Enhancing Effect of Trachelogenin from Trachelospermi caulis Extract on Intestinal Barrier Function.

Trachelospermi caulis is used widely as an herbal medicine in oriental countries to attenuate fever and pain. We wished to reveal the novel function of this herb and its active component on barrier function in intestinal epithelial cells. Monolayers of intestinal epithelial cells (Caco-2) were used to evaluate the transepithelial electrical resistance (TEER) and quantity of permeated ovalbumin (OVA) as indices of barrier function. T. caulis increased TEER values on cell monolayers and decreased OVA permeation across cell monolayers. To ascertain the active component of T. caulis, the extract was isolated to five fractions, and the effect of each of these fractions on intestinal barrier function examined. Chloroform and ethyl acetate fractions showed increased TEER values and decreased OVA flux. Chloroform and ethyl acetate fractions contained mainly trachelogenin and its glycoside, tracheloside. Trachelogenin increased TEER values and decreased OVA flux by enhancing the tight-junction protein occludin (but not tracheloside) in Caco-2 monolayers. These findings demonstrated that trachelogenin, an active component of T. caulis, might help to attenuate food allergy or inflammatory bowel disease through inhibition of allergen permeation or enhancement of the intestinal barrier.

Bioavailability of ginsenosides from white and red ginsengs in the simulated digestion model.

This study aims to investigate the bioavailability of ginsenosides during simulated digestion of white (WG) and red (RG) ginseng powders. Stability, bioaccessibility, and permeability of ginsenosides present in WG and RG were studied in a Caco-2 cell culture model coupled with oral, gastric, and small intestinal simulated digestion. Most ginsenosides in WG and RG were stable (>90%) during the simulated digestion. Bioaccessibilities of total ginsenosides during in vitro digestion of WG and RG were similar at approximately 85%. However, the bioaccessibility of protopanaxatriol type ginsenosides in the early food phase was greater than that of the protopanaxadiol type. The less polar RG ginsenosides were released later following the jejunum phase. Ginsenosides had low permeability (<1 × 10(-6) cm/s) through Caco-2 cell monolayers. These findings suggest that the WG and RG ginsenoside compositions affect bioaccessibility during digestion and that ginsenosides are poorly absorbed in humans.

Diferuloylputrescine, a predominant phenolic amide in corn bran, potently induces apoptosis in human leukemia U937 cells.

The purpose of this study was to investigate how proliferation and apoptosis in human leukemia U937 cells are affected by four hydroxycinnamic acid derivatives (HCADs) in corn (Zea mays L.) bran: p-coumaric (CA), ferulic acids (FA), dicoumaroylputrescine (DCP), and diferuloylputrescine (DFP). Of the four HCADs, DFP dose dependently exerted the strongest cytotoxic effect and induction of apoptosis in the U937 cells. In addition, DFP induced distinct morphological changes characteristic of cellular apoptosis, such as chromatin condensation, apoptotic bodies, and DNA fragmentations. The DFP-induced apoptosis was also associated with released cytochrome c in the cytosol with activation of caspase 3, together with the downregulation of anti-apoptotic proteins, including XIAP and cIAP2, Bcl-2, and Mcl-1. Finally, the DFP-induced apoptosis was a cell-specific response in leukemia cells, as compared with those of other cancer cells, such as Caki, HT29, SK-Hep1, and MDA-MB231. Thus, these results suggest that DFP may be useful as a potential source of natural antileukemic agents.

Anti-inflammatory and anti-osteoarthritis effects of fermented Achyranthes japonica Nakai.

