亚硒酸钠诱导SW480细胞凋亡过程中活性氧的作用

为探讨亚硒酸钠诱导人结肠癌SW480细胞凋亡的机理,将荧光探针2′,7′－二氯荧光黄乙二脂（2′,7′－DCFH－DA）、罗丹明123（rhodamine123）负载人结肠癌细胞,利用多光子成像系统测定胞内活性氧（ROS）、线粒体跨膜电位（△Ψm）的变化。结果发现（1）Na2SeO3作用SW480细胞,可导致细胞凋亡和胞内的ROS增加。SOD、过氧化氢酶可降低凋亡率并抑制ROS的增加。（2）线粒体电子传递链抑制剂鲁藤酮及氰化钠可抑制OS增加。（3）Na2SeO3可导致线粒体的跨膜电位的下降。表明Na2SeO3作用细胞可导致来源于线粒体的ROS增加,ROS介导亚硒酸钠诱导细胞凋亡。     

The antimutagenic effect of selenium on 7,12-dimethylbenz(a)anthracene and metabolites in the amesSalmonella/microsome system

The antimutagenic effect of selenium as sodium selenite, sodium selenate, selenium dioxide, and seleno-methionine was studied in the AmesSalmonella/microsome mutagenicity test using 7,12-dimethylbenz(a)anthracene (DMBA) and some of its metabolites. Selenium (20 ppm) as sodium selenite reduced the number of histidine revertants on plates containing up to 100 μg DMBA/plate. Increasing concentrations of selenium as sodium selenite, sodium selenate, and selenium dioxide up to 40 ppm Se progressively decreased the number of revertants caused by 50 μg DMBA. DMBA and its metabolites 7-hydroxymethyl-12-methylbenz(a)anthracene, 12-hydroxymethyl-7-methylbenz(a)anthracene, and 3-hydroxy-7,12-dimethylbenz(a)anthracene were mutagenic forSalmonella typhimurium TA100 in the presence of an S-9 mixture. Selenium supplementation as Na₂SeO₃ reduced the number of revertants induced by these metabolites to background levels. The antimutagenic effect of inorganic selenium compounds cannot be explained by toxicity of selenium as determined by viability tests withSalmonella typhimurium TA100. Selenium supplementation in all forms examined, except sodium selenate, decreased the rate of spontaneous reversion. Selenium as sodium selenate was slightly mutagenic at concentrations of 4 ppm or less. Higher concentration of Na₂SeO₄ inhibited the mutagenicity of DMBA. The present studies support the anticarcinogenic potential of selenium and indicate that form and concentration are important factors in this trace element's efficacy.     

Supplementation with organic or inorganic selenium in heat-distressed quail

The present study was carried out to determine the effects of different sources of selenium (Se; sodium selenite [Na₂SeO₃] or selenomethionine [Se-Met]) supplementation on egg production, egg quality, levels of malondialdehyde (MDA), and Se in serum and egg yolk in heat-stressed Japanese quail (Coturnix coturnix japonica). The birds (n = 360; 55 days old) were randomly assigned to 12 treatment groups consisting of six replicates of five birds each in a 2 × 2 × 3 factorial arrangement of treatments (temperatures, selenium sources, selenium levels). Birds were kept in wire cages in a temperature-controlled room at either 22 (thermoneutral) or 34°C (heat stress) for 8 h/day (09:00–17:00; till the end of study) and fed a basal (control) diet or the basal diet supplemented with either 0.15 or 0.30 mg of Na₂SeO₃ or selenomethionine/kg of diet. Heat exposure decreased live weight, feed intake, feed efficiency, egg production, and egg quality when basal diet was fed (P < 0.0001). A linear increase in feed intake (P = 0.001) and body weight (P = 0.001), egg production (P = 0.001), and improvement in feed efficiency (P = 0.001) and egg quality (P = 0.001) were found in Se-supplemented quail reared under heat stress conditions. Serum, egg white, and egg yolk Se (P ≤ 0.001) concentrations increased linearly, whereas serum, liver, and egg yolk MDA concentrations decreased linearly (P = 0.001) as dietary Na₂SeO₃ or Se-Met supplementation increased. An interaction between dietary Se sources, temperature, and levels of supplementation (P ≤ 0.05) for these parameters was detected. Supplementation with Se improved egg production, egg quality, and antioxidant status of birds, and the effects of Se-Met were relatively greater than Na₂SeO₃ in heat-stressed quail. Results of the present study suggest that supplementation with Se-Met can be considered to be more protective than Na₂SeO₃ by reducing the negative effects of oxidative stress induced by heat stress in quail.     

