Benzyl isothiocyanate (BITC) induces G2/M phase arrest and apoptosis in human melanoma A375.S2 cells through reactive oxygen species (ROS) and both mitochondria-dependent and death receptor-mediated multiple signaling pathways

Benzyl isothiocyanates (BITC), a member of the isothiocyanate (ITC) family, inhibits cell growth and induces apoptosis in many types of human cancer cell lines. The present study investigated mechanisms underlying BITC-induced apoptosis in A375.S2 human melanoma cancer cells. To observe cell morphological changes and viability, flow cytometric assays, cell counting, and a contrast-phase microscopic examination were carried out in A375.S2 cells after BITC treatment. Cell cycle distribution and apoptosis were assessed with the analysis of cell cycle by flow cytometric assays, DAPI staining, propidium iodide (PI), and annexin V staining. Apoptosis-associated factors such as reactive oxygen species (ROS) formation, loss of mitochondrial membrane potential (ΔΨ(m)), intracellular Ca(2+) release, and caspase-3 activity were evaluated by flow cytometric assays. Abundance of cell cycle and apoptosis associated proteins was determined by Western blotting. AIF and Endo G expression was examined by confocal laser microscope. Results indicated that (1) BITC significantly reduced cell number and induced cell morphological changes in a dose-dependent manner in A375.S2 cells; (2) BITC induced arrest in cell cycle progression at G(2)/M phase through cyclin A, CDK1, CDC25C/Wee1-mediated pathways; (3) BITC induced apoptosis and increased sub-G(1) population; and (4) BITC promoted the production of ROS and Ca(2+) and loss of ΔΨ(m) and caspase-3 activity. Furthermore, BITC induced the down-regulation of Bcl-2 expression and induced up-regulation of Bax in A375.S2 cells. Moreover, BITC-induced cell death was decreased after pretreatment with N-acetyl-l-cysteine (NAC, a ROS scavenger) in A375.S2 cells. In conclusion, the results showed that BITC promoted the induction of G(2)/M phase arrest and apoptosis in A375.S2 human melanoma cells through ER stress- and mitochondria-dependent and death receptor-mediated multiple signaling pathways. These data suggest that BITC has potential as an agent for the treatment of melanoma.     

Oligophosphopeptides derived from egg yolk phosvitin up-regulate gamma-glutamylcysteine synthetase and antioxidant enzymes against oxidative stress in Caco-2 cells

Previously, we have found phosphopeptides (PPPs) from hen egg yolk phosvitin possess a potent antioxidative activity against oxidative stress in human intestinal epithelial cells, Caco-2. However, their biological activity at the cellular level has not yet fully understood. The objective of this study is to evaluate the regulation of glutathione (GSH) biosynthesis-associated and antioxidant enzymes against oxidative stress in Caco-2 cells using an in vitro model. Treatment of 1 mM H2O2-induced Caco-2 cells with PPPs increased cellular GSH levels, concomitant with a significant increase in gamma-glutamylcysteine synthetase (gamma-GCS) activity and the expression of gamma-GCS heavy subunit mRNA. Furthermore, intracellular glutathione reductase, glutathione S-transferase, and catalase activities were elevated by PPPs. In addition, PPPs with high content of phosphorus showed higher induction of these enzyme activities than PPPs without phosphorus. These data indicate that oligophosphopeptides from hen egg yolk phosvitin can up-regulate cellular GSH biosynthesis-associated enzymes activity and antioxidative activities, which play key roles against tissue oxidative stress in the human intestinal epithelial cells.     