ETHNOPHARMACOLOGICAL RELEVANCE: Achyranthyes japonica Nakai (AJN) has been traditionally used to control pain and improve dysfunction in osteoarthritis (OA) patients. AIM OF THE STUDY: The objectives of the present study were to investigate anti-inflammatory and anti-osteoarthritis activities of fermented AJN (FAJN). MATERIALS AND METHODS: Anti-inflammatory activity of non-fermented AJN (NFAJN) and FAJN was evaluated by in vitro assay using LPS-induced RAW 264.7 cells. In addition, their cartilage protective effects were also determined in vitro assay using SW1353 cell and in vivo model system using collagenase-induced arthritis (CIA) in rabbits. Moreover, we isolated and identified 20-hydroxyecdysone (20-HES) as a marker component in FAJN. RESULT: FAJN showed stronger anti-inflammatory activity than NFAJN through inhibiting production of NO and PGE2 in LPS-induced RAW 264.7, and lowering levels of MMP-3 release in SW1353 cells treated with TNF-a. FAJN contained higher levels of 20-HES, as a marker component, than AJN. FAJN ameliorates the progress of OA by inhibiting local inflammation. It does this by regulating levels of TNF-a and IL-4, and protecting articular cartilage by preventing destruction of proteoglycan, collagens, and also preventing injury to chondrocytes. CONCLUSION: Therefore, FAJN is a potential therapeutic agent for reduction of cartilage damage that occurs in OA.

Anti-inflammatory activity of hydroxycinnamic acid derivatives isolated from corn bran in lipopolysaccharide-stimulated Raw 264.7 macrophages.

In this study, the effect of the 80% ethanolic extract of corn bran (EECB) on inhibition of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells was investigated. The EECB inhibited LPS-induced NO production and iNOS expression in a dose-dependent manner. Four hydroxycinnamic acid derivatives (HADs), including two free cinnamic acids, p-coumaric acid (CA) and ferulic acid (FA), and their conjugate phenolic amides, p-dicoumaroyl-putrescine (DCP) and diferuloylputrescine (DFP), were found to be present in the EECB by LC-MS analysis, and DFP (378.66 µg/g) was the predominant phenolic compound, followed by DCP (7.83 µg/g)>CA (5.58 µg/g)>FA (1.84 µg/g). The four HADs significantly inhibited NO production and iNOS expression in a dose-dependent manner. Among the four HADs tested, DFP showed the most potent inhibition on NO production and iNOS mRNA and protein expression, followed by DCP>FA ≥ CA. DFP also exhibited the strongest inhibition on LPS-induced iNOS and NF-κB luciferase activity, which was followed by DCP ≥ FA (CA)>CA (FA). Thus, these results suggest that phenolic amides in the corn bran may be a potential source of natural anti-inflammatory agents.

Effects of a safflower tea supplement on antioxidative status and bone markers in postmenopausal women.

We conducted this study to examine the effects of safflower seed granular tea containing physiologically active polyphenols on antioxidative activities and bone metabolism. Forty postmenopausal women ages 49 to 64-years were recruited from Daegu and Gyeongbuk and were randomly assigned to either a safflower tea supplement (Saf-tea) group (n = 27) or a placebo group (n = 13). The Saf-tea group received 20 g of safflower seed granule tea per day containing a 13% ethanol extract of defatted safflower seeds, whereas the placebo group received a similar type of tea that lacked the ethanol extract. No significant changes in nutrient intake for either the placebo or Saf-tea groups were observed before or after the study period, except vitamin A intake increased after 6 months in the Saf-tea group. Dietary phytoestrogen intakes were similar in the Saf-tea group (60.3 mg) and placebo group (52.5 mg). Significant increases in plasma genistein and enterolactone were observed in the Saf-tea group. After 6 months of supplementation, serum levels of antioxidant vitamins such as α-tocopherol and ascorbic acid increased significantly, and TBARS levels decreased in the Saf-tea group compared to the placebo group. Serum osteocalcin levels were reduced (P < 0.05) in the Saf-tea group after 6 months, whereas serum osteocalcin did not change in the placebo group. Urinary deoxypyridinoline/creatinine excretion was not different between the two groups at baseline, and did not change in either group after 6 months. Bone mineral density decreased significantly in the placebo group (P < 0.01) but not in the supplemented group. It was concluded that polyphenols (72 mg/day), including serotonin derivatives, in the Saf-tea had both antioxidant and potential bone protecting effects in postmenopausal women without liver toxicity.