Arsenite-Induced Apoptosis Is Prevented by Selenite in A375 Cell Line

Arsenic trioxide induces apoptosis and clinical remission in patients diagnosed with acute promyelocytic leukemia. The human malignant melanoma A375 cells were treated with NaAsO₂ (0.1–130 μM) and also treated with combined 10 μM NaAsO₂ and 10 μM Na₂SeO₃. NaAsO₂ arrested cell growth in the G1 phase and induced apoptosis in a concentration- and time-dependent manner. In contrast, administration of Na₂SeO₃ antagonized the cell growth inhibition and apoptosis induced by NaAsO₂. The NaAsO₂ treatment resulted in a marked increase in p53 protein as early as 4 h and in Bcl-2 protein level by 12 h. In addition, p53 downregulation accompanied the combined treatment of NaAsO₂ and Na₂SeO₃. Thus, our results indicate upregulation of p53 and Bcl-2 play a crucial role in the NaAsO₂-induced G1 arrest and apoptosis of A375 cells and that downregulation p53 appears to contribute to the inhibition by Na₂SeO₃ of the effects induced by NaAsO₂.     

Hyperglycemia induces apoptosis and p53 mobilization to mitochondria in RINm5F cells

The mechanisms related to hyperglycemia-induced pancreatic β-cell apoptosis are poorly defined. Rat insulin-producing cells (RINm5F) cultured in high glucose concentrations (30 mM) showed increased apoptosis and protein p53 translocation to mitochondria. In addition, hyperglycemia induced both the disruption of mitochondrial membrane potential (Δ < eqid1 > _m), and an increase in reactive oxygen species (ROS), as shown by fluorescence changes of JC-1 and dichlorodihydrofluorescein-diacetate (DCDHF-DA), respectively. The increased intracellular ROS by high glucose exposure was blunted by mitochondrial-function and NADPH-oxidase inhibitors. We postulate that the concomitant mobilization of p53 protein to the mitochondria and the subsequent changes on the Δ < eqid2 > _m, lead to an important pancreatic β-cell apoptosis mechanism induced by oxidative stress caused by hyperglycemia. This work is part of the thesis required for the doctorate degree in Biological Sciences at the Universidad Autónoma Metropolitana, Mexico City, Mexico.     

Comparative assessment of selenite (SeIV) detoxification to elemental selenium (Se0) by <Emphasis Type="Italic">Bacillus</Emphasis> sp.

Bacillus licheniformis, B. subtilis, B. cereus, Bacillus pumilus and Exiguobacterium sp., which were resistant up to 20 mg Na₂SeO₃/ml in nutrient broth and 40 mg/ml on nutrient agar plates, were isolated from contaminated soil and water. They grew from 25 to 45°C and pH 5 to 9. They had multiple metal and antibiotic resistances. All strains reduced selenite (SeIV) to elemental selenium (Se0) aerobically with a maximum reduction of 97% by B. pumilus after 144 h with Na₂SeO₃ at 500 μg/ml.     

Enhancement of the antioxidants ergothioneine and selenium in Pleurotus eryngii var. eryngii basidiomata through cultural practices

Antioxidants are molecules that may reverse, prevent or slow cellular damage caused by free radicals. Increasing dietary intake of antioxidants is thought to reduce oxidative stress that may contribute to the development of several diseases. Mushrooms are known to contain antioxidants such as selenium, ergothioneine and phenolics that may serve this role. Here we sought to enhance selenium and ergothioneine concentration in Pleurotus eryngii var. eryngii basidiomata by modifying the techniques used for their commercial cultivation. To enhance selenium content in mushrooms, substrates were supplemented with sodium selenite (Na₂SeO₃) to reach selenium concentrations of 5 and 10 μg/g. Basidiomata of one commercial isolate (WC888) accumulated selenium up to 4.6 and 9.3 μg/g (d.w.), respectively. Therefore, a serving size (85 g) of fresh P. eryngii mushrooms produced on substrates supplemented with 5 and 10 μg/g of Na₂SeO₃ would supply 70.4 and 116.3% of the daily value of selenium (DV = 70 μg), respectively. Since selenium-enriched mushrooms would supply more than 20% of the DV, they could be considered an excellent source of selenium. Ergothioneine concentration was enhanced in mushrooms produced on low-moisture (55%) substrate compared to the commonly used 60% (high-moisture) in commercial cultivation. Mushrooms produced on low-moisture substrate had ergothioneine concentrations of 3.0 mg/g, while mushrooms produced on high-moisture substrate contained 2.2 mg/g or less. Use of a casing overlay for mushroom production resulted in significant yield increases on low-moisture substrate but not on high-moisture substrate.     

