Pigments in green tea leaves (Camellia sinensis) suppress transformation of the aryl hydrocarbon receptor induced by dioxin

Environmental contaminants such as dioxins enter the body mainly through diet and cause various toxicities through transformation of the aryl hydrocarbon receptor (AhR). We previously reported that certain natural flavonoids at the dietary level suppress the AhR transformation induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In this study, we identified lutein and chlorophyll a and b from green tea leaves as the novel antagonists for AhR. These active compounds suppressed AhR transformation dose-dependently with the 50% inhibitory concentration (IC(50)) values against 0.1 nM TCDD-induced AhR transformation at 3.2, 5.0, and 5.9 microM, respectively. (-)-Epigallocatechin gallate, which is the most abundant flavonoid in green tea leaves, also showed stronger suppressive effects than did other major tea components, with the IC(50) value of 1.7 microM. Thus, these pigments of green tea leaves have the potential to protect from dioxin toxicity through the suppression of AhR transformation.     

Suppressive effects of ethanolic extracts from propolis and its main botanical origin on dioxin toxicity

Suppressive effects of ethanolic extracts prepared from propolis group 12 and its main botanical origin (leaf bud of Baccharis dracunculifolia) on transformation of the aryl hydrocarbon receptor (AhR), the initial action of dioxin toxicity, were investigated. It was found that suppressive effects of propolis on AhR transformation were relatively higher than those of resins of its botanical origin in cell-free system and in Hepa-1c1c7 cells. When the composition of chemical ingredients was measured, propolis contained slightly higher amounts of flavonoid aglycones as compared with its botanical origin with the same characteristics. Moreover, antiradical activity, one of the typical biological activities of flavonoids, in propolis was also slightly higher than that in its botanical origin. These results indicate that not only propolis but also its botanical origin contains high amounts of flavonoid aglycones and that both of them are useful dietary sources for flavonoids with a potency to prevent dioxin toxicity.     

Phytomonitoring and phytoremediation of agrochemicals and related compounds based on recombinant cytochrome P450s and aryl hydrocarbon receptors (AhRs)

Molecular mechanisms of metabolism and modes of actions of agrochemicals and related compounds are important for understanding selective toxicity, biodegradability, and monitoring of biological effects on nontarget organisms. It is well-known that in mammals, cytochrome P450 (P450 or CYP) monooxygenases metabolize lipophilic foreign compounds. These P450 species are inducible, and both CYP1A1 and CYP1A2 are induced by aryl hydrocarbon receptor (AhR) combined with a ligand. Gene engineering of P450 and NADPH cytochrome P450 oxidoreductase (P450 reductase) was established for bioconversion. Also, gene modification of AhRs was developed for recombinant AhR-mediated β-glucronidase (GUS) reporter assay of AhR ligands. Recombinant P450 genes were transformed into plants for phytoremediation, and recombinant AhR-mediated GUS reporter gene expression systems were each transformed into plants for phytomonitoring. Transgenic rice plants carrying CYP2B6 metabolized the herbicide metolachlor and remarkably reduced the residues in the plants and soils under paddy field conditions. Transgenic Arabidopsis plants carrying recombinant guinea pig (g) AhR-mediated GUS reporter genes detected PCB126 at the level of 10 ng/g soils in the presence of biosurfactants MEL-B. Both phytomonitoring and phytoremediation plants were each evaluated from the standpoint of practical uses.     

Antioxidant activity of vinegar produced from distilled residues of the Japanese liquor shochu

Rice shochu distilled residue (RSDR) is a byproduct of rice shochu production. RSDR was converted into vinegar by acetate fermentation. In our present study, two major antioxidant compounds, tyrosol and ferulic acid, were identified from the RSDR-derived vinegar. Furthermore, we investigated the antioxidant activity of freeze-dried RSDR-derived vinegar, which was Acetobactor aceti fermentation powder (AFP), in vitro and in vivo. AFP at 0.25 mg/mL or higher concentrations showed an inhibitory effect against lipid peroxidation and cellular GSH depletion in HepG2 cells induced by H(2)O(2) (P < 0.05). We thus considered the potential of AFP in protecting cells against damage induced by H(2)O(2). Its antioxidant activity was evaluated in vivo using carbon tetrachloride (CCl(4))-induced acute liver injury mouse models. Five consecutive days of oral preadministration of AFP dissolved in PBS at 200, 400, and 800 mg/kg body weight significantly suppressed lipid peroxidation in the liver induced by CCl(4) (P < 0.01). Consequently, treatment with AFP at 200 mg/kg body weight or higher doses suppressed the elevation of alanine aminotransferase and aspartate aminotransferase levels in serum (P < 0.05). These findings suggest that RSDR-derived vinegar can be developed as a health food with an antioxidant effect for the prevention of oxidative injury and cancer.     