Enrichment of pinolenic acid at the sn-2 position of triacylglycerol molecules through lipase-catalyzed reaction.

Reports have shown that Delta-5 polyunsaturated fatty acids (-5 PUFA) are enriched at sn-1,3 positions of triacylglycerols (TAG) in pine (Pinus koraiensis) nut oil (Pn). As a major Delta-5 PUFA, pinolenic acid (Pi) is about 14.2% in the oil, while the percentage of Pi at the sn-1 and/or sn-3 positions in TAG was found more than 20%. In this current study, the enhancement of Pi at the sn-2 position has been achieved by acyl migration during the lipase-catalyzed inter-esterification between Pn and palm stearin (Ps). After reaction, the proportion of Pi increased at sn-2 positional fatty acid even is similar to that in total fatty acid; for example, in the inter-esterified product of 50:50 (Pn:Ps), the same amount of Pi (7.1%) present was detected both at the sn-2 and sn-1,3 positions. However, the reduction of phytosterols and tocopherols are observed in the inter-esterified products.

Antioxidant and antithrombus activities of enzyme-treated Salicornia herbacea extracts.

This study was attempted to investigate antioxidant and antithrombus activities of water and methanol extracts of enzyme-treated Salicornia herbacea (SH)by in vitro assays observing the inhibitory activity of a rat liver microsomal lipid peroxidation, DPPH radical scavenging activity, activated partial thromboplastin times (APTT) and thromboplastin times (TT). The water and methanol extracts from enzyme-treated SH inhibited the lipid peroxidation in a dose-dependent manner over a concentration range of 0.1-1.0 mg/ml. The activity of enzyme-treated water and methanol extracts was stronger than that of non-enzyme-treated water and methanol extracts. The inhibitory activity of the water extract was higher at a concentration of 1.0 mg/ml than that of the methanol extract. The activity was the highest in the enzyme-treated water extract, and was approximately 1.08 times higher than alpha-tocopherol, a natural antioxidant. The DPPH radical scavenging activities of the SH extracts were similar to their lipid peroxidation inhibitory activity. The APTT of the water and methanol extracts was delayed at a concentration range of 0.25-2.0 mg/ml in a dose-dependent manner. The APTT of the methanol extract was longer at a concentration of 1.0 mg/ml than that of the water extracts. The enzyme-treated methanol extract exhibited the longest APTT even at a concentration of 0.50 mg/ml. The TT activities of the SH extracts were also similar to their APTT activities. These results suggest that water and methanol extracts of the enzyme-treated SH may be useful as potential antioxidant and antithrombus sources, respectively.

Effects of defatted safflower seed extract and phenolic compounds in diet on plasma and liver lipid in ovariectomized rats fed high-cholesterol diets.

Six polyphenolic compounds were isolated from ethylacetate extract secondary to 80% ethanol extraction of defatted safflower seeds. They were categorized into three types: lignans, flavones and serotonin derivatives. Female Sprague-Dawley rats weighing 163.4 +/- 6.3 g were ovariectomized (Ovx) and fed either ethylacetate extract at a level of 1% (w/w) or three types of safflower polyphenolic compounds at a level of 200 mg/kg in a diet containing 0.5% (w/w) cholesterol for four wk. The sham and Ovx control groups were fed the same diet without safflower components. Plasma GOT and GPT levels did not differ among the six experimental groups. The plasma levels of total cholesterol were reduced in the four safflower groups by 20-30% as compared to the Ovx control. The plasma level of HDL-cholesterol was higher in the Ovx+ethylacetate extract group or appeared to be in the three Ovx+safflower polyphenolic groups than in the Ovx control. The level of plasma triglyceride was also significantly lower in the Ovx+lignan group than in the Ovx control. The liver level of cholesterol was significantly reduced in the Ovx+ethylacetate extract group. Fecal excretion of cholesterol increased by the safflower lignans and flavones, whereas that of bile acid was not significantly changed by the safflower polyphenols. Matairesinol and acacetin isolated from safflower seeds reduced the cholesterol content in cultured HepG2 cells at a concentration of 0.01-0.1 microM and all three safflower polyphenolics decreased triglyceride content at the concentration of 0.1 microM. These results suggest that safflower polyphenols have the effect of improving blood lipid status via increasing HDL-cholesterol formation and cholesterol excretion without significant uterotropic action in estrogen-deficient animals.

