Antioxidative activity of propolis extract in yeast cells

The antioxidative activities of propolis and its main phenolic compounds, caffeic acid, p-coumaric acid, ferulic acid, and caffeic acid phenethyl ester, were investigated in the yeast Saccharomyces cerevisiae. After 1 h of exposure of the yeast cells, their intracellular oxidation was measured using 2',7'-dichlorofluorescein. Yeast cells exposed to 96% ethanolic extracts of propolis in DMSO (EEP) showed decreased intracellular oxidation, with no significant differences seen for the individual phenolic compounds. However, cellular uptake was seen only for a moderately polar fraction of EEP (E2) and caffeic acid phenethyl ester. The EEP antioxidative activity thus resulted from this E2 fraction of EEP. The influence of EEP was also investigated at the mitochondrial proteome level, by analyzing its profile after 1 h of exposure of the yeast cells to EEP and E2. Changes in the levels of antioxidative proteins and proteins involved in ATP synthesis were seen.     

植物对铜的吸收运输及毒害机理研究进展

铜是植物生长发育必需的微量元素,但过量的铜对植物有毒害效应。因而植物体在长期的进化过程中形成了一套内稳态机制来调控铜离子在细胞中的浓度;既能满足细胞正常代谢对铜的需求又要使细胞免受铜毒害。植物对铜内稳态机理主要涉及植物对铜运输、螯合和区室化作用等方面,以此来达到对铜离子的吸收、分配和解毒的功能。本文综述了植物对铜的吸收运输与解毒的最新进展。     

Sodium copper chlorophyllin: in vitro digestive stability and accumulation by Caco-2 human intestinal cells

Sodium copper chlorophyllin (SCC), a mixture of water-soluble chlorophyll derivatives, is used as both a food colorant and a common dietary supplement. Although the potential antimutagenic and antioxidant properties of this commercial preparation have been demonstrated, limited information is available on its digestion and absorption by humans. Stability of SCC was examined during simulated gastric and small intestinal digestion. Three preparations were subjected to in vitro digestion: SCC in water, SCC in water + 10% corn oil, and SCC in applesauce. SCC components from raw material preparations and in digested samples were analyzed by C(18) HPLC with photodiode array detection. Cu(II)chlorin e(4), the major chlorin component of SCC, was relatively stable during simulated digestion. In contrast, greater than 90% of Cu(II)chlorin e(6) was degraded to undetermined products during digestion. Recovery of Cu(II)chlorin e(6) after digestion was increased by incorporation of SCC into applesauce, suggesting a protective role of the inclusion matrix for stabilization of labile SCC components. Accumulation of SCC derivatives was investigated by using differentiated cultures of the TC7 clone of the Caco-2 human intestinal cell line. Cellular accumulation from media containing 0.5 to 60 ppm SCC was linear with intracellular content ranging between 0.2 and 29.6 microg of total SCC per mg of cellular protein. Uptake of SCC by Caco-2 cells was significantly (p < 0.01) lower in cultures incubated at 4 degrees C than in those incubated at 37 degrees C. Although intracellular SCC was transported into both apical and basolateral compartments when Caco-2 cells were grown on inserts, apical efflux was significantly greater (p < 0.01) than basolateral efflux. Stability of Cu(II)chlorin e(4) during in vitro digestion and effective uptake by Caco-2 enterocyte-like cells support the likelihood that a portion of this SCC component or its metabolites is absorbed from the human intestine.     

