Hydrogen Peroxide-induced Hemolysis Research Articles

Chemical characterization and in vitro biological evaluation of aqueous extract of Althaea officinalis L. flower grown in Lebanon

IntroductionHerbal medicine is extensively used in various therapeutic strategies. Althaea officinalis L. of the family Malvaceae exhibits many biological activities through its phenolic acids, flavonoids, coumarins and polysaccharides. In Lebanon, the flowers are consumed in hot infusions. Therefore, the current study aimed to evaluate chemical and biological properties of Lebanese Althaea officinalis L. flower water extract. MethodsChemical characterization was performed by qualitative phytochemical screening, gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-mass spectrometry (LC-MS). Anti-proliferative effect was evaluated in cancer cells by MTT assay. The mRNA expression levels of pro-inflammatory enzyme and inflammatory cytokines were assessed in LPS-stimulated RAW 264.7 cells. Cytoprotective activity was examined in red blood cells against hydrogen peroxide-induced hemolysis. Antioxidant property was assessed by DPPH radical scavenging assay. ResultsQualitative phytochemical screening of the extract revealed the presence of anthraquinones, flavonoids, lignins, phenols, tannins, terpenoids and others. GC/MS analysis suggested that most of the phytocompounds are conjugated to sugars rather than free. LC/MS analysis identified 5 phenolic acids (syringic, gallic, caffeic, p-coumaric and trans-ferulic acids) and 8 flavonoids (catechin, apigenin, chrysin, quercetin, kaempferol, genistein, rutin trihydrate and galangin). Biological evaluation of the extract showed anti-proliferative effects on A549, EB, HCT-116, MCF-7 and HeLa 229 cells, anti-inflammatory activity illustrated by decrease in mRNA expression of inducible nitric oxide synthase, interleukin 1 beta, tumor necrosis factor alpha and interleukin 6, cytoprotective activity in red blood cells and antioxidant property. ConclusionAlthaea officinalis flowers rich in phytocompounds exert interesting biological activities and hold promise in the pharmacological field.

Metabolite profiling, anti-inflammatory, analgesic potentials of edible herb Colocasia gigantea and molecular docking study against COX-II enzyme

Ethnopharmacological relevanceConsumable herbs play a basic part in sustenance and human health. Traditionally, Colocasia gigantea Hook (Araceae) is used to treat fever, infection, wounds healing, drowsiness, tuberculosis, stomach problems etc. Aim of the studyThe study aspired to identify bioactive compounds, to evaluate anti-inflammatory and analgesic potentials of edible herb C. gigantea, and to molecular docking study against anti-inflammatory enzyme Cyclooxygenase-2 (COX-2). Materials and methodsChemical components of C. gigantea were discerned by HPLC and GCMS assays. In vitro anti-inflammatory activity was appraised by heat-induced, hypotonicity, and hydrogen peroxide-induced hemolysis assays and in vivo by formalin-induced paw edema assay. In vivo analgesic activity was evaluated by acetic acid-induced pain modulation assay. Also, molecular docking of the identified compounds was explored against the anti-inflammatory enzyme cyclooxygenase-2. ResultsHPLC-DAD analysis divulged the presence of trans-cinnamic acid along with (−)-epicatechin as a prime component. Also, 9, 12-Octadecadienoic acid (37.86%) and n-Hexadecanoic acid (25.89%) as the major as well as 24 other compounds were confirmed through GCMS in the extract. In in vitro anti-inflammatory study, C. gigantea extract indicated prominent erythrocyte membrane stabilization activity with good percentage aegis in all experimental assays. In addition to, formalin-induced in vivo anti-inflammatory assay revealed the maximum (42.37% and 48.72%) suppression of edema at the fourth hour at 250 and 500 mg/kg body weight, respectively. Moreover, an in-vivo pain modulation assay exposed significant (p < 0.05) activity at experimental doses. Furthermore, in the docking study, (−)-epicatechin was more active rather than other identified compounds with strong binding affinity to COX-2 protein. ConclusionsThe extract evinced remarkable anti-inflammatory and analgesic activities. Identified bioactive components along with other components of the extract might play a pivotal role in the observed bioactivity and the results vindicate the use of edible herb C. gigantea in ancestral medicine.

