Korean Scutellaria baicalensis Georgi flavonoid extract induces mitochondrially mediated apoptosis in human gastric cancer AGS cells.

Korean Scutellaria baicalensis Georgi has been widely used in Korean folk medicines for its range of medicinal benefits, including its anticancer effect. The aim of the present study was to investigate the underlying molecular mechanism of action of a flavonoid extract from Korean Scutellaria baicalensis Georgi (FSB) on AGS human gastric cancer cells (gastric adenocarcinoma) in which FSB exhibits an anticancer effect. Treatment of AGS cells with FSB significantly inhibited cell viability in a concentration-dependent manner. Furthermore, FSB significantly increased the proportion of cells in sub-G1 phase, and Annexin V and Hoechst 33258 fluorescent staining confirmed the apoptotic cell death. Furthermore, western blotting results identified that treatment of AGS cells with FSB significantly downregulated the expression of caspase family members, namely procaspases 3 and 9, and poly(ADP-ribose) polymerase (PARP), and subsequently upregulated cleaved caspase 3 and cleaved PARP. It was observed that FSB treatment significantly decreased the mitochondrial membrane potential of AGS cells. In addition, the ratio of the mitochondrion-associated proteins B cell lymphoma 2-associated X protein and B cell lymphoma extra large was upregulated. The results of the present study provide novel insight into the underlying molecular mechanism of the anticancer effects of FSB on AGS human gastric cancer cells and indicate that FSB may be an alternative chemotherapeutic agent for the treatment of gastric cancer.

Inhibitory effects on N-nitrosodimethylamine formation by decrease of salted-fermented fish products and increase of condiments in kimchi.

Ethanol extracts from developed kimchi condiments (KME, KMEE) and mixtures of sub-ingredients (ME, MEE) showed high nitrite scavenging activity. ME was able to scavenge 89% of total nitrite at 50 mg/mL ME and pH 1.2. The nitrite scavenging abilities of KME and KMEE were significantly higher than in ethanol extract from the control condiment. The inhibitory effects on N-nitrosodimethylamine (NDMA) formation by decrease of salted-fermented fish products (Jeot-gal) and increase of condiments in the composition of kimchi were investigated. The modified kimchi (KM) was prepared with new condiments, which included new sub-ingredients and reduced Jeot-gal. The NDMA and its precursor levels were significantly decreased in KM compared with those in the control kimchi (KC). The KM also obtained higher sensory scores than KC. Therefore, the increase of sub-ingredients and reduction of Jeot-gal in kimchi would be recommended for production of reduced-NDMA kimchi while maintaining or even enhancing flavor profiles.

Evaluation of phenotypic, functional and molecular characteristics of porcine mesenchymal stromal/stem cells depending on donor age, gender and tissue source.

The biological properties of mesenchymal stem cells (MSCs) are influenced by donor age, gender and/or tissue sources. The present study investigated the cellular and molecular properties of porcine mesenchymal stromal/stem cells (MSCs) isolated from different tissues (adipose & dermal skin) and sex at different ages (1 week & 8 months after birth) with similar genetic and environmental backgrounds. MSCs were analyzed for alkaline phosphatase (AP) activity, CD90 and Oct3/4 expression, in vitro differentiation ability, senescence-associated ß-galactosidase (SA-ß-Gal) activity, telomeric properties, cell cycle status and expression of senescence (IL6, c-myc, TGFß, p53 and p21)- and apoptosis (Bak and Bcl2)-related proteins. An age-dependent decline in AP activity and adipogenesis was observed in all MSCs, except for male A-MSCs. CD90 expression did not change, but SA-ß-Gal activity increased with advancement in age, except in A-MSCs. Telomeric properties were similar in all MSCs, whereas expression levels of Oct3/4 protein declined with the advancement in age. p21 expression was increased with increase in donor age. Male derived cells have shown higher IL6 expression. The expression of p53 was slightly lower in MSCs of dermal tissue than in adipose tissue. Bak was expressed in all MSCs regardless of age, but up regulation of Bcl2 was observed in DS-MSCs derived at 1 week after birth. In conclusion, adipose tissue-derived MSCs from young female individuals were found to be more resistant to senescence under in vitro culture conditions.