Digestive Stability, micellarization, and uptake of beta-carotene isomers by Caco-2 human intestinal cells

While isomeric profiles of carotenoids found in food often differ from those in body fluids and tissues, insights about the basis for these differences remain limited. We investigated the digestive stability, relative efficiency of micellarization, and cellular accumulation of trans and cis isomers of beta-carotene (BC) using an in vitro digestion procedure coupled with human intestinal (Caco-2) cells. A meal containing applesauce, corn oil, and either water-soluble beadlets (WSB) or Dunaliella salina (DS) as a BC source was subjected to simulated gastric and small intestinal digestion. BC isomers were stable during digestion, and the efficiency of micellarization of cis-BC isomers exceeded that of all-trans-BC isomers. The cellular profile of carotenoids generally reflected that in micelles generated during digestion, and intracellular isomerization was minimal. These data suggest that cis isomers of BC are preferentially micellarized during digestion and transferred across the brush-border surface of the enterocyte from mixed micelles with similar efficiency as all-trans-BC at the concentrations of the carotenoids utilized in this study.     

GENOTYPIC DIFFERENCES IN PHOSPHORUS UPTAKE AND UTILIZATION OF WATERMELON UNDER LOW PHOSPHORUS STRESS

In this study, seven watermelon (Citrullus lanatus) cultivars were tested in solution culture experiment with limiting and ample phosphorus (P) supply to evaluate P uptake and utilization of watermelon under low P stress. Different genotypes showed considerable diversity in terms of biomass accumulation, P uptake, P utilization, root morphological parameters and photosynthetic parameters under low P stress. At low P supply, genotype XN8 and ZCHY were clearly superior to other genotypes in terms of total dry matter yield. The genotype ZJ has the highest P uptake ability, while the genotype XN8 has the highest P efficiency ratio and P utilization efficiency among the seven genotypes under low P stress. The P uptake ability of these genotypes was related significantly and positively to root morphological parameters and photosynthesis parameters under low P stress, the P utilization efficiency showed significant and positive correlation with total dry matter. Results showed existence of genetic differences among watermelon genotypes with regard to P absorption and utilization. The seven genotypes were classified into four groups: efficient responsive (ER), inefficient responsive (IER), efficient non-responsive (ENR) and inefficient nonresponsive (IENR) according to P utilization efficiency under low P stress and dry matter at high P supply. XN8 was identified as ER, and ZCHY was identified as ENR, which may be valuable resources for watermelon production in different soil with low P stress.     

Grape polyphenol resveratrol and the related molecule 4-hydroxystilbene induce growth inhibition, apoptosis, S-phase arrest, and upregulation of cyclins A, E, and B1 in human SK-Mel-28 melanoma cells

The effect of the naturally occurring polyphenol resveratrol (3,5,4'-trihydroxy-trans-stilbene; RES) on growth, cell cycle, and cyclins A, E, and B1 expression was investigated in the human SK-Mel-28 melanoma cell line. In addition, the structurally related compounds 4-hydroxy-trans-stilbene (4HST), piceatannol (3,5,3',4'-tetrahydroxy-trans-stilbene (PICE), and 4-trans-stilbenemethanol (4STMe) were also assayed in order to investigate the requirements of stilbenes to exert activity against melanoma cells. Both RES and 4HST inhibited cell growth in a dose- and time-dependent manner and upregulated the expression of cyclins A, E, and B1 with subsequent irreversible arrest of melanoma cells in the S-phase, concomitant with a decrease in G0/G1 and G2/M phases. In addition, potent apoptosis-mediated cell death was detected with the annexin V assay whereas no apoptosis was observed by flow cytometry, which encourages the assay of different methodologies to evaluate the effect of polyphenols on cell lines. The effect of PICE was not evaluated because of its instability in the reaction medium. No effect on cell cycle and cyclins expression was observed when 4STMe was assayed, which supported the critical requirement of the 4'-hydroxystyryl moiety to exert the above effects. In addition, this structural requirement also influenced the cellular uptake of stilbenes. The presence of two extra hydroxyl groups in RES increased its cytotoxicity whereas it diminished its efficiency to inhibit cell growth, upregulate cyclins expression, and arrest cell cycle in the S-phase with respect to 4HST. The present study suggests that the antimelanoma properties of dietary stilbenes, such as grape RES, cannot be ruled out, taking into account previous studies concerning the relationship between plasma and tissue concentrations and pharmacological activity of RES in animal models.     