Gastroprotective effect of red pigments in black chokeberry fruit (Aronia melanocarpa Elliot) on acute gastric hemorrhagic lesions in rats

It has been reported that the fruits and leaves of berries such as the blackberry, raspberry, and strawberry contain a high level of scavenging activity for chemically generated active oxygen species. This study investigated the antioxidative activities of black chokeberry fruit (Aronia melanocarpa Elliot) both in vitro and in vivo using the DPPH stable radical and rats with ethanol-induced gastric injury, respectively. The red pigment fraction of the black chokeberry contained three main components, one of which was identified as cyanidin 3-O-beta-glucoside by HPLC analysis and (1)H NMR. The black chokeberry red pigment fraction scavenged >44% of DPPH radicals at a concentration of 25 microg/mL compared to the control solution. The black chokeberry extract and its hydrolysate administrated at 2 g/kg of body weight each had nearly the same protective effect as quercetin administrated at 100 mg/kg of body weight in suppressing the area of gastric mucosal damage caused by the subsequent application of ethanol to <30% compared to the control group. The black chokeberry red pigment fraction had a similarly significant protective effect on gastric mucosa in a dose-dependent manner when administered at 30-300 mg/kg of body weight, and the administration of 30 mg/kg of body weight could suppress ethanol-induced gastric mucosal damage by approximately 50% (ID(50) = 30 mg/kg of body weight).     

Effect of garlic oil on neutrophil infiltration in the small intestine of endotoxin-injected rats and its association with levels of soluble and cellular adhesion molecules

Garlic ( Allium sativum ) possesses anti-inflammatory effects. This study investigated the effects of garlic oil on endotoxin-induced neutrophil infiltration in the small intestine. Wistar rats received by gavage 10, 50, or 100 mg/kg body wt garlic oil (GO) or the vehicle (corn oil; 2 mL/kg body wt) every other day for 2 weeks before being injected with endotoxin (ip, 5 mg/kg body wt). Control rats were administered corn oil and injected with sterile saline. Blood samples for the measurement of soluble adhesion molecules were collected at various time points after injection, and all other samples were collected 18 h after injection. The 10 and 50 mg/kg doses suppressed endotoxin-induced neutrophilia, serum levels of sL-selectin and sICAM-1, cellular CD11b on neutrophils, intestinal ICAM-1 content, and neutrophil infiltration (P < 0.05). The 100 mg/kg dose significantly lowered local ICAM-1 and cellular CD11b on neutrophils (P < 0.05) but did not have a beneficial effect on neutrophil infiltration. In addition, 100 mg/kg of GO worsened the elevation of the local TNF-α level and neutrophilia. Appropriate doses of garlic oil have a preventive effect on endotoxin-induced neutrophil infiltration and damage to the small intestine.     

Gentiana manshurica Kitagawa reverses acute alcohol-induced liver steatosis through blocking sterol regulatory element-binding protein-1 maturation

This study was undertaken to investigate the protective effects of Gentiana manshurica Kitagawa (GM) on acute alcohol-induced fatty liver. Mice were treated with ethanol (5 g/kg of body weight) by gavage every 12 h for a total of three doses to induce acute fatty liver. Methanol extract of GM (50, 100, or 200 mg/kg) or silymarin (100 mg/kg) was gavaged simultaneously with ethanol for three doses. GM administration significantly reduced the increases in serum ALT and AST levels, the serum and hepatic triglyceride levels, at 4 h after the last ethanol administration. GM was also found to prevent ethanol-induced hepatic steatosis and necrosis, as indicated by liver histopathological studies. Additionally, GM suppressed the elevation of malondialdehyde (MDA) levels, restored the glutathione (GSH) levels, and enhanced the superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities. The concurrent administration of GM efficaciously abrogated cytochrome P450 2E1 (CYP2E1) induction. Moreover, GM significantly reduced the nuclear translocation of sterol regulatory element-binding protein-1 (nSREBP-1) in ethanol-treated mice. These data indicated that GM possessed the ability to prevent ethanol-induced acute liver steatosis, possibly through blocking CYP2E1-mediated free radical scavenging effects and SREBP-1-regulated fatty acid synthesis. Especially, GM may be developed as a potential therapeutic candidate for ethanol-induced oxidative damage in liver.     