Aged red garlic extract suppresses nitric oxide production in lipopolysaccharide-treated RAW 264.7 macrophages through inhibition of NF-κB.

Lipopolysaccharides (LPS) activate nuclear factor kappa B (NF-κB), a transcription factor that is involved in inflammatory response. The pathways that activate NF-κB can be modulated by phytochemicals derived from garlic. We recently demonstrated that aged red garlic extract (ARGE), a new formulation of garlic, decreases nitric oxide (NO) generation by upregulating of heme oxygenase-1 (HO-1) in RAW 264.7 cells activated by LPS. However, the effects of ARGE on LPS-induced NF-κB activation are unknown. This study was performed to evaluate whether ARGE regulates LPS-induced NO production by modulation of NF-κB activation in macrophages. The inhibition of NF-κB by Bay 11-7085, an inhibitor of NF-κB, decreased LPS-induced NO production. ARGE treatment markedly reduced LPS-induced NO production and NF-κB nuclear translocation. ARGE downregulated expression of inducible nitric oxide synthase (iNOS) and upregulated expression of HO-1, a cytoprotective and anti-inflammatory protein. However, Bay 11-7085 only reduced iNOS expression. The NO production and iNOS expressions upregulated by suppression of HO-1 were suppressed by treatment with ARGE and Bay 11-7085. These results show that ARGE reduces LPS-induced NO production in macrophages through inhibition of NF-κB nuclear translocation and HO-1 activation. Compared to Bay 11-7085, ARGE may enhance anti-inflammatory effects by controlling other anti-inflammatory signals as well as regulation of NF-κB.

Aged red garlic extract reduces cigarette smoke extract-induced cell death in human bronchial smooth muscle cells by increasing intracellular glutathione levels.

Increasing antioxidant capacity has been proposed as a promising strategy to prevent cigarette smoke-induced lung diseases. This study tested whether garlic extracts prevented cigarette smoke extract (CSE)-induced cell death in human bronchial smooth muscle cells (HBSMCs). Garlic extracts were prepared from fresh raw garlic (FRG), aged black garlic (ABG) and aged red garlic (ARG). Treatment of HBSMCs with 10% CSE induced cell death accompanied by activation of caspase. Of the garlic extracts, treatment with ARG extract reduced CSE-induced cell death. The combination of ARG extract with CSE attenuated the CSE-induced reduction in glutathione (GSH) content, generation of reactive oxygen species (ROS) and induction of heme oxygenase-1 expression compared with CSE treatment without ARG extract. Furthermore, the combination of L-BSO, a GSH synthesis inhibitor, with ARG and CSE extracts failed to increase the intracellular GSH content and cell viability. Taken together, these results demonstrate that ARG extract reduces CSE-induced cell death by increasing GSH content and reducing ROS generation in HBSMCs.

Hypolipidemic and hypoglycemic effects of Orostachys japonicus A. Berger extracts in streptozotocin-induced diabetic rats.

The hypolipidemic and hypoglycemic effects of two dietary dosages (0.1% and 0.5%) of water and 80% ethanol extracts from hot-air dried Orostachys japonicus A. Berger were evaluated in the serum and organ tissues of streptozotocin-induced diabetic rats. The STZ-induced diabetic groups supplemented with the O. japonicus extracts showed significantly higher body weight compared to a diabetic control group at the end of experiment. The extracts exhibited substantial hypoglycemic effects by significant reductions of fasting blood glucose levels at all time points tested compared to the initial stage before treatment of the extracts. Declines of serum and hepatic triglyceride levels were greater than declines of total cholesterol in the groups treated with the 0.5% O. japonicus extract (DBW2 and DBE2) when compared to the DBC group. Hepatic glycogen content was higher in the groups treated with O. japonicus extract, while lipid peroxide content was decreased in these treated groups compared to the DBC group. Hepatic antioxidant activity was significantly increased in the groups supplemented with the O. japonicus ethanol extract. The hypolipidemic and hypoglycemic effects of the O. japonicus ethanol extract were significantly greater than the effects of the water extract. Based on this study, it seems that O. japonicus ethanol extract, due to its higher phenolic and flavonoid components than the water extract, may control blood glucose and alleviate hyperlipidemia in diabetes.