Cytotoxicity of benzyl isothiocyanate in normal renal proximal tubular cells and its modulation by glutathione

In the present study, we examined the toxicity of benzyl ITC (BITC) and its urinary mercapturic acid metabolite (BITC-NAC), using a normal renal proximal tubular cell line, pig LLC-PK1. BITC increased cell death with an IC(50) value of about 7 μM, whereas the cytotoxic effect of BITC-NAC was five times weaker than that of BITC. We observed a significant necrosis of the compounds on LLC-PK1 cells with oxidative stress. In the presence of 5 mM glutathione (GSH), comparable to physiological levels, the cytotoxicity of BITC-NAC as well as BITC was significantly reduced. Furthermore, the increase in intracellular GSH levels by pretreatment with NAC before the BITC treatment resulted in inhibition of the BITC-induced necrotic events as well as intracellular oxidative stress. These results suggest that GSH is a determinant of cellular resistance against the BITC-mediated and oxidative stress-dependent cytotoxicity in renal proximal tubular cells.     

不同蔬菜对镉的吸收累积及亚细胞分布

采用营养液培养,基于叶绿体分离方法,研究了不同种类蔬菜对镉吸收、运输和亚细胞的分布规律。结果表明,蔬菜在不同浓度镉的营养液中培养1周后,蔬菜生物量没有产生显著差异,而不同种类的蔬菜生物量差异较大。镉在蔬菜叶片中大部分存在于细胞壁中,占总量的62％～85％,少量存在于原生质（不含叶绿体）和叶绿体中;随着营养液中镉浓度的增加,各组分中镉的含量明显增加,但分配比例变化不大。各种蔬菜根中镉的含量高于地上部镉含量;随着镉浓度的增加,根中镉分配比例从44％-59％降低至27％～38％;不同蔬菜根部对镉的吸收能力及镉向地上部转移的能力有显著性差异。     

Cellular uptake and efflux of trans-piceid and its aglycone trans-resveratrol on the apical membrane of human intestinal Caco-2 cells

Two stilbenes (trans-piceid and its aglycone trans-resveratrol) were investigated in the uptake across the apical membrane of the human intestinal cell line Caco-2 in order to determine their mechanisms of transport. The uptake was quantified using a reverse phase high-performance liquid chromatography method with fluorescence detection. The rate of cellular accumulation in the cells was found to be higher for trans-resveratrol than for trans-piceid. In addition, trans-resveratrol uses passive transport to cross the apical membrane of the cells, whereas the transport of trans-piceid is likely active. With regard to the mechanisms of transport, the involvement of the active transporter SGLT1 in the absorption of trans-piceid was deduced using various inhibitors directly or indirectly exploiting the activity of this transporter (glucose, phlorizin, and ouabain). Moreover, we investigated the involvement of the multidrug-related protein 2 (MRP2), an efflux pump present on the apical membrane, in stilbene efflux by Caco-2 cells. The effect of MK-571 (an MRP inhibitor) seems to implicate MRP2 as responsible for apical efflux of trans-piceid and trans-resveratrol.     

小牛血清对耐辐射奇球菌类胡萝卜素在肿瘤细胞中吸收的影响

耐辐射奇球菌类胡萝卜素deinoxanthin的抗氧化活性高于β-类胡萝卜素等,并且是参与耐辐射奇球菌极端抗性的重要物质。但是,deinoxanthin溶液在体外对光、氧等因素不稳定。本文旨在研究deinoxanthin的有效溶剂载体系统,并以此载体研究deinoxanthin对人卵巢癌细胞系A2780的药理学作用。通...     