Antioxidative activity of amino acids on tissue oxidative stress in human intestinal epithelial cell model

The protective effects of amino acids against H 2O 2-induced oxidative stress were investigated in an in vitro assay using human intestinal epithelial cells. Caco-2 cells were pretreated with amino acids (1, 2, and 5 mM) for 2 h and then stimulated with 1 mM H 2O 2 for 6 h. The secretion of IL-8, a proinflammatory mediator, was determined by ELISA as an indicator of tissue oxidative stress. The inhibition of H 2O 2-induced IL-8 secretion from Caco-2 cells was observed by pretreatment with Cys, Val, Ile, Leu, Trp, His, Lys, and Ala. Cys enhanced glutathione (GSH) biosynthesis enzyme activity and increased cellular GSH levels. Branched-chain amino acids such as Val, Ile, and Leu elevated activities of GSH S-transferase (GST) and catalase. Trp, His, and Lys caused increases in GST activity. Ala enhanced GSH reductase activity. These data suggest that specific amino acids exert protective effects against tissue oxidative stress in intestinal epithelial cells based on the structure.     

Effect of ethanol and red wine on ochratoxin a-induced experimental acute nephrotoxicity

Ochratoxin A (OTA), is a nephrotoxic mycotoxin present in wine, which is nephrotoxic in humans. Our working hypothesis is that natural substances in wine may counteract OTA toxicity. Thirty-six rats were randomized to OTA dissolved in saline, red wine, or 13.5% ethanol or to OTA-free wine, ethanol, or saline. OTA (289 microg/kg of body weight/48 h) was administered by gastric gavage for 2 weeks. Serum creatinine, tubular enzymuria, renal lipohydroperoxides (LOOH), reduced (GSH) and oxidized (GSSG) glutathione, and renal superoxide dismutase activity (SOD) were determined in renal tissue. OTA alone produced significant increases in renal lipoperoxides and significant decreases in SOD and GSH/GSSG ratio. In red wine or ethanol, OTA was less nephrotoxic, reducing oxidative damage as revealed by LOOH. In OTA-wine and OTA-ethanol groups, SOD activity was higher than in the OTA-treated one, suggesting that both ethanol and nonalcoholic fractions may preserve antioxidant reserve. GSH/GSSG ratio was significantly preserved only in the OTA-wine group and not in OTA-ethanol. Red wine may exert a protective effect against OTA nephrotoxicity by limiting oxidative damage. The ostensible protection afforded by ethanol deserves further investigation.     

Alkyl hydroxytyrosyl ethers show protective effects against oxidative stress in HepG2 cells

Alkyl hydroxytyrosyl ethers (methyl, ethyl, propyl, and butyl ethers) have been synthesized from hydroxytyrosol (HTy) in response to the increasing food industry demand of new lipophilic antioxidants. Having confirmed that these compounds reach portal blood partially unconjugated and thus are effectively absorbed, their potential antioxidant activity was evaluated in the human hepatocarcinoma cell line (HepG2). The effects of 0.5-10 μM alkyl hydroxytyrosyl ethers on HepG2 cell integrity and redox status were assessed as well as the protective effect against oxidative stress induced by tert-butylhydroperoxide (t-BOOH). Cell viability (Crystal violet) and cell proliferation (BrdU assay) were measured as markers of cell integrity, concentration of reduced glutathione (GSH), generation of reactive oxygen species (ROS), and activity of antioxidant enzymes glutathione peroxidase (GPx) and glutathione reductase (GR) as markers of redox status and determination of malondialdehyde (MDA) as a marker of lipid peroxidation. Direct treatment of HepG2 with alkyl hydroxytyrosyl ethers induced slight changes in cellular intrinsic antioxidants status, reducing ROS generation and inducing changes in GPx and GR activities. Pretreatment of HepG2 cells with alkyl hydroxytyrosyl ethers counteracted cell damage induced by t-BOOH, partially after 2 h and completely after 20 h, by increasing GSH and decreasing ROS generation, MDA levels, and antioxidant enzyme (GPx and GR) activity. According to these results the alkyl hydroxytyrosyl ethers show clear protective effects against oxidative stress, related to their lipophilic nature, that are similar to or even higher than those of their precursor, HTy.     

