The Effects of 1-O-Acetylbritannilactone Isolated from Inula britannica Flowers on Human Neutrophil Elastase and Inflammation of RAW 264.7 Cells and Zebrafish Larvae.

During a search for natural inflammatory inhibitors, 1-O-acetylbritannilactone (ABL), a sesquiterpene lactone, was isolated from the flowers of Inula britannica. ABL significantly inhibited human neutrophil elastase (HNE) with a half-maximal inhibitory concentration (IC50) of 3.2 ± 0.3 µM, thus did so more effectively than the positive control material (epigallocatechin gallate) (IC50 7.2 ± 0.5 µM). An enzyme kinetic study was performed. ABL noncompetitively inhibited HNE with an inhibition constant Ki of 2.4 µM. ABL inhibited lipopolysaccharide-induced nitric oxide and prostaglandin E2 production by RAW 264.7 cells in a dose-dependent manner, as well as the protein-level expression of inducible nitric oxide synthase and cyclooxygenase-2. The anti-inflammatory effect of ABL was confirmed using a transgenic Tg(mpx:EGFP) zebrafish larval model. The exposure of the larvae to ABL inhibited neutrophil recruitment to the site of injury after tail fin amputation.

A novel anti-melanogenic agent, KDZ-001, inhibits tyrosinase enzymatic activity.

BACKGROUND: The demand for anti-melanogenic agents is increasing due to the unwanted side effects of current treatments. To find an effective anti-melanogenic agent, we used zebrafish as a whole animal model for phenotype-based drug and cosmetic discovery screening. OBJECTIVES: The aim of this study was to identify and explore a small molecule that could be used for skin-whitening cosmetics. METHODS: Using zebrafish embryos, we examined the effects of 1000 compounds on zebrafish development and pigmentation. Pigmentation production was assessed by tyrosinase (TYR) enzymatic activity and melanin contents. Pigmentation marker expression in the human melanoma cell line HMV-II was analyzed by western blot. We also tested reconstituted human skin tissue and analyzed KDZ-001 with computational molecular modeling. RESULTS: We identified three compounds that affected the pigmentation of developing melanophores in zebrafish. Among them, we identified KDZ-001, a novel anti-melanogenic agent, which strongly inhibits melanin synthesis in the developing melanophores of zebrafish, HMV-II cells, and reconstituted human skin with no toxicity. We found that KDZ-001 directly inhibits TYR enzymatic activity. Notably, computational molecular modeling of KDZ-001 suggested that its interaction with copper ions in the active site of TYR is essential for melanin synthesis, further demonstrating that KDZ-001 mainly acts as a TYR inhibitor to synthesize melanin. CONCLUSION: KDZ-001 inhibits melanin synthesis and has a potential for use in skin-whitening cosmetics.

Effectiveness of Ginger Essential Oil on Postoperative Nausea and Vomiting in Abdominal Surgery Patients.

BACKGROUND: The purpose of this study was to examine the effectiveness of aromatherapy with ginger essential oil on nausea and vomiting in abdominal surgery patients. METHOD: This was a quasi-experimental study with a nonequivalent control group and repeated measures. The experimental group (n = 30) received ginger essential oil inhalation. The placebo control group (n = 30) received normal saline inhalation. The level of postoperative nausea and vomiting was measured using a Korean version of the Index of Nausea, Vomiting, and Retching (INVR) at baseline and at 6, 12, and 24 h after aromatherapy administration. The data were collected from July 23 to August 22, 2012. RESULTS: Nausea and vomiting scores were significantly lower in the experimental group with ginger essential oil inhalation than those in the placebo control group with normal saline. In the experimental group, the nausea and vomiting scores decreased considerably in the first 6 h after inhaled aromatherapy with ginger essential oil. CONCLUSION: Findings indicate that ginger essential oil inhalation has implications for alleviating postoperative nausea and vomiting in abdominal surgery patients.

Cnidium officinale extract and butylidenephthalide inhibits retinal neovascularization in vitro and in vivo.

BACKGROUND: Retinal neovascularization, which is the pathological growth of new blood vessels, is associated with retinopathy of prematurity, neovascular age-related macular degeneration, diabetic retinopathy and retinal vein occlusion. In this study, we evaluated the effect of an extract of Cnidium officinale Makino (COE) and its bioactive compound, butylidenephthalide (BP), on the migration and tube formation of human umbilical vein endothelial cells (HUVECs), and on retinal pathogenic neovascularization in the oxygen-induced retinopathy (OIR) mouse model. METHOD: The HUVECs were incubated with COE and BP (0.1-10 µg/ml). The mice were exposed to 75 % oxygen for 5 days starting on the 7(th) postnatal day (P7-P12). Then, the mice were returned to room air and intraperitoneally injected with COE (100 mg/kg) and BP (5 mg/kg) once per day for 5 days (P12-P16). On P17, we measured retinal neovascularization and analyzed the angiogenesis-related proteins expression using protein arrays. RESULTS: COE and BP inhibit the HUVECs migration and the tube formation in a dose-dependent manner. In addition, COE significantly decreased retinal neovascularization in the OIR mice. COE reduced the expression levels of AREG, ANG, DLL4, Endostatin, IGFBP-2 and VEGF. Additionally, BP also inhibited the retinal neovascularization and down-regulated the expression of AREG, ANG, DLL4 and VEGF. CONCLUSION: These results suggest that COE and BP exerts antiangiogenic effects on retinal neovascularization by inhibiting the expression of AREG, ANG, DLL4 and VEGF, indicating that antiangiogenic activities of COE may be in part due to its bioactive compound, BP.