蚯蚓中菲分布的分层次方法研究

The distribution of heavy metals in earthworms has been widely studied, highlighting the importance of the fate of these metals.However, little information is available on the distribution of hydrophobic organic contaminants(HOCs) within earthworms. The aim of this study was to propose a hierarchic method to study the distribution of phenanthrene(PHE), a typical HOC, in Eisenia fetida at several levels: sub-organism(pre-clitellum, clitellum and post-clitellum), tissue(body wall, gut and body fluid) and subcellular(intracellular and extracellular fractions). Earthworms were incubated in the soils amended with low(LC, 10 mg kg-1) and high concentrations(HC, 50 mg kg-1) of PHE and sampled at different time intervals. At the sub-organism level, the distribution of PHE was homogeneous among the sub-organism fractions in the LC treatment but heterogeneous in the HC treatment and gradually reached the following form of post-clitellum ≈ clitellum pre-clitellum. The uptake and elimination kinetics of PHE in the sub-organism were well described by a one-compartment model. At the tissue level, the concentration of PHE followed the order of gut body fluid body wall; while at the subcellular level, the concentration of PHE in the extracellular fraction was 1.23 to 4.68 times higher than that in the intracellular fraction. Therefore, the simple circulatory system of earthworms may account for the PHE distribution at the sub-organism level. Partition pathways(passive diffusion) of PHE between the body wall, body fluid and gut as well as the processes of PHE entrance into the inner cellular compartment via passive diffusion were experimentally supported.     

Removal of carbaryl, linuron, and permethrin by Lupinus angustifolius under hydroponic conditions

The metabolism of organic pollutants by plants normally requires contaminant direct uptake by cells. Factors affecting this uptake and the later distribution of chemicals within the plant include the physicochemical properties of the compounds (concentration, structure, solubility, log k(ow), diffusion rate) and the biochemical characteristics of the plant. This paper reports the tolerance, uptake, and effects of the pesticides carbaryl, linuron, and permethrin on Lupinus angustifolius germination and growth as well as contaminant intraplant distribution and possible degradation. Lupine plants were grown in hydroponic culture containing either 1 or 5 mg of the individual pesticides, or combinations of these (1, 5, or 10 mg of each), in 100 mL nutrient and water solutions. Analysis of the remaining solutions 8 days post-germination showed the water solutions to have higher remaining pesticide concentrations than nutrient solutions. Furthermore, in the presence of pesticides, germination was more frequent in the water solutions. After 16 days of growth, the plants were harvested, and their tissues were microwaved digested and analyzed by reversed-phase liquid chromatography. Although only minor quantities of each pesticide were detected in plant tissues, their amount in the roots was higher than in the stems. No accumulation was noted in the cotyledons, and only 2% of linuron was detected in the leaves. Mass recovery at the end of the experiment showed that 57, 53, and 55% of carbaryl, linuron, and permethrin, respectively, were degraded and/or bound in an irreversible manner to plant material. The results suggest that L. angustifolius could be useful for the cleaning/remediation of pesticide-contaminated water.     

Neuroprotective effects of the citrus flavanones against H2O2-induced cytotoxicity in PC12 cells

The citrus flavanones hesperidin, hesperetin, and neohesperidin are known to exhibit antioxidant activities and could traverse the blood-brain barrier. H2O2 formation induces cellular oxidative stress associated with neurodegenerative diseases. In this study, protective effects of pretreatments (6 h) with hesperidin, hesperetin, and neohesperidin (0.8, 4, 20, and 50 microM) on H2O2-induced (400 microM, 16 h) neurotoxicity in PC12 cells were evaluated. The results showed that hesperetin, hesperidin, and neohesperidin, at all test concentrations, significantly ( p < 0.05) inhibited the decrease of cell viability (MTT reduction), prevented membrane damage (LDH release), scavenged ROS formation, increased catalase activity, and attenuated the elevation of intracellular free Ca2+, the decrease of mitochondrial membrane potential (except those of 0.8 microM neohesperidin-treated cells) and the increase of caspase-3 activity in H2O2-induced PC12 cells. Meanwhile, hesperidin and hesperetin attenuated decreases of glutathione peroxidase and glutathione reductase activities and decreased DNA damage in H2O2-induced PC12 cells. These results first demonstrate that the citrus flavanones hesperidin, hesperetin, and neohesperidin, even at physiological concentrations, have neuroprotective effects against H2O2-induced cytotoxicity in PC12 cells. These dietary antioxidants are potential candidates for use in the intervention for neurodegenerative diseases.     