Effects of polyphenolic compounds on tumor necrosis factor-α (TNF-α)-induced changes of adipokines and oxidative stress in 3T3-L1 adipocytes

Over the last few decades, obesity has become a global epidemic in both developed and developing countries. Recent studies have indicated that obesity is closely associated with chronic inflammation characterized by abnormal levels of adipocytokines and inflammatory cytokines in adipocytes. The aim of this work was to study the effects of 21 polyphenolic compounds on tumor necrosis factor-α (TNF-α)-induced changes of adipokines and oxidative stress in 3T3-L1 adipocytes. The results showed that p-coumaric acid, quercetin, and resveratrol have greater inhibition (p < 0.05) of a TNF-α-induced increase in the production of interleukin-6 (IL-6) among 21 tested polyphenolic compounds. p-Coumaric acid, quercetin, and resveratrol demonstrated inhibitions of TNF-α-induced changes in levels of monocyte chemoattractant protein-1 (MCP-1), plasminogen activator inhibitor-1 (PAI-1), and intracellular reactive oxygen species (ROS) in 3T3-L1 adipocytes. Furthermore, p-coumaric acid, quercetin, and resveratrol increased levels (p < 0.05) of secreted adiponectin, superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPx), and glutathione S-transferase (GST) in TNF-α-treated 3T3-L1 adipocytes. These results indicate that the inhibition of TNF-α-induced changes of adipokines and oxidative stress by some polyphenolic compounds might have further implications in preventing obesity-related pathologies.     

Benzylglucosinolate, benzylisothiocyanate, and myrosinase activity in papaya fruit during development and ripening

Papaya is a climacteric fruit that has high amounts of benzylglucosinolates (BG) and benzylisothiocyanates (BITC), but information regarding levels of BG or BITC during fruit development and ripening is limited. Because BG and BITC are compounds of importance from both a nutritional and a crop yield standpoint, the aim of this work was to access data on the distribution and changes of BG and BITC levels during fruit development and ripening. BG and BITC levels were quantified in peel, pulp, and seeds of papaya fruit. Volatile BITC was also verified in the internal cavity of the fruit during ripening. The influence of the ethylene in BG and BITC levels and mirosinase activity was tested by exposing mature green fruits to ethylene and 1-methylcyclopropene (1-MCP). The highest BG levels were detected in seeds, followed by the peel and pulp being decreased in all tissues during fruit development. Similarly, the levels of BITC were much higher in the seeds than the peel and pulp. The levels of BG for control and ethylene-treated fruit were very similar, increasing in the pulp and peel during late ripening but not changing significantly in seeds. On the other hand, fruit exposed to 1-MCP showed a decrease in BG amount in the pulp and accumulation in seed. The treatments did not result in clear differences regarding the amount of BITC in the pulp and peel of the fruit. According to the results, ethylene does not have a clear effect on BITC accumulation in ripening papaya fruit. The fact that BG levels in the pulp did not decrease during ripening, regardless of the treatment employed, and that papaya is consumed mainly as fresh fruit, speaks in favor of this fruit as a good dietary source for glucosinolate and isothiocyanates.     

Nanoemulsions of cancer chemopreventive agent benzyl isothiocyanate display enhanced solubility, dissolution, and permeability

Benzyl isothiocyanate (BITC), a compound found in cruciferous vegetables, is an effective chemopreventive agent. The objective of this study was to develop nanoemulsion formulations for the oral delivery of BITC. Optimized oil-in-water BITC nanoemulsions were prepared by a spontaneous self-nanoemulsification method and a homogenization-sonication method. Both nanoemulsions entrapped high amounts of BITC (15-17 mg/mL), with low polydispersity and good colloidal stability. The BITC nanoemulsions showed enhanced solubility and dissolution compared to pure BITC. These formulations markedly increased the apical to basolateral transport of BITC in Caco-2 cell monolayers. The apparent permeability values were 3.6 × 10(-6) cm/s for pure BITC and (1.1-1.3) × 10(-5) cm/s for BITC nanoemulsions. The nanoemulsions were easily taken up by human cancer cells A549 and SKOV-3 and inhibited tumor growth in vitro. This work shows for the first time that BITC can be formulated into nanoemulsions and may show promise in enhancing absorption and bioavailability.     