Selenium-enriched Candida utilis: Efficient preparation with l-methionine and antioxidant capacity in rats

Selenium-enriched Candida utilis has attracted much attention due to its expanding application in food and feed additives. The objective of this study was to efficiently prepare selenium-enriched C. utilis and to investigate the effects of the prepared yeast on antioxidant capacity in rats. A batch culture of selenium-enriched C. utilis was first carried out, and the addition of sodium selenite (Na₂SeO₃) after all glucose had been consumed was found to favor higher intracellular glutathione and organic selenium content. Moreover, l-methionine boosted yeast cell growth and glutathione biosynthesis, and prevented glutathione from leaking to the extracellular space that can be caused by Na₂SeO₃. We therefore developed a two-stage culture strategy involving supplementation with l-methionine and Na₂SeO₃ at separate culture phases to improve the performance of selenized C. utilis. Using this two-stage culture strategy, intracellular glutathione content reached 18.6 mg/g and 15.5 mg/g, respectively, in batch and fed-batch systems, and organic selenium content reached 905.2 μg/g and 984.7 μg/g, respectively. The effects of selenium-enriched C. utilis on the activities of antioxidant related enzymes in rats were investigated, and the prepared selenium-enriched C. utilis was shown to be an optimal dietary supplement for enhancing antioxidant capacity in rats.     

Molecular mechanisms of euplotin C-induced apoptosis: involvement of mitochondrial dysfunction,oxidative stress and proteases

The metabolite euplotin C (EC), isolated from the marine ciliate Euplotes crassus, is a powerful cytotoxic and pro-apoptotic agent in tumour cell lines. For instance, EC induces the rapid depletion of ryanodine Ca²⁺ stores, the release of cytochrome c from the mitochondria, and the activation of caspase-3, leading to apoptosis. The purpose of this study was to gain further insight into the mechanisms of EC-induced apoptosis in rat pheochromocytoma PC12 cells. We found that EC increases Bax/Bcl-2 ratio and that Bax is responsible of the EC-induced dissipation of the mitochondrial membrane potential (Δψ_m). In addition, EC induces the generation of reactive oxygene species (ROS) without involvement of p53. The inhibition of ROS generation prevents, at least in part, the pro-apoptotic effects of EC as well as the effects of EC on Bax, Δψ_m and intracellular free Ca²⁺, indicating a cross-talk between different pathways. However, definition of the effector cascade turns out to be more complex than expected and caspase-independent mechanisms, acting in parallel with caspases, should also be considered. Among them, EC increases the expression/activity of calpains downstream of ROS generation, although calpains seem to exert protective effects. D. Cervia and M. Garcia-Gil equally contributed to the work.     

Homocysteine affects cardiomyocyte viability: concentration-dependent effects on reversible flip-flop, apoptosis and necrosis

Background Hyperhomocysteinaemia (HHC) is thought to be a risk factor for cardiovascular disease including heart failure. While numerous studies have analyzed the role of homocysteine (Hcy) in the vasculature, only a few studies investigated the role of Hcy in the heart. Therefore we have analyzed the effects of Hcy on isolated cardiomyocytes. Methods H9c2 cells (rat cardiomyoblast cells) and adult rat cardiomyocytes were incubated with Hcy and were analyzed for cell viability. Furthermore, we determined the effects of Hcy on intracellular mediators related to cell viability in cardiomyocytes, namely NOX2, reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨ _m) and ATP concentrations. Results We found that incubation of H9c2 cells with 0.1 mM D,L-Hcy (= 60 μM l-Hcy) resulted in an increase of ΔΨ _m as well as ATP concentrations. 1.1 mM d,l-Hcy (= 460 μM l-Hcy) induced reversible flip-flop of the plasma membrane phospholipids, but not apoptosis. Incubation with 2.73 mM d,l-Hcy (= 1.18 mM l-Hcy) induced apoptosis and necrosis. This loss of cell viability was accompanied by a thread-to-grain transition of the mitochondrial reticulum, ATP depletion and nuclear NOX2 expression coinciding with ROS production as evident from the presence of nitrotyrosin residues. Notably, only at this concentration we found a significant increase in S-adenosylhomocysteine which is considered the primary culprit in HHC. Conclusion We found concentration-dependent effects of Hcy in cardiomyocytes, varying from induction of reversible flip-flop of the plasma membrane phospholipids, to apoptosis and necrosis.     