Water extract of defatted rice bran suppresses visceral fat accumulation in rats

Rice bran has been reported to inhibit pancreatic lipase activity in vitro. This action shows that administration of rice bran may result in a decrease in plasma triglyceride levels and suppress accumulation of fat in vivo. We administered water extract of defatted rice bran (WED-rice bran) to rats to determine its effects. Single administration of WED-rice bran at a dose of 1 g/kg body weight caused a decrease in plasma triglyceride levels in fat emulsion induced hypertriglyceridemic rats. Four week administration of WED-rice bran suppressed accumulation of visceral fat and body weight gain without influencing food consumption, liver function, and renal function. These results indicate that a reduction of plasma triglycerides and suppression of visceral fat accumulation may be induced by pancreatic lipase inhibition caused by administration of WED-rice bran.     

Protective effects of dietary chamomile tea on diabetic complications

Matricaria chamomilla L., known as "chamomile", has been used as an herbal tea or supplementary food all over the world. We investigated the effects of chamomile hot water extract and its major components on the prevention of hyperglycemia and the protection or improvement of diabetic complications in diabetes mellitus. Hot water extract, esculetin (3) and quercetin (7) have been found to show moderate inhibition of sucrase with IC50 values of 0.9 mg/mL and 72 and 71 microM, respectively. In a sucrose-loading test, the administration of esculetin (50 mg/kg body weight) fully suppressed hyperglycemia after 15 and 30 min, but the extract (500 mg/kg body weight) and quercetin (50 mg/kg body weight) were less effective. On the other hand, a long-term feed test (21 days) using a streptozotocin-induced rat diabetes model revealed that the same doses of extract and quercetin showed significant suppression of blood glucose levels. It was also found that these samples increased the liver glycogen levels. Moreover, chamomile extract showed potent inhibition against aldose reductase (ALR2), with an IC50 value of 16.9 microg/mL, and its components, umbelliferone (1), esculetin (3), luteolin (6), and quercetin (7), could significantly inhibit the accumulation of sorbitol in human erythrocytes. These results clearly suggested that daily consumption of chamomile tea with meals could contribute to the prevention of the progress of hyperglycemia and diabetic complications.     

Antidiabetic activity of red wine polyphenolic extract, ethanol, or both in streptozotocin-treated rats

A polyphenol extract from a Corbières (France) red wine (P, 200 mg/kg), ethanol (E, 1 mL/kg), or a combination of both (PE) was administered by daily gavage for 6 weeks to healthy control or streptozotocin (60 mg/kg i.v.)-induced diabetic rats (180-200 g). Treatment groups included C or D (untreated control or diabetic) and CP, CE, or CPE (treated control) or DP, DE, or DPE (treated diabetic). P treatment induced a reduction in body growth, food intake, and glycemia in both CP and DP groups. In DP, hyperglycemia was reduced when measured 1 h after daily treatment but not at sacrifice (no treatment on that day). The hyperglycemic response to the oral glucose tolerance test (OGTT) and plasma insulin at sacrifice were impaired similarly in DP and D groups. In contrast, in DE or DPE, body growth was partially restored while hyperglycemia was reduced both during treatment and at sacrifice. In addition, hyperglycemia response to OGTT was reduced and plasma insulin was higher in DE or DPE than in D animals, indicating a long-term correction of diabetes in ethanol-treated animals. Morphometric studies showed that ethanol partially reversed the enlarging effect of diabetes on the mesenteric arterial system while the polyphenolic treatment enhanced it in the absence of ethanol. In summary, our study shows that (i). a polyphenol extract from red wine ("used at a pharmacological" dose) reduces glycemia and decreases food intake and body growth in diabetic and nondiabetic animals and (ii). ethanol ("nutritional" dose) administered alone or in combination with polyphenols is able to correct the diabetic state. Some of the effects of polyphenols were masked by the effects of ethanol, notably in diabetic animals. Further studies will determine the effect of "nutritional" doses of polyphenols as well as their mechanism of action.     

Assessment of cytotoxicity and AhR-mediated toxicity of sediments from water level fluctuation zone in the Three Gorges Reservoir,China

Purpose

The Three Gorges Dam project is one of the biggest projects in the world. The water level fluctuation zone (WLFZ) was created with impoundment. The purpose of the current study is to investigate the ecotoxicological potential of the sediment extracts from the WLFZ and propose future WLFZ monitoring for early detection of environmental changes brought about by pollution.

Materials and methods

The investigation was performed by using cell-based in vitro bioassays to determine acute cytotoxicity (neutral red retention assay) and mechanism-specific aryl hydrocarbon receptor (AhR)-mediated activity (7-ethoxyresorufin-O-deethylase (EROD) induction assay) of sediment extracts with rainbow trout (Oncorhynchus mykiss) liver cells (RTL-W1).