Cellular Impact of Metal Trace Elements in Terricolous Lichen Diploschistes muscorum (Scop.) R. Sant. – Identification of Oxidative Stress Biomarkers

The purpose of this research is to present toxic effects of some heavy metals (cadmium, lead and zinc) on lichen physiology. In the North of France, those metals are concentrated for example in industrial polluted soils near metallurgic plants and waste dumps. Our investigations were conducted on Diploschistes muscorum (Scop.) R. Sant, a terricolous lichen growing on previously quoted soils as well as on non-contaminated ones. Different stress parameters were investigated as potassium leakage – which is related to the loss of membrane integrity – and oxidative stress through following parameters: malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD). It seems, therefore, that heavy metals induce oxidative stress in this lichen, in which we found membrane damage and enhancement of SOD activity and GSH concentrations. With regard to very high concentrations of Cd, Pb and Zn, a resistance involving antioxidant mechanisms limits the expected damage. Those mechanisms involve GSH, SOD as well as secondary metabolites, which have this resistance property. This study is a first step of investigation to use – in the future– physiological parameters as tool for environment assessment.     

6-shogaol (alkanone from ginger) induces apoptotic cell death of human hepatoma p53 mutant Mahlavu subline via an oxidative stress-mediated caspase-dependent mechanism

Mahlavu cells, poorly differentiated and p53 mutants of a human hepatoma subline, are known to be highly refractory to a number of chemotherapeutic agents and radiotherapy due to their high expressions of multidrug resistance gene-1 (MDR-1) and Bcl-2 proteins. Thus, it is desirable to search for an alternative strategy for effective eradication of this type of cancer cells. We present evidence here for the first time that 6-shogaol (6-SG), an alkanone isolated from the rhizomes of ginger, can effectively induce apoptotic cell death of Mahlavu cells via an oxidative stress-mediated caspase-dependent mechanism. The cascade of events in 6-SG-induced apoptosis of these cells involved an initial overproduction of reactive oxygen species (ROS) followed by a severe depletion of intracellular glutathione (GSH) contents. Both events consequently entailed a significant drop in mitochondrial transmembrane potential (DeltaPsim), which ultimately activated the activities of caspases 3/7 resulting in the DNA fragmentation. Interestingly, we also found that N-acetylcysteine (NAC), an antioxidant and a precursor of GSH biosynthesis, could offer a near complete protection of apoptotic cell death exerted by 6-SG. Similarly, exogenously added GSH could also provide protection with an equal efficacy. However, it was paradoxical that both Boc-Asp(OMe)-fmk (a broad caspases inhibitor) and cyclosporin A (an mitochondrial permeability transition opening inhibitor) could only partially protect these cells from 6-SG-induced apoptosis. Taking these data into consideration, it is obvious that GSH depletion is the major contributing factor in arbitrating 6-SG-induced apoptosis of Mahlavu cells. In conclusion, we provide here a novel modality that can help to eradicate a p53 mutant of human hepatoma cells by using a natural consistent isolated form of ginger. These data also provide evidence to reaffirm the notion that consumption of certain foodstuffs can be beneficial to health because some of the constituents contained in them may be anticarcinogenic.     

Synthesis and biological evaluation of novel C(6) modified baicalein derivatives as antioxidative agents

Baicalein, one of the major flavones, was found to be responsible for the antioxidative activity of the traditional Chinese medicinal herb Huang-Qin ( Scutellaria baicalensis Georgi), which is widely used as an antioxidative, anti-inflammatory, and antitumor agent. The hydroxyl group of the A ring of the baicalein was alkylated at position 6 with terpenoids such as prenyl, geranyl, and farnesyl groups, and their free radical scavenging activities and glutathione (GSH) depletion capacities were examined. Their free radical scavenging activity was measured according to the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(*+)) scavenging method. Baicalein and newly synthesized baicalein derivatives were found to be good free radical scavengers. Flow cytometrical method was employed to measure the intracellular antioxidative activity and GSH depletion capacity of these derivatives in human acute monocytic leukemia cell line (THP-1). It was also found that baicalein and its derivatives could decrease the levels of exogenous cumene hydroperoxide and H2O2 in THP-1 cells. These compounds also could significantly inhibit the intracellular GSH depletion induced by cumene hydroperoxide in THP-1 cells. The production of cumene hydroperoxide-induced Bax, a pro-apoptotic related protein, could also be inhibited by baicalein and its derivatives. These results suggested that baicalein and its derivatives could be beneficial to human health.     