Antioxidative and hypocholesterolemic activities of water-soluble puerarin glycosides in HepG2 cells and in C57 BL/6J mice.

Puerarin is an isoflavone derived from Kudzu roots and has antioxidant and hypocholesterolemic effects; however, its insolubility often limits its biological availability in vivo. Using a novel transglycosylation process, the solubility of puerarin glycosides was increased >100-fold, but it was not known whether these modified puerarin glycosides maintained biological activities. We found that water-soluble puerarin glycosides fully maintained antioxidant activities compared with puerarin assessed by radical scavenging activity, reducing power assay, superoxide dismutase activity, and non-site-specific hydroxyl radical scavenging activity. Both puerarin and its glycosides also significantly reduced low-density lipoprotein (LDL) oxidation. Mice fed with puerarin glycosides (0.1% w/w) showed significantly reduced plasma total cholesterol levels, thus, we further investigated their hypocholesterolemic mechanisms by assessing several key gene expressions both in vitro and in vivo. Puerarin and its glycosides induced multiple changes in hepatic cholesterol metabolism. The LDL receptor promoter activity was increased dose-dependently in puerarin glycosides-treated HepG2 cells. Accordingly, the expression of LDL receptor mRNA and protein were also significantly increased in HepG2 cells and mouse livers. The transcription and translation of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase were down-regulated both in vitro and in vivo. The cholesterol 7alpha-hydroxylase (CYP7A1) mRNA levels were not affected in vitro but significantly up-regulated in the mouse livers. Collectively, our results show that puerarin and its glycosides are biologically fully active isoflavone and have antioxidant and hypocholesterolemic effects in HepG2 cells and in C57BL/6J mice. In the livers, hypocholesterolemic effects of puerarin glycoside may be achieved by multiple mechanisms including increasing LDL uptake, reducing cholesterol biosynthesis, and possibly enhancing cholesterol degradation.

Free radical scavenging activity of a novel antioxidative peptide isolated from in vitro gastrointestinal digests of Mytilus coruscus.

A low-molecular-weight peptide with potent antioxidative activity was obtained from Mytilus coruscus muscle protein using an in vitro gastrointestinal digestion system. The potent antioxidant peptide, which was identified as Leu-Val-Gly-Asp-Glu-Gln-Ala-Val-Pro-Ala-Val-Cys-Val-Pro (1.59 kDa), exhibited higher protective activity against polyunsaturated fatty acid (PUFA) peroxidation than the native antioxidants, ascorbic acid and alpha-tocopherol. In a free radical scavenging assay using electron spin resonance spectroscopy, hydroxyl radical formation was quenched by 75.04% in the presence of M. coruscus peptide (50 microg/mL), which was similar to ascorbic acid. In addition, the purified peptide could also quench super-oxide and carbon-centered radicals, but those activities were weaker than for ascorbic acid. This study showed that the low-molecular-weight peptide released from in vitro gastrointestinal digestion of mussel exhibited potent antioxidant potential by inhibiting the formation of reactive oxygen species formed by the peroxidation of PUFAs.

Inhibitory effects of Orostachys japonicus extracts on the formation of N-nitrosodimethylamine.

In Korea, Orostachys japonicus has been used traditionally as a drug and health food. The aim of this study was to investigate possible inhibitory effects of O. japonicus extracts on the formation of N-nitrosodimethylamines (NDMA). Chloroform extraction was the most effective method for recovering the highest number of phenolic compounds and flavonoids; in these extracts the greatest nitrite-scavenging activity and inhibition of NDMA formation occurred at pH 2.5. The chloroform extract was separated into 10 fractions (J1-J10); fraction J4 inhibited NDMA formation by 90.1 +/- 0.4%. This fraction was then separated into five subfractions (J4-1-J4-5) using a silica gel column. Subfractions J4-2 [(+)-catechin] and J4-4 (3,4-dihdroxybenzoic acid) inhibited NDMA formation by 89.5 +/- 0.9 and 77.6 +/- 0.8%, respectively.

Inhibition of protein tyrosine phosphatase 1B by lignans from Myristica fragrans.