Results and discussion

Results showed that the cytotoxicity and AhR-mediated toxicity potential of the sediment extracts from the WLFZ in the Three Gorges Reservoir (TGR) were moderate of level compared with the ecotoxicity of sediments from other river systems. However, according to a sediment classification system in Germany, sediments from some individual sites in the WLFZ showed strong toxicity. Compared to the results from the Yangtze River in our previous study, more attention should be paid to the aryl hydrocarbon receptor inducers in the WLFZ in TGR.

Conclusions

The in vitro bioassays used in this study may serve as a valuable tool to predict the potential ecological hazards of the organic pollutants in the WLFZ to the aquatic organisms in some extent.

     

NO合酶抑制剂 L-NAME和NO前体L-Arg对小鼠体内卵母细胞成熟的影响

摘要: 采用腹腔单独注射L-NAME(左旋硝基精氨酸甲基酯)或联合注射L-NAME和L-Arg, 取出卵母细胞体外观察，研究了NO对小鼠卵母细胞体内成熟的影响。结果表明,（1）单独注射5、10、20 mg / kg体重L-NAME均能显著抑制卵母细胞第一极体的释放(P <0.01)，尤以10 mg / kg 体重L-NAME组作用最为明显, 各剂量组对生发泡的破裂均无影响(P >0.05)；（2）10 mg / kg 体重L-NAME对卵母细胞成熟的抑制作用能被同时注射的100 mg / kg 或200 mg /kg 体重L-Arg逆转，其中以200 mg / kg体重作用最为明显;（3）L-NAME对卵母细胞畸形率的影响和培养时间有关，而对卵母细胞的存活率没有统计学影响。实验结果证明NO参与了小鼠卵母细胞的体内成熟调控。     

Relationships between shoot and root characteristics of white clover cultivars differing in response to phosphorus

Root dry weight and the relationship between shoot and root characteristics for 119 white clover (Trifolium repens L.) cultivars and breeding lines were examined at five (40, 120, 200, 400, and 500 mg P/kg soil) phosphorus (P) levels. There were significant (P<0.01) differences among cultivars for root dry weight and root/shoot ratio. Instantaneous root/shoot ratio decreased with increasing P supply up to 200 mg P/kg, and then increased again from 400 to 500 mg P/kg soil. Allometric analysis indicated that root growth relative to shoot growth increased with increasing P supply. There was a significant (P<0.01) cultivar x P level interaction such that for the cultivars, the root/shoot ratio was either unaffected, increasing, or decreasing with increasing level of P supply. However, root/shoot ratio appeared to be unrelated to the functions used to describe the P response of the 119 cultivars studied. Shoot characters such as dry weight, leaf number, and stolon length were most closely correlated with root dry weight at the moderately low P levels of 120 and 200 mg added P/kg, where average P response was 9 and 24% of yields at 500 mg P/kg soil, respectively, rather than at the higher P levels. Root/shoot ratio was significantly correlated (r>0.6) with root dry weight over all P levels but not with any of the measured shoot characters (‐0.2> r <0.3).     

In vivo antitumor effects of 4,7-dimethoxy-5-methyl-1,3-benzodioxole isolated from the fruiting body of Antrodia camphorata through activation of the p53-mediated p27/Kip1 signaling pathway

In this study, 4,7-dimethoxy-5-methyl-1,3-benzodioxole (SY-1) was isolated from three different sources of dried Antrodia camphorata (AC) fruiting bodies. AC is a medicinal mushroom that grows on the inner heartwood wall of Cinnamomum kanehirai Hay (Lauraceae), which is an endemic species that is used in Chinese medicine for its antitumor properties. We demonstrated that SY-1 [given as a 1-30 mg/kg body weight intraperitoneal (ip) injection three times per week] profoundly decreased the growth of COLO-205 human colon cancer cell tumor xenografts in an athymic nude mouse model. We further demonstrated that significant AC extract-mediated antitumor effects were observed at the highest concentration (5 g/kg body weight/day). No gross toxicity signs were observed (i.e., body weight changes, general appearance, or individual organ effects). Frozen COLO-205 xenograft tumors were pulverized in liquid N(2), and the expression of cell cycle regulatory proteins was detected by immunoblotting. We found that the p53-mediated p27/Kip1 protein was significantly induced in the low-dose (1 mg/kg body weight) SY-1-treated tumors, whereas the p21/Cip1 protein levels did not change. The G0/G1 phase cell cycle regulators induced by SY-1 were also associated with a significant decrease in cyclins D1, D3, and A. These results provide further evidence that SY-1 may have significance for cancer chemotherapy.     