Effect of curcumin on inflammation and oxidative stress in cisplatin-induced experimental nephrotoxicity

Nephrotoxicity is a major complication and a dose limiting factor for cisplatin therapy. Recent evidence suggests that inflammation and oxidative stress may contribute to the pathogenesis of cisplatin-induced acute renal failure. Curcumin is claimed to be a potent anti-inflammatory and antioxidant agent. The present study was performed to explore the effect of curcumin against cisplatin-induced experimental nephrotoxicity. Curcumin in the dosages of 15, 30, and 60 mg kg(-1) was administered 2 days before and 3 days after cisplatin administration. Renal injury was assessed by measuring serum creatinine, blood urea nitrogen, creatinine, urea clearance, and serum nitrite levels. Renal oxidative stress was assessed by determining renal malondialdehyde levels, reduced glutathione levels and enzymatic activities of superoxide dismutase and catalase. Systemic inflammation was assessed by tumor necrosis factor-alpha (TNF-alpha) levels. A single dose of cisplatin resulted in marked inflammation (486% rise in TNF-alpha level) and oxidative stress and significantly deranged renal functions as well as renal morphology. The serum TNF-alpha level was markedly reduced in curcumin-treated rats. Curcumin treatment significantly and dose-dependently restored renal function, reduced lipid peroxidation, and enhanced the levels of reduced glutathione and activities of superoxide dismutase and catalase. The present study demonstrates that curcumin has a protective effect on cisplatin-induced experimental nephrotoxicity, and this effect is attributed to its direct anti-inflammatory and strong antioxidant profile. Hence, curcumin has a strong potential to be used as a therapeutic adjuvant in cisplatin nephrotoxicity.     

Antioxidative stress activity of oligophosphopeptides derived from hen egg yolk phosvitin in Caco-2 cells

The protective effects of hen egg yolk phosvitin phosphopeptides (PPPs) against hydrogen peroxide (H2O2)-induced oxidative stress were evaluated in an in vitro assay using human intestinal epithelial cells. Caco-2 cells were stimulated with 1 mM H2O2 for 6 h, and the secretion of IL-8, a proinflammatory mediator, was determined by ELISA as a biomarker of oxidative stress. The inhibition of H2O2-induced IL-8 secretion from Caco-2 cells was observed by pretreatment for 2 h with PPPs, but not with phosvitin. PPPs also suppressed the formation of malondialdehyde in H2O2-treated Caco-2 cells. Furthermore, intracellular glutathione levels and glutathione reductase activity were elevated by the addition of PPPs. The protective effects of PPPs against H2O2-induced oxidative stress were almost the same as that of glutathione, and PPPs with a high content of phosphorus exhibited higher protective activity than PPPs without phosphorus; however, phosphoserine itself did not show any significant antioxidative stress activity. These findings suggest that oligophosphopeptides from hen egg yolk phosvitin possess novel antioxidative activity against oxidative stress in intestinal epithelial cells and that phosphorus and peptide structure seem to have a key role in the activity.     

Cadmium‐induced oxidative stress and protection by L‐Galactono‐1, 4‐lactone in winter wheat (Triticum aestivum L.)