In-vitro Anti-lipid Peroxidation and Other Antioxidant Potentials of Gongronema latifolium Leaf Extract

Gongronema latifolium is a herbaceous plant consumed as a traditional folk medicine. The aim of this work is to evaluate the anti-lipid peroxidation activity and other antioxidant effects of the plant leaf extract of Gongronema latifolium using various in vitro models. Lipid peroxidation was first induced in egg yolk and bovine liver homogenate using the ascorbate-ferrous system and incubated with the plant extract at different concentrations. In another experiment, hydrogen peroxide was used to induce erythrocyte hemolysis and lipid peroxidation and those erythrocytes were incubated with the extract. Finally the potential ferrous reducing ability, hydroxyl radical and hydrogen peroxide scavenging activities of the plant extract were also analyzed. It revealed that the leaf extract significantly reduced chemically-induced lipid peroxidation in both homogenates when compared to quercetin. The extract also reduced both hydrogen peroxide-induced hemolysis and lipid peroxidation in erythrocytes. The extract also possessed significant abilities to reduce ferric ions, scavenge both hydroxyl radicals and hydrogen peroxide. In most cases, the responses were concentration-dependent (p &lt; 0.05). The findings are ascribed to the important phytochemicals which are antioxidant in nature hence the plant could be exploited both pharmacologically and neutraceutically.

Evaluation of In vitro antioxidant potential of active metabolite constituents of different extracts of Chaetomium cupreum-SS02 by spectrophotometric method

Objective: The main objective of the study was to evaluate the antioxidant activities of Chaetomium cupreum extracts. Materials and Methods: The total flavonoid content was determined by using aluminum chloride method, whereas antioxidant activity (AA) was evaluated by ferric reducing antioxidant power assay, potassium ferricyanide reducing power assay, 2,2-diphenyl-1-picyl-hydrazyl method, β-carotene bleaching assay, cupric ion reducing antioxidant capacity assay, lipid peroxidation inhibition assay by thiobarbituric acid (TBA)-reactive substance method, and inhibition of hydrogen peroxide-induced erythrocyte hemolysis assay. Results: The ferric reducing AA of C. cupreum extracts at the concentration of 50 μg/mL was higher in ethyl acetate extract (6.11%) followed by chloroform extract (3.96%), n-butanol extract (2.44%), and methanol extract (2.02%) mg RE/g dry weight. The potassium ferricyanide reducing activity of C. cupreum extracts at the concentration of 50 μg/mL was higher in ethyl acetate extract (15.90%) followed by chloroform (9.50%), n-butanol (4.93%), and methanol extract (2.92%). The 2, 2-diphenyl-1-picryl-hydrazyl activity of C. cupreum extracts at 50 μg/mL was higher in ethyl acetate extract (36.13%) followed by n-butanol extract (24.17%), chloroform extract (15.04%), and methanol extract (4.71%). The β-carotene bleaching activity of C. cupreum extracts at 50 μg/mL after 1 h of incubation was higher in ethyl acetate extract at 12.88%, followed by chloroform extract (9.82%), n-butanol extract (5.63%), and methanol extract (3.76%). The cupric ion reducing AA (CUPRAC) of C. cupreum extracts at 50 μg/mL was highest in the methanol extract (18.62%) followed by ethyl acetate extract (9.72%), n-butanol extract (7.18%), and chloroform extract (2.46%) mg ACE/g dry weight. With regard to TBA reactive substance activity of C. cupreum extracts at 50 μg/mL, n-butanol extract showed the highest lipid peroxidation inhibition (55.39%) followed by chloroform extract (50.51%), ethyl acetate extract (46.27%), and methanol extract (43.60%). With regard to the hydrogen peroxide-induced hemolysis inhibition activity of C. cupreum extracts at the concentration of 500 μg/mL, ethyl acetate extract showed the highest inhibition (30.53%) followed by chloroform extract (26.42%) and n-butanol extract (9.16%). Conclusion: The results of the present study showed that C. cupreum extracts poses significant antioxidant potential.

Antioxidant evaluation and inhibitory activity of Chromolaena odorata leaf extract on hydrogen peroxide-induced erythrocyte hemolysis

Aim: Chromolaena odorata has been used traditionally for the treatment of various conditions which include skin infections, wounds, ulcers and also used as an antimalarial. The present work investigates the ability of the methanolic extract of the plant to inhibit hydrogen-peroxide induced haemolysis and lipid peroxidation of human erythrocytes. Other in vitro antioxidant and free radical scavenging activities were also investigated. Materials and Methods: Dried leaves of Chromolaena odorata were pulverized and extracted using absolute methanol. Veinous blood was collected from a healthy individual and erythrocytes isolated. Erythrocyte haemolysis and lipid peroxidation were induced using hydrogen peroxide and subsequently treated with Chromolaena odorata extract. The plant extract was also assessed for its hydrogen peroxide scavenging, hydroxyl radical scavenging and reducing abilities. In all cases, vitamin C was used as the reference compound. Results: The results show that Chromolaena odorata extract reduced hydrogen peroxide-induced haemolysis and lipid peroxidation when compared to vitamin C. While the IC50 values for the extract were 4.11 mg/mL and 12.95 mg/mL for inhibition of erythrocyte haemolysis and lipid peroxidation respectively, the IC50 values for vitamin C were 1.98 mg/mL and 0.57 mg/mL for haemolysis and lipid peroxidation. The extract also possessed hydrogen peroxide scavenging, hydroxyl radical scavenging and reducing abilities. In most cases, the effects were concentration-dependent and significant among various extracts concentrations (p < 0.05). Conclusion: The observed responses indicate that the plant possessed immense antioxidant potential which is ascribed to the bioactive phytochemicals. This could be exploited pharmacologically.