土壤中Cd、P、Zn含量对水稻产量和植株中矿物浓度的影响

在温室中进行了剖析添加的Cd、P、Zn对稻谷产量、植株和稻草中的矿物含量以及糙米中Cd的浓度的研究。P增加了稻谷和植株产量,而Cd、Zn和P-Zn的交互作用使其降低。稻谷和植株产量有类似的线性回归方程式,其稻谷的线性回归方程为: Y=17.24+0.0466(P)-0.1850(Cd)-0.1115(Zn)-0.0005(P-Zn) 其R²=0.97^**,式中Y为稻谷产量(克/株);P为添加的P浓度(毫克/公斤);(Cd)为添加的Cd浓度(毫克/公斤);(Zn)为添加的Zn浓度(毫克/公斤);(P-Zn)为P和Zn的交互作用。所有处理均明显地影响着稻草中矿物元素的含量。粕米中的Cd浓度随着添加的Cd、P、Zn浓度的增加而增加;但多元回归分析表明只有Cd的影响是显著的。粕米中Cd的浓度与收获时用0.05M HCI所提取的风干土中的Cd浓度有极显著相关性(Y=0.75^**).对糙米中Cd浓度的评价进行了简要的讨论。     

Direct intestinal absorption of red fruit anthocyanins, cyanidin-3-glucoside and cyanidin-3,5-diglucoside, into rats and humans.

We determined red fruit anthocyanins, cyanidin-3-glucoside (Cy-g) and cyanidin-3,5-diglucoside (Cy-dg), incorporated into plasma and liver of rats and human plasma by UV-HPLC. Fifteen minutes after an oral supplementation of a mixture of 320 mg of Cy-g and 40 mg of Cy-dg/kg of body weight, rats showed an increase to a maximum of 1563 microg (3490 nmol) of Cy-g/L and 195 microg (320 nmol) of Cy-dg/L in plasma and 0.067 microg (0.15 nmol) of Cy-g/g and a trace of Cy-dg together with methylated metabolites such as peonidin-3-glucoside in liver. In human plasma, 30 min after intake (2.7 mg of Cy-g and 0.25 mg of Cy-dg/kg of body weight), an average of 11 microg (24 nmol) of Cy-g/L and a trace of Cy-dg were found. Cyanidin as aglycone of Cy-g and Cy-dg was not found in such plasma samples, neither were conjugated and methylated anthocyanins. The results indicated that anthocyanins are incorporated keeping structurally intact glycoside forms, from the digestive tract into the blood circulation system in mammals.     

Vegetative and reproductive dry weight inhibition in nitrogen‐ and phosphorus‐deficient Pima cotton 1

Whether the extent of dry weight inhibition by nitrogen (N) or phosphorus (P) deficiencies on different plant parts is the same and whether imposing moderate N and P deficiencies selectively suppress undesirable vegetative growth has not been studied in Pima cotton (Gossypium barbadense L.). The purpose of this study was to determine the extent to which dry matter accumulation in leaves, stems, and reproductive structures is inhibited by N and P deficiencies in Pima cotton. The study was conducted in 1991 and 1992 in a Uvalde silty clay loam soil (fine‐silty, mixed, hyperthermic Aridic Calciustolls). The treatments included applied rates of 0, 67, 135, 202, and 269 kg N ha^‐1 in a factorial combination with 0, 15, 29, and 44 kg P ha^‐1. Nitrogen deficiency (0 kg N ha^‐1) significantly (P≤0.05) reduced leaf (LDW) and stem (SDW) dry weights in both years and reproductive dry weight (RDW) in 1992. Nitrogen deficiency suppressed dry weight accumulation in leaves to a greater extent than in stems. Relative to 269 kg N ha^‐1, the 0 kg N ha^‐1 treatment resulted in a maximum LDW reduction of 62% at 144 DAP (days after planting) in 1991 and 36% at 121 DAP in 1992, compared with a corresponding SDW reduction of only 39% in 1991 and 25% in 1992. Dry weight accumulation in reproductive parts was the least affected by N deficiency. The decline in LDW associated with senescence and defoliation began earlier in treatments that received 0 or 67 kg N ha^‐1 than treatments that received ≥135 kg N ha^‐1. Phosphorus affected LDW and SDW in 1991, but its differential effect on LDW, SDW, and RDW was much smaller than that of N. Imposing a moderate level of N deficiency, not P deficiency, may be an effective Pima cotton management strategy to selectively suppress undesirable vegetative growth and enhance maturity.