锌与氮磷钾配合喷施对小麦锌累积、分配及转移的影响

【目的】 在作物上喷施锌肥与目前应用较广泛的喷施大量元素肥料 (氮、磷或钾肥) 相配合,是解决人体锌缺乏问题的重要途径。本研究初步阐明了锌与氮、磷及钾肥配合喷施对小麦籽粒锌营养品质的影响机制,为小麦籽粒有效可行富锌方法提供理论参考。 【方法】 2010—2014年连续进行了4年裂区田间试验。主处理为不同氮肥用量土施 (N 0、120和240 kg/hm2),副处理为喷蒸馏水 (CK)、喷0.3% ZnSO₄ (Zn)、喷0.3% ZnSO₄和1.7%尿素 (Zn + N)、喷0.3% ZnSO₄和0.2% KH₂PO₄ (Zn + P + K)、喷0.3% ZnSO₄和0.5% K₂SO₄ (Zn + K)。分析测定小麦开花期地上部以及成熟期各部位 (籽粒、叶片、颖壳和茎秆) 锌含量,研究锌与氮、磷或钾肥配合喷施对锌在成熟期小麦各部位的分配,以及花前和花后锌分配比例及其对籽粒锌累积的表观贡献率的影响。 【结果】 喷施Zn、Zn + N、Zn + P + K或Zn + K处理后锌含量提升幅度以叶片最大 (2.4～7.7倍),颖壳 (2.0～4.7倍) 和籽粒 (1.8～2.4倍) 次之,茎秆最小 (0.2～1.0倍),锌在叶片和颖壳分配比明显提高。与单喷Zn相比,Zn + N或Zn + K处理籽粒和叶片锌含量进一步增加,而Zn + P + K处理籽粒和叶片锌含量均有所降低。与Zn处理相比,花后营养器官锌吸收量及其向籽粒的转移量在喷Zn + N时分别增加12和14 g/hm2,在喷Zn + K时增加44和32 g/hm2,但喷Zn + P + K时分别降低37和18 g/hm2。土施氮肥亦可显著增加籽粒和各营养器官锌含量,以及锌在营养器官的累积和再转移,但增幅明显低于各喷锌处理。此外,与Zn处理相比,Zn + N、Zn + P + K或Zn + K处理未进一步影响锌在小麦各部位的分配,但锌肥的回收率在Zn + N或Zn + K处理下显著提高,在Zn + P + K处理下显著降低。 【结论】 锌肥与氮肥或钾肥配合喷施,主要通过增加营养器官对锌的吸收及向籽粒的转移量,进一步提高籽粒锌含量,而磷锌配合喷施通过降低营养器官对锌的吸收及向籽粒的转移进而降低籽粒锌含量。      

Antioxidant and prooxidant actions of prenylated and nonprenylated chalcones and flavanones in vitro

Prenylated flavonoids found in hops and beer, i.e., prenylchalcones and prenylflavanones, were examined for their ability to inhibit in vitro oxidation of human low-density lipoprotein (LDL). The oxidation of LDL was assessed by the formation of conjugated dienes and thiobarbituric acid-reactive substances (TBARS) and the loss of tryptophan fluorescence. At concentrations of 5 and 25 microM, all of the prenylchalcones tested inhibited the oxidation of LDL (50 microg protein/ml) induced by 2 microM copper sulfate. The prenylflavanones showed less antioxidant activity than the prenylchalcones, both at 5 and 25 microM. At 25 microM, the nonprenylated chalcone, chalconaringenin (CN), and the nonprenylated flavanone, naringenin (NG), exerted prooxidant effects on LDL oxidation, based on TBARS formation. Xanthohumol (XN), the major prenylchalcone in hops and beer, showed high antioxidant activity in inhibiting LDL oxidation, higher than alpha-tocopherol and the isoflavone genistein but lower than the flavonol quercetin. When combined, XN and alpha-tocopherol completely inhibited copper-mediated LDL oxidation. These findings suggest that prenylchalcones and prenylflavanones found in hops and beer protect human LDL from oxidation and that prenylation antagonizes the prooxidant effects of the chalcone, CN, and the flavanone, NG.     