Two hydroponic culture experiments were conducted to investigate cadmium (Cd)‐induced oxidative stress in winter wheat (Triticum aestivum L.) seedlings and the effects of L‐Galactono‐1, 4‐lactone (GalL), the biosynthetic precursor of the antioxidant ascorbate (AsA), on the oxidative stress induced by Cd. In experiment 1, with application of Cd (0, 10, 25, 50 µM) in nutrient solution, hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) levels as well as membrane permeability in both shoots and roots were significantly increased, indicating Cd‐induced oxidative stress and lipid peroxidation as well as plasma‐membrane damage in the plants. In experiment 2, H₂O₂ levels in plants exposed to Cd were significantly reduced by the addition of GalL (25 mM), associated with increased activities of peroxidase (POD), indicating that GalL alleviated the oxidative stress induced by Cd. Unexpectedly, however, the MDA levels were not reduced by the addition of GalL. Does Cd also induce lipid peroxidation directly besides via formation of reactive oxygen species (ROS)? This needs further study.     

Oxidative Status of Stressed Caenorhabditis elegans Treated with Epicatechin

The aim of this work was to examine the mechanisms involved in the in vivo antioxidant effects of epicatechin (EC), a major flavonoid in the human diet. The influence of EC in different oxidative biomarkers (reactive oxygen species (ROS) production, intracellular glutathione, activity of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD)) was studied in the model organism Caenorhabditis elegans . Under thermal stress condition, exposure of the worms (wild type N2 strains) to EC (200 μM) significantly reduced ROS levels (up to 28%) and enhanced the production of reduced glutathione (GSH). However, no significant changes were appreciated in the activities of GPx, CAT, and SOD, suggesting that further activation of these antioxidant enzymes was not required once the concentration of ROS in the EC-treated worms was restored to what could be considered physiological levels.     

Identification of cytotoxic constituents of Narthecium ossifragum using bioassay-guided fractionation

Narthecium ossifragum, a member of the Liliaceae family, contains phytochemicals that have hepatotoxic and nephrotoxic activity in several ruminant species. 3-Methoxyfuran-2(5H)-one has previously been isolated as the principal nephrotoxin, and its toxicity has been confirmed in vivo. However, previous investigations into the nephrotoxicity of N. ossifragum both in vivo and in vitro indicate that other phytochemical factors might contribute to the nephrotoxicity of the plant. In this study, the cytotoxicity in renal tubular cells (LLC-PK1) was measured using an aqueous extract from the plant and fractions from chromatographic separation to identify the cytotoxic constituents of the extract. In an iterative process two different groups of compounds were identified as the major cytotoxic principles in LLC-PK1 cells: steroidal saponins (primarily di- and trisaccharides of sarsasapogenin) and 5-hydroxy-4-methoxyfuran-2(5H)-one. Up to a concentration of 880 microg/mL (7.7 M) 3-methoxyfuran-2(5H)-one was not cytotoxic. The cytotoxicity of the saponins was abolished upon hydrolysis, indicating that the carbohydrate moiety of the molecule is a prerequisite for toxicity on the cellular level. The results of the present study have two important implications: first, the results question the direct involvement of 3-methoxyfuran-2(5H)-one in the nephrotoxicity of N. ossifragum; second, the findings should induce future investigations into the possible role of saponins in N. ossifragum-related nephrotoxicosis observed in ruminants that graze on this plant.     

Influence of abiotic stress on the GSH/GSSG system of plant cell cultures

Cell suspension cultures of various crop plants have been used for studying salt and drought stress on the GSH/GSSG-system, As already recorded for whole plants, cell cultures of different species vary in their glutathione levels. In general, cell cultures of species with relatively high glutathione content are more tolerant to abiotic stress. A high glutathione content alone is not adequate to prevent cell damage and a decrease in cell growth. E.g. the addition of sulphur to stressed cell cultures slightly enhances the GSH content but does not enhance the adaptation of these cells to stress. Thus, the role of glutathione in the adaptation of cells to stress situations seems to be only one component in a more complex response mechanism.     

