation in adipose tissue of mice fed a high-fat diet.

BACKGROUND/OBJECTIVES: Doenjang, Korean traditional fermented soybean paste has been reported to have an anti-obesity effect. Because adipose tissue is considered a major source of inflammatory signals, we investigated the protective effects of Doenjang and steamed soybean on oxidative stress and inflammation in adipose tissue of diet-induced obese mice. MATERIALS/METHODS: Male C57BL/6J mice were fed a low fat diet (LF), a high-fat diet (HF), or a high-fat containing Doenjang diet (DJ) or a high-fat containing steamed soybean diet (SS) for 11 weeks. RESULTS: Mice fed a DJ diet showed significantly lower body and adipose tissue weights than those in the HF group. Although no significant differences in adipocyte size and number were observed among the HF diet-fed groups, consumption of Doenjang alleviated the incidence of crown-like structures in adipose tissue. Consistently, we observed significantly reduced mRNA levels of oxidative stress markers (heme oxygenase-1 and p40(phox)), pro-inflammatory adipokines (tumor necrosis factor alpha and macrophage chemoattractant protein-1), macrophage markers (CD68 and CD11c), and a fibrosis marker (transforming growth factor beta 1) by Doenjang consumption. Gene expression of anti-inflammatory adipokine, adiponectin was significantly induced in the DJ group and the SS group compared to the HF group. The anti-oxidative stress and anti-inflammatory effects observed in mice fed an SS diet were not as effective as those in mice fed a DJ diet, suggesting that the bioactive compounds produced during fermentation and aging may be involved in the observed health-beneficial effects of Doenjang. CONCLUSIONS: Doenjang alleviated oxidative stress and restored the dysregulated expression of adipokine genes caused by excess adiposity. Therefore, Doenjang may ameliorate systemic inflammation and oxidative stress in obesity via inhibition of inflammatory signals of adipose tissue.

HNF4α contributes to glucose formation in aged rat hepatocytes.

Aging-dependent physiological conditions are attributed to parenchymal structural changes to cellular functions in aged organisms. Compared to the young animals, the primary hepatocytes from old rats showed a higher glucose output and a higher expression of the key gluconeogenesis-regulating enzyme, phosphoenol pyruvate carboxykinase (PEPCK). The primary hepatocytes from old rats showed a higher glucose output and a higher expression of the key gluconeogenesis-regulating enzyme, phosphoenol pyruvate carboxykinase (PEPCK), compared with those from the young animals. The in situ hybridization study showed increased PEPCK mRNA expression in the aged liver tissues. The livers from old rats showed loosened hexagonal hepatic lobular structures, increased collagen accumulation, and high expression of the hypoxia marker hypoxia-inducible factor 1α (HIF1α). Hypoxia increased the PEPCK mRNA and protein expression levels in accordance with the HIF1α expression. PEPCK promoter luciferase reporter assay showed that hypoxia increased PEPCK through transcriptional activation. Furthermore, the hepatocyte nuclear factor α (HNF4α) protein, but not the HNF4α mRNA level, increased in parallel with the PEPCK mRNA expression under hypoxic conditions. Glucose production increased under hypoxic conditions, but this increment diminished by HNF4α siRNA in young hepatocytes. Moreover, increased glucose production from old rat hepatocytes was reversed by the down-regulation of HNF4α through a specific siRNA. This study suggests that the mild hypoxic conditions in response to aging-dependent hepatic structural changes may contribute to the induction of the gluconeogenic enzyme PEPCK through HNF4α protein stabilization.

Doenjang, a fermented soybean paste, decreased visceral fat accumulation and adipocyte size in rats fed with high fat diet more effectively than nonfermented soybeans.

Soybean is known to have an anti-obesity effect. We compared the anti-obesity effect of doenjang, a fermented soybean paste, with that of nonfermented soybeans in rats. Steamed soybeans and doenjang (steamed soybeans fermented and aged for 10 months) were sampled and freeze-dried. Male Sprague-Dawley rats were fed basal (BA) (5% fat), high fat (HF) (30% fat), HF+steamed soybeans (SOY), or HF+doenjang (DJ) diet ad libitum for 8 weeks. HF significantly increased body weight gain, liver weight, hepatic triglyceride (TG) and cholesterol levels, and epididymal fat pad weight compared with BA. Compared with HF, body weight gain and hepatic TG and cholesterol levels were significantly lower in SOY and DJ groups, but they were not significantly different from each other. DJ significantly reduced visceral fat weight and epididymal adipocyte size compared with HF, whereas SOY resulted in a mild reduction without significance. This was possibly because DJ showed lowered fatty acid synthase (FAS) activity and elevated carnitine palmitoyltransferase (CPT)-1 activity in liver tissue more than SOY. SOY and DJ did not affect serum total and high-density lipoprotein-cholesterol levels compared with HF; however, DJ significantly lowered the atherogenic index and serum leptin level. In conclusion, doenjang, a fermented soybean product, was more effective than soybeans for preventing diet-induced visceral fat accumulation, possibly because of its greater effects on CPT-1 activity stimulation and FAS activity suppression. These effects may be due in part to the higher content of aglycone isoflavones in doenjang.