Effects of Water Acidity and Metal Concentration on Accumulation and Within-Plant Distribution of Metals in the Aquatic Bryophyte Fontinalis antipyretica

With the aim of investigating the effects of wateracidity and metal concentrations on the accumulationof heavy metals by aquatic bryophytes, batches of Fontinalis antipyretica Hedw. were transplanted froma clean site to one of four sites with characteristicsranging from clean to severely contaminated (highacidity and high heavy metal concentrations). After 1–35 days, metal levels were determined in theextracellular and intracellular compartments of thetransplanted material. The results show that heavymetal uptake is clearly lower in acid waters than innear-neutral waters, possibly due to competitivedisplacement of metal cations from extracellularbinding sites and membrane transport proteins byprotons. The reduced accumulation is particularlymarked in the extracellular compartment; theintracellular compartment shows rapid accumulation oflarge amounts of metal, followed by equally rapidrelease. The effects of heavy metals and acidity onphysiology were also investigated, on the basis oflevels of the essential cations potassium (K),magnesium (Mg) and calcium (Ca) in the extracellularand intracellular compartments. Bryophytestransplanted to the severely contaminated sites showedloss of intracellular K (indicating altered membranepermeability), loss of extracellular Mg, and loss ofboth extra- and intracellular Ca (implying damage tothe cell wall).     

Curcuminoids-cellular uptake by human primary colon cancer cells as quantitated by a sensitive HPLC assay and its relation with the inhibition of proliferation and apoptosis

Curcumin, which is a bright orange-yellow pigment of turmeric with antioxidant properties, has been shown to produce a potent preventative action against several types of cancers in recent studies. It has also been reported to protect the development of colon tumor in animals being fed with carcinogen. In the colon cancer cells, curcumin was illustrated to inhibit cell proliferation and induce apoptosis. As an antioxidant, it acts as an anti-inflammatory as well as an antitumor agent. Curcumin has been detected to exist in nature in the form of curcuminoids, a mixture of curcumin, the major component, with two of its related demethoxy compounds (demethoxycurcumin and bisdemethoxycurcumin). In the present study, we have investigated the antiproliferation and induced apoptosis effects of curcuminoids on colon cancer, using the primary cancer cells isolated from Taiwanese colon cancer patients as the model for colorectal cancer. Results showed that curcuminoids inhibited cell proliferation and induced apoptosis of these human primary colon cancer cells. The effects were observed in a dose-dependent manner as dose increased from 12.5 to 100 microM. With the aim of furthering the fundamental understanding of the mechanisms underlying the antiproliferation and induced apoptosis effects of curcuminoids on these human colon cancer cells, we developed a sensitive, rapid, and reproducible assay method based on high-performance liquid chromatography (HPLC). This HPLC technique developed was found to successfully determine, in a quantitative manner, the cellular uptake of curcuminoids. The uptake of these curcuminoids by the colon cancer cells was shown to increase as the dose of curcuminoids was increased. The observations of inhibited proliferation and increased apoptosis in the colon cancer cells appeared to be associated with the cellular uptake of curcuminoids.     