Inhibition of protein tyrosine phosphatase 1B (PTP1B) has been proposed as one of the drug targets for treating type 2 diabetes and obesity. Bioassay-guided fractionation of a MeOH extract of the semen of Myristica fragrans Houtt. (Myristicaceae) afforded PTP1B inhibitory compounds, meso-dihydroguaiaretic acid (1) and otobaphenol (2). Compounds 1 and 2 inhibited PTP1B with IC(50) values of 19.6 +/- 0.3 and 48.9 +/- 0.5 microM, respectively, in the manner of non-competitive inhibitors. Treatment with compound 1 on 32D cells overexpressing the insulin receptor (IR) resulted in a dose-dependent increase in the tyrosine phosphorylation of IR. These results indicate that compound 1 can act as an enhancing agent in intracellular insulin signaling, possibly through the inhibition of PTP1B activity.

Antimutagenic, antioxidant and free radical scavenging activity of ethyl acetate extracts from white, yellow and red onions.

The beneficial effects of red, yellow and white onion extracts have been assessed by antioxidant activity and antimutagenic activity. And the effects compared to BHT and ascorbic acid. Total phenolic compounds and flavonoids in onion extracts were determined. Yellow onion extract had more organic acid and free sugar than those detected in the white and red onion extract. The scavenging activity of DPPH radical and H(2)O(2) were increased depending on the concentration. The antioxidant activities using beta-carotene-linoleate system and reducing power were increased but the effect was small to that of BHT and ascorbic acid. After digested, extracts showed antimutagenic activities, and it seems that they inhibit the mutagenicity for digesting. This study demonstrated that the antimutagenicities and antioxidant properties of ethyl acetate extract against mutagens were related to their phenols and flavonoids, which are heat stable and losses digestive juices are relatively low.

Inhibitory effect of whole strawberries, garlic juice or kale juice on endogenous formation of N-nitrosodimethylamine in humans.

In vitro and in vivo experiments were performed on inhibition of nitrosation by strawberry, garlic, and kale extracts. Strawberry, garlic, and kale extracts inhibited nitrosation in vitro. However, garlic extract has a greater ability to inhibit the chemical nitrosation in vitro than strawberry and kale extracts. The garlic methanol-soluble fraction of the garlic extract was fractionated into G1-G4 fractions by Prep-LC. Fraction G1 inhibited N-nitrosodimethylamine (NDMA) formation by 84+/-1%. We studied the formation of the carcinogen NDMA in humans after administration of nitrate (400 mg/day) in combination with an amine-rich diet and its possible inhibition by administration of whole strawberries (300 g), garlic juice (200 g: 75 g garlic juice in drinking water), or kale juice (200 g) in 27 males and 13 females (ten healthy volunteers in each group) of age 24+/-3 years. Nitrate intake resulted in a significant rise in mean salivary nitrate and nitrite concentrations. Also, nitrate excretion in urine during the experimental day was significantly increased compared with the control days. When whole strawberries, garlic juice, or kale juice was provided immediately after an amine-rich diet with a nitrate, NDMA excretion was decreased by 70, 71, and 44%, respectively, compared with NDMA excretion after ingestion of an amine-rich diet with a nitrate. These results suggest that consumption of whole strawberries, garlic juice, or kale juice can reduce endogenous NDMA formation.

Effects of extract from Angelica keiskei and its component, cynaroside, on the hepatic bromobenzene-metabolizing enzyme system in rats.

The effects of Angelica keiskei Koidz. on hepatic lipid peroxide and the activities of free radical generating and scavenging enzymes were investigated in bromobenzene-induced hepatic lipid peroxidation in rats. The level of lipid peroxide elevated by bromobenzene was significantly reduced by the methanol extract from the aerial parts of A. keiskei and its component, cynaroside. Epoxide hydrolase activity was decreased significantly by the treatment of bromobenzene. However, the enzyme activity was restored in the liver of rats given the methanol extract and cynaroside. The results suggest that the reduction of bromobenzene-induced hepatic lipid peroxidation by the extract of A. keiskei and cynaroside under our experimental conditions is thought to be through enhancing the activity of epoxide hydrolase, an enzyme removing bromobenzene epoxide.