Effect of Guibi-Tang, a Traditional Herbal Formula, on Retinal Neovascularization in a Mouse Model of Proliferative Retinopathy.

Ocular pathologic angiogenesis is an important causative risk factor of blindness in retinopathy of prematurity, proliferative diabetic retinopathy, and neovascular macular degeneration. Guibi-tang (GBT) is a frequently used oriental herbal formula in East Asian countries, and is also called Qui-pi-tang in Chinese and Kihi-To in Japanese. In the present study, we investigated the preventive effect of GBT on retinal pathogenic neovascularization in a mouse model of oxygen-induced retinopathy (OIR). C57BL/6 mice were exposed to 75% hyperoxia for five days on postnatal day 7 (P7). The mice were then exposed to room air from P12 to P17 to induce ischemic proliferative retinopathy. GBT (50 or 100 mg/kg/day) was intraperitoneally administered daily for five days (from P12 to P16). On P17, Retinal neovascularization was measured on P17, and the expression levels of 55 angiogenesis-related factors were analyzed using protein arrays. GBT significantly decreased retinal pathogenic angiogenesis in OIR mice, and protein arrays revealed that GBT decreased PAI-1 protein expression levels. Quantitative real-time PCR revealed that GBT reduced vascular endothelial growth factor (VEGF), fibroblast growth factor 2 (FGF2), and plasminogen activator inhibitor 1 (PAI-1) mRNA levels in OIR mice. GBT promotes potent inhibitory activity for retinal neovascularization by decreasing VEGF, FGF2, and PAI-1 levels.

Cinidium officinale and its Bioactive Compound, Butylidenephthalide, Inhibit Laser-Induced Choroidal Neovascularization in a Rat Model.

Choroidal neovascularization (CNV) is a common pathology in age-related macular degeneration. In this study, we evaluated in a rat model the effect of an extract of Cinidium officinale Makino and its bioactive compound, butylidenephthalide, on laser-induced CNV. Experimental CNV was induced in Long-Evans rats by laser photocoagulation. C. officinale extract (COE) and butylidenephthalide was intraperitoneally injected once per day for ten days after laser photocoagulation. Choroidal flat mounts were prepared to measure CNV areas and macrophage infiltration. We used a protein array to evaluate the expression levels of angiogenic factors. The CNV area and macrophage infiltration in COE-treated rats were significantly lower than in vehicle-treated rats. COE decreased the expression levels of IGFBP-1, MCP-1, PAI-1, and VEGF. Additionally, butylidenephthalide also inhibited the laser-induced CNV formation and macrophage infiltration and down-regulated the expression of IGFBP-1, MCP-1 and VEGF. These results suggest that COE exerts anti-angiogenic effects on laser-induced CNV by inhibiting the expression of IGFBP-1, MCP-1, and VEGF, indicating that anti-angiogenic activities of COE may be in part due to its bioactive compound, butylidenephthalide.

Antibacterial effect of grapefruit seed extract (GSE) on Makgeolli-brewing microorganisms and its application in the preservation of fresh Makgeolli.

UNLABELLED: To develop a new preservation method, the antimicrobial activity of grapefruit seed extract (GSE) against Makgeolli-brewing microorganisms and food-borne pathogens was assessed, and a general analysis and sensory evaluation of fresh Makgeolli with added GSE was made. The minimum inhibitory concentration (MIC) values of GSE against 10 strains of Makgeolli-brewing microorganism were 0.0122 to 1.5625 µL/mL. The MIC values against 6 strains of food-borne pathogens were 0.0061 to 0.7813 µL/mL. On addition of 0.1% (v/v) and 0.2% GSE in bottled fresh Makgeolli, no significant difference in the pH, or the contents of total acids, ethanol, or methanol in the Makgeolli, were observed compared with control Makgeolli (with no GSE), during the preservation period (8 weeks) at 10 °C. In the Makgeolli with 0.1% and 0.2% GSE, the total bacterial counts decreased significantly by 4.9% (P < 0.01) and 11.2% (P < 0.001), respectively, versus the control. The decreases in yeast count were significantly lessened by 15.33% and 15.24% (both P < 0.001), respectively, after 8 weeks of storage, compared with the control. In the sensory evaluation of Makgeolli with 0.1% and 0.2% GSE, the refreshment and overall acceptability received significantly better scores than the control (P < 0.01), with no change in sweetness, bitterness, sourness, turbidity, color, or odor. These results suggest that GSE controls the growth of Makgeolli-brewing microorganisms and extends the shelf life (ca. 2 wk), without decreasing overall acceptance. PRACTICAL APPLICATION: A new preservation method for fresh Makgeolli by adding grapefruit seed extract (GSE) was developed. As fresh Makgeolli contains live microorganisms, the preservation period is 1 wk, which is relatively short. GSE controls the growth of Makgeolli-brewing and Makgeolli-spoiling microorganisms. 0.1% to 0.2% GSE is optimum for prolonging the shelf life (2 wk) of bottled fresh Makgeolli, and has no adverse effect on overall acceptability. We demonstrated that GSE is an effective natural additive that prolongs the shelf life of fresh Makgeolli with no significant loss in quality.