Inulin affects iron dialyzability from FeSO4 and FeEDTA solutions but does not alter Fe uptake by Caco-2 cells

The in vitro effects of inulin on the fluxes of Fe (F(Fe)) and uptake by Caco-2 cells from FeSO4 and FeEDTA were evaluated. Cell ferritin formation was used as a measure of Fe uptake. Mitochondrial (MTT test) and lysosomal activities were monitored as biomarkers of the changes of cellular metabolism. Changes in mRNA expression of Fe transporters, DMT1 and Dcytb, were evaluated. Inulin decreased dialyzability and F(Fe) from FeSO4 solution, suggesting a mineral binding effect, but increased those from FeEDTA. Cultures exposed to FeEDTA solutions exhibited higher ferritin values and MTT conversion percentages. Regardless of Fe source, cell Fe uptake and mRNA expression of Fe transporters were similar with or without inulin, suggesting that inulin did not impair Fe uptake. These observations might indicate a faster cellular Fe internalization from FeEDTA solutions. From a physiological perspective, the decreased F(Fe) from FeSO4 might be reflected in a decreased Fe uptake.     

厌氧去除活性污泥中痕量有机污染物:15年实验研究

Knowledge of cellular metal homeostasis will provide a better understanding of the mechanisms involved in metal tolerance and hyperaccumulation in metal-hyperaccumulating plants. Energy dispersive X-ray spectrometry (EDS) was used to determine the localization of cadmium (Cd) in leaves of the Zn/Cd hyperaccumulator Picris divaricata which had a shoot Cd concentration of 565 mg kg 1 after 2 weeks of growth in solution culture supplying 10 μmol L-1 CdCl2 . The results indicated that Cd was distributed mainly in the trichomes, upper and lower epidermis and bundle sheath cells, with a relatively low level of Cd in mesophyll cells. Mesophyll protoplasts isolated from leaves remained viable after 24 h exposure to CdCl2 at a concentration up to 1 mmol L-1 , indicating their high tolerance to Cd. The intracellular Cd was visualized by staining with Leadmium Green dye, a cellular permeable Cd fluorescence probe. The results showed that the majority of protoplasts (> 82%) did not accumulate Cd, with only a minority (< 18%) showing Cd accumulation. In the Cd-accumulating protoplasts, Cd accumulation was depressed by the addition of Fe2+ , Mn2+ and the metabolic inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP), but not by Ca 2+ or Zn2+ . Furthermore, the entire process of Cd uptake from external solution into the cytoplasm and subsequent sequestration into vacuoles was successfully recorded by confocal images. These results suggested that reduced cellular Cd accumulation and efficient Cd vacuolar sequestration in mesophyll cells might be responsible for cellular Cd tolerance and distribution in the leaves of P. divaricata.     

Effect of hesperetin against oxidative stress via ER- and TrkA-mediated actions in PC12 cells

Hesperetin is known to activate estrogen receptors (ERs). Estrogen-mediated neuroprotection could be via both ER and tyrosine kinase receptor (Trk) signaling. This study tested whether hesperetin protected PC12 cells from hydrogen peroxide induced oxidative damage via ER- and/or TrkA-mediated actions. Hesperetin (0.1, 1, and 50 μM) inhibited cell viability decreases and reactive oxygen species, intracellular calcium level, and caspase-3 activity increases in H(2)O(2)-induced PC12 cells. Such actions were significantly (p < 0.05) suppressed by ICI 182,780 (an ER antagonist) or K252a (a TrkA antagonist) at low concentrations (0.1 or 1 μM) only. Hesperetin also stimulated the activation of Akt, ERK, and CREB as well as induced brain-derived neurotrophic factor, PPARγ coactivator 1α (PGC-1α), and seladin-1 (selective Alzheimer's disease indicator-1) via both ER and TrkA in the cells. This study demonstrates that the neuroprotective effects of hesperetin, at low concentrations, are attributed to its stimulation on receptor signaling. Moreover, ER and TrkA are known to be expressed in most Alzheimer's disease (AD) vulnerable brain regions. This study thus suggests that hesperetin might have potential for intervention in neurodegenerative disorders, particularly for AD.