Influence of delphinidin-3-glucoside on oxidized low-density lipoprotein-induced oxidative stress and apoptosis in cultured endothelial cells

Increased oxidative stress and apoptosis were detected in atherosclerotic lesions. Oxidized low-density lipoprotein (oLDL) may induce oxidative stress and apoptosis via multiple pathways in vascular endothelial cells (EC). Delphinidin-3-glucoside (D3G), an anthocyanidin glycan enriched in dark-skin berries, may neutralize those effects of oLDL in EC. The present study demonstrated that oLDL increased the generation of intracellular NADPH-dependent superoxide and impaired redox status in cultured porcine aortic EC (PAEC). The activities of mitochondrial respiratory chain complex I-IV and the contents of NADH dehydrogenase (ND)1, ND6 (complex I enzyme subunits), or cytochrome b (complex III enzyme subunit) were significantly reduced in PAEC treated with oLDL compared to controls. Treatment with oLDL significantly increased the abundances of NADPH oxidase (NOX)2, NOX4, and p22phox in PAEC. oLDL reduced cell viability and the protein content of B-cell lymphoma (Bcl)-2, but increased the content of caspase 3 in PAEC. Co-treatment with D3G prevented oLDL-induced increases in intracellular superoxide or in the protein content of NOX2, NOX4, p22phox, or caspase 3, inhibited the impairment of redox statues or cell viability, and prevented the attenuation of mitochondrial enzyme activities and the reductions of Bcl-2, ND1, or cytochrome b contents in PAEC. The findings suggest that oLDL induced oxidative stress and apoptosis in EC, which was associated with the activation of NOX, the impairment of mitochondrial respiration chain enzymes, and the disorder of key regulators for apoptosis. D3G neutralized the harmful effects of oLDL on oxidative stress, mitochondrial dysfunction, and apoptosis in cultured vascular EC.     

外源 GSH 对 NaCl 胁迫下番茄幼苗叶片及根系离子微域分布的影响

【目的】盐胁迫是限制新疆番茄生长的重要障碍因子之一,而外源喷施谷胱甘肽 (GSH) 是解决这一问题的有效措施。探讨外源 GSH 缓解番茄盐胁迫的效应和作用机制,可为该措施的有效应用提供理论依据。 【方法】采用营养液栽培法,选用番茄品种‘中蔬四号’为试材。在营养液中加入 NaCl 100 mg/L,使其产生盐胁迫,以不加 NaCl 作为对照 (CK),试验处理包括不喷施 GSH (NaCl)、喷施 GSH (+ GSH)、喷施 GSH 合成酶抑制剂 (+ BSO) 以及喷施 GSH 和 BSO (+ BSO + GSH)。测定番茄幼苗叶片和根系中与耐盐性相关的 K+、Ca2+、Mg2+、Na+ 和 Cl– 的离子微域分布状态和平衡。 【结果】NaCl 胁迫下番茄叶片和根系所有组织细胞中 Na+ 和 Cl– 相对含量显著提高,K+ 相对含量和 K+/Na+、Ca2+/Na+、K+/Cl– 比值降低,说明 NaCl 胁迫使细胞中 Na+ 和 Cl– 有害离子积累及胞内离子稳态严重破坏;外源 BSO 施用进一步加剧了 NaCl 胁迫下番茄叶片和根系细胞的 K+/Na+ 失衡。而外源 GSH 施用抑制了 NaCl 胁迫下番茄叶片和根系对 Na+ 的吸收,降低了 Cl– 的相对含量,提高了 K+/Na+、Ca2+/Na+、K+/Cl– 比值。外源 GSH 亦使 NaCl+BSO 胁迫下番茄叶片各组织及根系中皮层、内皮层和中柱的 Na+ 未检出,根系和叶片各组织中 Cl– 相对含量显著降低,K+ 和 Ca2+ 相对含量及 K+/Na+、Ca2+/Na+、K+/Cl–、Ca2+/Cl– 比值显著提高。 【结论】外源 GSH 通过抑制盐胁迫下番茄叶片和根系对 Na+ 的吸收,降低 Cl– 吸收,改善细胞中离子的微域分布和维持离子平衡, 从而缓解了盐胁迫对番茄的毒害作用,提高了番茄的耐盐性。