Caffeoylated phenylpropanoid glycosides from Brandisia hancei inhibit advanced glycation end product formation and aldose reductase in vitro and vessel dilation in larval zebrafish in vivo.

In our continuing efforts to identify effective naturally sourced agents for diabetic complications, five caffeoylated phenylpropanoid glycosides, acteoside (1), isoacteoside (2), poliumoside (3), brandioside (4), and pheliposide (5) were isolated from the 80% EtOH extract of Brandisia hancei stems and leaves. These isolates (1-5) were subjected to an in vitro bioassay evaluating their inhibitory activity on advanced glycation end product formation and rat lens aldose reductase activity. All tested compounds exhibited significant inhibition of advanced glycation end product formation with IC50 values of 4.6-25.7 µM, compared with those of aminoguanidine (IC50=1,056 µM) and quercetin (IC50=28.4 µM) as positive controls. In the rat lens aldose reductase assay, acteoside, isoacteoside, and poliumoside exhibited greater inhibitory effects on rat lens aldose reductase with IC50 values of 0.83, 0.83, and 0.85 µM, respectively, than those of the positive controls, 3,3-tetramethyleneglutaric acid (IC50=4.03 µM) and quercetin (IC50=7.2 µM). In addition, the effect of acteoside on the dilation of hyaloid-retinal vessels induced by high glucose in larval zebrafish was investigated. Acteoside reduced the diameters of high glucose-induced hyaloid-retinal vessels by 69% at 10 µM and 81% at 20 µM, compared to the high glucose-treated control group. These results suggest that B. hancei and its active components might be beneficial in the treatment and prevention of diabetic vascular complications.

Proanthocyanidins from Spenceria ramalana and their effects on AGE formation in vitro and hyaloid-retinal vessel dilation in larval zebrafish in vivo.

Three new A-type proanthocyanidins (1-3), ent-epiafzelechin-(2α→O→7,4α→8)-ent-afzelechin 3'-O-ß-D-glycopyranoside (1), ent-epiafzelechin-(2α→O→7,4α→8)-ent-epiafzelechin-(2α→O→7,4α→8)-ent-afzelechin (2), and ent-epiafzelechin-(2α→O→7,4α→8)-ent-epicatechin-(2α→O→7,4α→8)-ent-afzelechin (3), and three known compounds (4-6) were isolated from the whole plant of Spenceria ramalana. The structures of the new proanthocyanidins were established by spectroscopic and chemical studies. The inhibitory effects of compounds 1-6 on the formation of advanced glycation end products were examined in vitro. Compounds 3 and 6 showed the strongest inhibition, with IC50 values of 17.4 ± 0.5 and 14.1 ± 1.6 µM, respectively. The effects of these isolates on the dilation of hyaloid-retinal vessels induced by high glucose (HG) in larval zebrafish were also investigated. Compound 3 reduced the dilation of HG-induced hyaloid-retinal vessels most effectively. This compound reduced the diameters of HG-induced hyaloid-retinal vessels by about 157.7% and 164.1% at 10 and 20 µM, respectively, versus the HG-treated control group.

Examination of 1-methylimidazole series ionic liquids in the extraction of flavonoids from Chamaecyparis obtuse leaves using a response surface methodology.

Ionic liquids (ILs) are a new type of reagent that has accelerated research in extraction technology. On the other hand, few studies have systematically applied 1-methylimidazole ([MIM]) series ILs to the extraction of bioactive compounds from plants. In this study, [MIM] series ILs were used to extract four bioactive flavonoids, such as dihydrokaempferol, quercitrin, amentoflavone and myricetin, from Chamaecyparis obtuse (CO) leaves. First, a screen of the extraction method and solvent revealed the [MIM] series ILs to be suitable as additives in methanol in Soxhlet extraction. Second, an examination of a range of cations and anions of [MIM] series ILs for extraction revealed 1-decyl-3-methylimidazolium bromide ([DMIM][Br]) to be the best selection as an additive in methanol for the Soxhlet extraction of flavonoids from (CO) leaves. Finally, some factors of extraction, such as temperature, time and amount of samples, were examined systematically using a response surface methodology (RSM). Based on the above optimization, 2.41, 3.47, 0.76 and 3.15mg/g of dihydrokaempferol, quercitrin, amentoflavone and myricetin, respectively, were extracted from 15g of CO leaves by 2.5mgmL(-1) of [DMIM][Br] as additives in 200mL of methanol in Soxhlet extraction at 200°C for 8h. This study highlights the potential of [MIM] series ILs as promising reagents for the extraction of bioactive compounds from plants.