Dietary source of vitamin B(12) intake and vitamin B(12) status in female elderly Koreans aged 85 and older living in rural area.

Recently, we found and analyzed vitamin B(12) in some Korean traditional plant foods which had not reported, yet. This study was to investigate vitamin B(12) intake and its dietary sources and the vitamin B(12) status in the very old elderly Koreans. We measured serum vitamin B(12) level and estimated the amounts of vitamin B(12) intake from different dietary sources in female elderly Koreans aged 85 and over who had consumed a relatively low animal traditional diet for the whole life. The average age of the subjects (n = 127) was 98.0 years (85-108 years). The assessment on energy and nutrient intake involved a one-day 24-hour recall, and serum vitamin B(12) concentration was measured by radioimmunoassay. Overall diet pattern was not different between the 85-99 yr-old group and centenarians, except centenarians were taking more dairy product. The average ratio of plant food to animal food consumption was 87.5:12.5 in weight. The average vitamin B(12) intake of our subjects was 3.2 microg/day, and 52.7% of subjects consumed under estimated average requirement, 2.0 microg/day. On dietary source, 67.3% of dietary vitamin B(12) was from meat, eggs and fishes and 30.6% was from plant foods, such as soybean-fermented foods, seaweeds, and kimchi. The average serum vitamin B(12) concentration was 450.5 pg/mL, and low serum vitamin B(12) (< 200 pg/mL) was found in 9.6% of subjects. Dietary vitamin B(12) intake was significantly lower in subjects with low serum vitamin B(12) (0.79 microg/day) than those with normal serum vitamin B(12) (3.47 microg/day). There were no significant difference in vitamin B(12) intake and its dietary sources and serum vitamin B(12) level between the 85-99 yr-old group and centenarians. In conclusion, several plant-origin foods including seaweed, soybean-fermented foods, and kimchi, may contribute significantly to good vitamin B(12) status in very old elderly Koreans.

Discovery of novel sources of vitamin b(12) in traditional korean foods from nutritional surveys of centenarians.

Human longevity can be explained by a variety of factors, among them, nutritional factor would play an important role. In our study of Korean centenarians for their longevity, the apparent nutritional imbalance in the traditional semi-vegetarian diet raised a special attention, especially on vitamin B(12) status, supplied by animal foods. Interestingly, we found that the prevalence of vitamin B(12) deficient Korean centenarians was not higher compared with those from Western nations with animal-oriented traditional foods. We assumed that there might be some unveiled sources for vitamin B(12) in the Korean traditional foods. Screening of vitamin B(12) contents has revealed that some traditional soybean-fermented foods, such as Doenjang and Chunggukjang, and seaweeds contain considerable amounts of vitamin B(12). Taken together, it can be summarized that the traditional foods, especially of fermentation, might be evaluated for compensation of the nutritional imbalance in the vegetable-oriented dietary pattern by supplying vitamin B(12), resulting in maintenance of health status.

Water dropwort (Ostericum sieboldii) and Sedum (Sedum sarmentosum) delay H(2)O(2)-induced senescence in human diploid fibroblasts.

It is known that a close relationship exists among oxidative damage, senescence, and aging. Water dropwort (Ostericum sieboldii Miq. Nakai) and Sedum (Sedum sarmentosum Bunge) are popular green vegetables in Korea and are reported to have strong antioxidative activity. We investigated whether dropwort and Sedum have the potential to prevent aging using H(2)O(2)-induced prematurely senescent human diploid fibroblasts (HDFs). Dropwort and Sedum had similarly high contents of polyphenols, but dropwort had a flavonoid content about twofold higher than that of Sedum. Exposure of young HDFs to H(2)O(2) induced G2/M cell cycle arrest, positive senescence-associated (SA) beta-galactosidase (beta-gal) staining, and elevated p53, p21, and p16 protein levels. However, cotreatment with dropwort or Sedum ethanol extract significantly lowered p53, p21, and p16 levels and intracellular reactive oxygen species levels and attenuated the cell cycle arrest compared with H(2)O(2)-alone treatment. Interestingly, the increase in p16 level was prevented more quickly and clearly by dropwort treatment than Sedum treatment. The number of SA beta-gal-positive cells at 7 days after treatment was significantly reduced in dropwort-treated cells compared to H(2)O(2) alone-treated cells, whereas it was slightly reduced in Sedum-treated cells with no significance. In conclusion, dropwort showed a potential anti-senescence activity in H(2)O(2)-treated HDFs, which might be mediated by reducing p16, p21, and p53 levels and oxidative stress.