Effects of forsythia fruit extracts and lignan on lipid metabolism.

A primary methanol extract (F-ME), secondary butanol-soluble fraction (F-BU), and lignans were prepared from forsythia fruit (Forsythia viridissima L.) and added to 0.5% (w/w) cholesterol diets for male Sprague-Dawley rats weighing 121 +/- 12 g. There were six experimental groups: a control group, 0.2%, 0.4% F-ME supplemented groups, 0.1%, 0.02% F-BU groups and 0.02% lignan group. After 3 weeks of feeding, body weight gains, serum GOT and GPT levels were not different among the groups. HDL-/total cholesterol ratios increased in the 0.2% F-BU and lignan groups compared with the control groups. Liver triglyceride level lowered in most of forsythia groups. Fecal cholesterol excretions increased in the lignan group. Arctiin isolated from the forsythia fruit reduced cholesterol and triglyceride contents in cultured HepG2 cells at 0.01-0.1 microM. These results indicated that the forsythia lignan, arctiin is effective on improving blood lipid status without a significant hepatotoxicity and is to be utilized for the functional foods for lipid-lowering action.

Proliferative effects of flavan-3-ols and propelargonidins from rhizomes of Drynaria fortunei on MCF-7 and osteoblastic cells.

The proliferative effects of thirty Oriental medicinal herbs on MCF-7 (estrogen-sensitive breast cancer cell line) and ROS 17/2.8 osteoblast-like cells were determined using the MTT assay. Methanol extracts from several herbs was found to show proliferative activity on the above two cell lines in the range of 5 to 100 microg/mL. Among these active herbs, the methanol extract from the rhizomes of Drynaria fortunei showed the most potent proliferative activity, and the cell proliferations were significantly increase by 136 and 158% in the MCF-7 and ROS 17/2.8 cells, respectively, when treated with 100 microg/mL. Through a bioassay-guided separation, eight flavonoids, including four new flavan-3-ols and two propelargonidins, together with the known (-)-epiafzelechin and naringin, were isolated. Their chemical structures were characterized as (-)-epiafzelechin (1), (-)-epiafzelechin-3-O-beta-D-allopyranoside (2), (-)-epiafzelechin-3-O-(6"-O-acetyl)-beta-D-allopyranoside (3), 4beta-carboxymethyl-(-)-epiafzelechin methyl ester (4), 4beta-carboxymethyl-(-)-epiafzelechin sodium salt (5), naringin (6), (-)-epiafzelechin-(4beta-->8)-4beta-carboxymethylepiafzelechin methyl ester (7) and (-)epiafzelechin-(4beta-->8, 2beta-->O-->7)-epiafzelechin-(4beta-->8)-epiafzelechin (8) by extensive 1D and 2D NMR spectroscopy. Most of these flavonoids, in the range of 10(-15) to approximately 10(-6) M, accelerated the proliferation of MCF-7 cell, with compounds 7 and 8, in the range of 10(-15) to approximately 10(-12) M, showing especially potent proliferation effects. Meanwhile, seven flavonoids, with the exception of compound 4, stimulated the proliferation of ROS 17/2.8 cells in the range of 10(-15) to approximately 10(-6) M, with compounds 5-8 especially accelerating the proliferation, in dose-dependent manners (10(-15) to approximately 10(-9) M), and their proliferative effect was much stronger than that of E2 and genistein. These results suggest that propelargonidin dimers and trimers isolated from the rhizomes of Drynaria fortunei may be useful as potential phytoestrogens, which play important physiological roles in the prevention of postmenopausal osteoporosis.