In vitro differential effects of sodium selenite on the growth of human hepatoma cells and human embryonic liver cells

The effects of various concentrations of Na₂SeO₃ on human hepatoma cells and human embryonic liver cells was investigated in vitro. For human hepatoma cells, mitotic index and cell count decreased with increasing selenium concentrations. At 1 μg/mL Na₂SeO₃, mitotic activity of human hepatoma cells were partially arrested. In human embryonic liver cells continuously treated with Na₂SeO₃, (1 μg/mL) cell count of the treated group decreased only by d 7; mitotic index, labeled index, and mean silver grain number per 50 labeled nuclei were the same as in the control group on exposure to 1, 3, and 5 μg/mL for up to 72 h. In mixed cultures of human hepatoma and embryonic liver cells treated with 3 and 5 μg/mL of Na₂SeO₃ for 24 h, hepatoma cells showed vacuolated cytoplasms, distorted nuclei, condensed chromatin, and even pyknosis, whereas the embryonic liver cells retained a normal morphology under the same treatment.     

Difference in Selenite Absorption Between High- and Low-Selenium Rice Cultivars and its Mechanism

Two rice cultivars, Xiushui 48 and S. Andrea, with significant difference in selenium (Se) concentrations in brown rice grains, were chosen to study the Se absorption and its mechanism in excised roots. The results showed that the high-selenium cultivar Xiushui 48 absorbed higher amounts of Na₂SeO₃ than low-selenium S. Andrea at different Se levels and treatment periods. It was found that Na₂SO₃ markedly inhibited Na₂SeO₃ absorption by the excised roots of both cultivars. This inhibition might be due to the competition for uptake on the fact that Na₂SO₃ might share a common uptake pathway with Na₂SeO₃. Treatment with ZnCl₂ significantly decreased Na₂SeO₃ absorption of both cultivars possibly by inhibiting the activity of cysteine synthase. It was therefore postulated that the difference in cysteine synthase activity might be one of the reasons which resulted in difference in selenite absorption possibly between the two cultivars. Both HgCl₂ and AgNO₃ treatments can inhibit selenite absorption by rice roots greatly. We propose that selenite enters rice roots through aquaporins as the form of H₂SeO₃.     

Growth and biochemical alterations in coffee due to selenite toxicity

Two experiments were conducted to investigate selenite toxicity in coffee (Coffea arabica cv. Catuaí). In the first aqueous selenite solution (10 µM Na₂SeO₃) was used to infiltrate leaves of an adult coffee plant. The infiltrated leaves and fruits adjacent to them showed enhanced contents of caffeine and soluble sugars. Amino acid contents were not affected, whereas pigments (chlorophylls, carotenoids and xanthophylls) exhibited a significant decrease. In the second experiment, coffee seedlings were irrigated with aqueous selenite solutions (10,100 and 1000 µM Na₂SeO₃) and the first and third pairs of leaves were analyzed. Control plants did not receive selenium. The plants were not different in height, but at the highest selenium concentration showed lower dry matter accumulation in roots and leaves, lower leaf area and thicker leaves. Increases in caffeine and soluble sugars were observed in the first pair of leaves at the highest selenium concentration, although selenium content itself increased steadily with increasing solution concentration. Phenols increased in both leaf pairs and pigments decreased in the third pair. Nitrate reductase activity, measured in the second leaf pair, was much lower at all selenium levels. The profile of free amino acid was altered in leaves of plants treated with selenium.     

The influence of selenium and caffeine on chemical carcinogenesis in rats,mutagenesis in bacteria,and unscheduled DNA synthesis in human lymphocytes