Perilla leaf, Perilla frutescens, induces apoptosis and G1 phase arrest in human leukemia HL-60 cells through the combinations of death receptor-mediated, mitochondrial, and endoplasmic reticulum stress-induced pathways.

Since it has been reported that Perilla leaves (Perilla frutescens) have antimutagenic, antioxidant, and anti-inflammatory properties, we hypothesized that Perilla leaves may have a potential anticancer activity. Therefore, we examined the possibility that cancer cell growth is reduced by treatment with a Perilla leaf ethanol extract (PLE) using human leukemia HL-60 cells and then investigated the mechanism of the growth inhibition. We found that PLE treatment suppressed cell viability in a dose-dependent manner. Flow cytometric analysis revealed that PLE treatment caused the appearance of a sub-G1 DNA peak and induced cell cycle arrest at the G1 phase. We detected DNA ladders in PLE-treated cells by agarose gel electrophoresis, and the cleavage of pro-caspase-3 and poly(ADP-ribose) polymerase with remarkable activation of caspase-8, -9, and -3. Western blot analysis revealed dose-dependent increases in Bax and cytochrome c in cytosol fractions and decreased Bid and pro-caspase-8 and -3 in PLE-treated cells. In addition, glucose-regulated protein 78, phosphorylated eukaryotic translation initiation factor 2 subunit alpha, phosphorylated c-jun N-terminal kinase, and p21 levels were increased by PLE treatment in a dose-dependent manner, whereas the p27 level was not changed. We concluded that PLE induced apoptosis through the combinations of mitochondrial, death receptor-mediated, and endoplasmic reticulum pathways and suppressed the cell proliferation via p21-mediated G1 phase arrest in HL-60 cells.

Characterization of vitamin B12 compounds from Korean purple laver (Porphyra sp.) products.

Vitamin B(12) contents of various Korean purple laver products were determined with the microbiological vitamin B(12) assay method. Although a substantial amount (133.8 microg/100 g) of vitamin B(12) was found in dried purple laver, seasoned and toasted laver products contained lesser vitamin B(12) contents (about 51.7 microg/100 g). The decreased vitamin B(12) contents in the seasoned and toasted laver products, however, were not due to loss or destruction of vitamin B(12) during the toasting process. Silica gel 60 thin layer chromatography-bioautogram analysis indicated that all Korean laver products tested contain true vitamin B(12), but not inactive corrinoid compounds. In vitro gastrointestinal digestion experiments indicated that digestion rate of vitamin B(12) from the dried Korean purple laver was estimated to be 50% under pH 2.0 conditions (as a model of normal gastric function). These results suggest that Korean purple laver products would be excellent vitamin B(12) sources for humans, especially vegetarians.

Antioxidant, antimutagenic, and antitumor effects of pine needles (Pinus densiflora).

Pine needles (Pinus densiflora Siebold et Zuccarini) have long been used as a traditional health-promoting medicinal food in Korea. To investigate their potential anticancer effects, antioxidant, antimutagenic, and antitumor activities were assessed in vitro and/or in vivo. Pine needle ethanol extract (PNE) significantly inhibited Fe(2+)-induced lipid peroxidation and scavenged 1,1-diphenyl- 2-picrylhydrazyl radical in vitro. PNE markedly inhibited mutagenicity of 2-anthramine, 2-nitrofluorene, or sodium azide in Salmonella typhimurium TA98 or TA100 in Ames tests. PNE exposure effectively inhibited the growth of cancer cells (MCF-7, SNU-638, and HL-60) compared with normal cell (HDF) in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In in vivo antitumor studies, freeze-dried pine needle powder supplemented (5%, wt/wt) diet was fed to mice inoculated with Sarcoma-180 cells or rats treated with mammary carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA, 50 mg/kg body weight). Tumorigenesis was suppressed by pine needle supplementation in the two model systems. Moreover, blood urea nitrogen and aspartate aminotransferase levels were significantly lower in pine needle-supplemented rats in the DMBA-induced mammary tumor model. These results demonstrate that pine needles exhibit strong antioxidant, antimutagenic, and antiproliferative effects on cancer cells and also antitumor effects in vivo and point to their potential usefulness in cancer prevention.