The influence of sodium selenite (Na₂SeO₃) and caffeine on chemical carcinogenesis induced in rats by diethylnitrosamine (DEN), N-nitrosomorpholine (NM), andN-methyl-N-nitro-N-nitrosoguanidine (MNNG) was investigated. A dose-dependent inhibitory effect of Na₂SeO₃ (l–10 ppm) on hepatocarcinogenesis induced by DEN was demonstrated. Na₂SeO₃ also increased the latency period for stomach tumor formation in rats treated with MNNG. Combined treatment of rats with Na₂SeO₃ plus vitamin C added to the diet resulted in a slight inhibition of NM-induced liver carcinogenesis. Supplementation of diet with Na₂SeO₃ plus butylated hydroxytoluene, vitamin C, and vitamin E did not reveal any additive inhibitory effect compared to the inhibitory effect of Na₂SeO₃ given alone. Caffeine (600 rag/L) reduced the number of liver tumors induced in rats by DEN. Preliminary experiments have indicated that combined treatment of rats with selenium and caffeine could result in more effective inhibition of DEN-induced liver carcinogenesis. Further experiments are being conducted to study the influence of selenium and caffeine on mutagenic activity of 1-methyl-l-nitrosourea (MNU) inSalmonella typhimurium TA 1535. The pretreatment of bacteria cells with Na₂SeO₃ (3–10 p.g/mL) increased the mutagenic response of bacteria to MNU. A synergistic stimulation of mutagenic activity of MNU was observed in bacteria pretreated simultaneously with Na₂SeO₃ and caffeine. In addition the influence of Na₂SeO₃ on UDS induced by DEN in human lymphocytes was investigated. The trace element inhibited the UDS up to 82%. The possible role of potentiation by NazSeO3 of the cell killing effect of DEN in inhibition of liver carcinogenesis was discussed.     

Differential Effects of selenium on the proliferation of human pulmonary adenocarcinoma cells and human embryonic lung diploid cells in vitro

The effect of different concentrations of Na₂SeO₃ on human pulmonary adenocarcinoma cells and human embryonic lung diploid cells in vitro was investigated. For human pulmonary adenocarcinoma cells, mitotic index and cell count decreased with increasing selenium concentrations. At 1 μg/mL sodium selenite, mitotic activity and growth of human lung cancer cells were partially inhibited, and the progression of human lung cancer cell cycle was partially arrested. When human embryonic lung diploid cells were treated with 1 μg/mL sodium selenite for five continuous days, cell counts of the treated group were closely parallel to those of the control group. After treating human embryonic lung diploid cells with 1–5μg/mL sodium selenite for 1–3 d, the mitotic index (MI), labeled index (LI), and average silver grain (SG) number per 20 labeled nuclei were the same as those of the control. In mixed cultures of human embryonic lung diploid cells and human pulmonary adenocarcinoma cells, treated with 3 and 5 μg/mL sodium selenite for 24 h, the lung diploid cells showed a normal fusiform morphology, whereas the lung cancer cells showed heavily vacuolated cytoplasms and distorted nuclei.     

Cytotoxicity of sodium selenite in HaCaT cells induces cell death and alters the mRNA expression of PUMA,ATR, and mTOR genes

BackgroundBy identifying the molecular mechanisms underlying sodium selenite (Na₂SeO₃) cytotoxicity during exposure in non-tumor cells (HaCaT cells), we will improve the current understanding of its antiproliferative effects and modulation of gene expression in the main pathways related to the cell cycle, cell death, oxidative stress, and DNA damage and repair.MethodsNon-tumor HaCaT cells were treated with Na₂SeO₃ to induce cytotoxicity, and the effects were investigated using an MTT assay (cell viability), real-time cell analysis (profiling the cell index), flow cytometry (membrane integrity, cell cycle disruption, and apoptosis), a comet assay (genotoxicity, i.e., DNA damage), and RT-qPCR (mRNA expression of genes).ResultsTreatment with Na₂SeO₃ was cytotoxic at 10 μM, producing morphological changes in cells (cytoplasmic granulations); however, it did not have a genotoxic effect. Na₂SeO₃ induced cell membrane damage, cell death, and cell cycle arrest in HaCaT cells. It also altered the mRNA expression levels of PUMA, ATR, and mTOR genes. However, it had no effect on the mRNA expression of caspases or PARP1, BIRC5, BECN1, and c-MYC genes, suggesting that Na₂SeO₃ causes PUMA-dependent apoptosis in HaCaT cells. The mRNA expression of specific genes related to oxidative stress, DNA damage and repair, and cell cycle control were unchanged by Na₂SeO₃.ConclusionsWe demonstrated the cytotoxic effect of Na₂SeO₃ in HaCaT cells by analyzing mRNA expression patterns, changes in cell morphology, and proliferation kinetics.     

Anti-Cancer Effect and the Underlying Mechanisms of Gypenosides on Human Colorectal Cancer SW-480 Cells

Background

Gypenosides (Gyp), the main components from Gynostemma pentaphyllum Makino, are widely used in traditional Chinese medicine. The present study aimed to investigate the anti-cancer effect and the underlying mechanisms of Gyp on human colorectal cancer cells SW-480.

Materials and Methods

The inhibitory effect of Gyp on SW-480 cells was evaluated by MTT assay. Apoptotic cell death was detected by nuclear Hoechst 33342 staining and DNA fragmentation analysis. Apoptosis was analyzed using Annexin V-PE/7-amino-actinomycin D staining. Cell membrane integrity was evaluated with flow cytometry following PI staining. Changes of mitochondrial membrane potential (Δψ _m) were detected through flow cytometry analysis of rhodamine 123 (Rh123). The role of reactive oxygen species (ROS) in Gyp induced cell death was investigated by intracellular ROS generation and general ROS scavenger. Wound-healing assay was carried out to investigate Gyp-inhibited migration of SW-480 cells in vitro. Additionally, the alterations in F-actin microfilaments were analyzed by FITC-labeled phalloidin toxin staining and the morphological changes were evaluated under scanning electron microscope (SEM).

Results

After the Gyp treatment, the plasma membrane permeability of SW-480 cell was increased, Δψ _m was decreased significantly, the level of intracellular ROS level was increased, DNA fragmentation and apoptotic morphology were observed. Cells treated with Gyp exert serious microfilament network collapse as well as the significant decrease in the number of microvilli. Gyp induced the changes of cell viability, cell migration, intracellular ROS generation and nuclear morphology were alleviated obviously by NAC.

Conclusion

The results in this study implied that ROS play an important role in Gyp induced cell toxicity and apoptosis, and the mitochondria damage may be upstream of ROS generation post Gyp treatment. The findings of the present study provide new evidences for anti-tumor mechanisms by which Gyp induces apoptosis in vitro.     

Effects of Sodium Selenite and Germination on the Sprouting of Chickpeas (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and Its Content of Selenium,Formononetin and Biochanin A in the Sprouts

To improve the nutritional value of chickpea food, selenium (Se)-rich chickpea sprouts were produced by germination of chickpea seeds for 6 days at 28 centigrade in the presence of various concentrations of Na₂SeO₃ in germination solution. High concentrations of selenite were found to inhibit the growth of chickpea sprout and the biosynthesis of isoflavones formononetin and biochanin A. However, chickpea sprouts could tolerate up to ∼50 mg/L of Na₂SeO₃, under which condition the product chickpea sprouts contained a high Se content (2.14 μg/g dry weight) and a moderate high content of isoflavones (601.56 μg biochanin A/g dry weight and 578.11 μg formononetin/g dry weight). Se was incorporated in chickpea sprout in the form of selenomethionine. Thus, Se-enriched chickpea sprouts may serve as a convenient dietary source of Se and of isoflavones, including formononetin and biochanin A.     

Anti-hemolytic and Peroxyl Radical Scavenging Activity of Organoselenium Compounds: An In Vitro Study

Selenium-containing amino acids, selenocystine (CysSeSeCys), methylselenocysteine (MeSeCys), and selenomethionine (SeMet) have been examined for anti-hemolytic and peroxyl radical scavenging ability. Effect of these compounds on membrane lipid peroxidation, release of hemoglobin, and loss of intracellular K⁺ ion as a consequence of peroxyl radicals-induced oxidation of human red blood cells were used to evaluate their anti-hemolytic ability. The peroxyl radicals were generated from thermal degradation of 2,2′-azobis(2-methylpropionamidine) dihydrochloride. Significant delay (t _eff) was observed in oxidative damage in the presence of the selenium compounds. From the IC₅₀ values for the inhibition of hemolysis, lipid peroxidation, and K⁺ ion leakage, the relative anti-hemolytic ability of the compounds were found to be in the order of CysSeSeCys > MeSeCys > SeMet. The anti-hemolytic abilities of the compounds, when compared with sodium selenite (Na₂SeO₃) under identical experimental conditions, were found to be better than Na₂SeO₃. Relative rate constants estimated for the reaction of MeSeCys and SeMet with peroxyl radicals by competition kinetics using ABTS²⁻ as a reference confirmed that all the compounds are efficient peroxyl radical scavengers. Comparison of the GPx-like activity of these compounds, by NADPH–GSH reductase coupled assay, indicated that CysSeSeCys exhibits the highest activity. Based on these results, it is concluded that among the compounds examined, CysSeSeCys, possessing the ability to reduce peroxyl radicals and hydroperoxides showed efficient anti-hemolytic activity.