Antioxidant and Antihemolytic Activities of Ethanolic Extract of Flowers, Leaves, and Stems of Hyssopus officinalis L. Var. angustifolius

In this study, antioxidant and antihemolytic activities of ethanolic extract of flowers, leaves, and stems of Hyssopus officinalis L. Var. angustifolius were investigated employing different in vitro assay systems. Extracts showed good antioxidant activity. IC50 for 1,1-diphenyl-2-picryl hydrazyl radical-scavenging activity were 148.8 ± 4.31 μg mL−1 for flowers, 79.9 ± 2.63 μg mL−1 for stems, and 208.2 ± 6.45 μg mL−1 for leaves. All extracts showed moderate iron (II) chelating ability. Extracts exhibited good antioxidant activity in the hemoglobin-induced linoleic acid model and also they were capable of scavenging hydrogen peroxide in a concentration dependent manner. Extracts showed good antihemolytic activity againts hydrogen peroxide-induced hemolysis (IC50 were 48.51 ± 2.27 μg mL−1 for flowers, 19.47 ± 0.73 μg mL−1 for leaves, and 63.1 ± 2.65 μg mL−1 for stems). The total amount of phenolic compounds in the extracts was determined as gallic acid equivalents and total flavonoid content was calculated as quercetin equivalents from a calibration curve.

Effects of Vitamin E and Vitamin C on Hydrogen Peroxide-Induced Hemolysis in Moroccan Dromedary Camels (Camelus Dromedarius)

Antioxidant vitamins may affect an organism’s capacity for defense against damage caused by reactive oxygen species (ROS), and biological markers of the dietary exposure to these compounds is of importance. The present study was designed to evaluate the protective effect of vitamin E and vitamin C against the hemolysis induced by hydrogen peroxide (H 2O2) in camel’s red blood cells in vitro, and for assessing the vulnerability of camel erythrocytes to oxidative stress (OS). Vitamin E and vitamin C were able to protect the red blood cells against the H2O2-induced hemolysis and thiobarbituric acid reactive species (TBARS) formation. The antioxidative property of these vitamins in reducing oxidative injury showed in this work, may suggest that these molecules may be very helpful to fight the ROS during OS induced by transportation, heat, deshydratation, aging, and parasite, infectious and metabolic diseases in dromedary camels.

Antioxidant and antihemolytic activity of lipid-soluble bioactive substances in avocado fruits

Introduction. Persea americana (avocado) fruit is known to be a rich source of proteins and minerals, as well as vitamins. Although many biological activities have been reported for avocado fruit, far less attention has been paid to the biological properties of its lipid-soluble bioactive substances. The aim of this study was to evaluate the antioxidant and antihemolytic activity of lipid-soluble bioactive compounds of avocado fruit. Materials and methods. Antioxidant and antihemolytic activity of lipid-soluble bioactive compounds of avocado fruit was assessed by employing different methods. Also, the ability of avocado fruit to protect fluorescein against oxidant-induced bleaching was evaluated. Results and discussion. The tested sample showed good antioxidant and antihemolytic activities. There was no significant difference between the reducing power of lipid-soluble bioactive substances in avocado fruits and ascorbic acid (p < 0.05). The tested sample showed weak ferrous ion-chelating activity. The sample exhibited moderate hydrogen peroxide scavenging activity, but exhibited good antioxidant activity. Avocado fruit showed very good activity against 2,2′-azobis(2-amidinopropane) dihydrochloride- and hydrogen peroxide-induced hemolysis in erythrocytes. Furthermore, it showed good protective effects against peroxyl radical-induced bleaching in fluorescein. Conclusion. Lipid-soluble bioactive substances of avocado fruit showed good activity in all of the studied models.

In Vitro Antioxidant and Antihemolytic Activities of Hydroalcoholic Extracts of Allium scabriscapum Boiss. & Ky. Aerial Parts and Bulbs

This study is designed to evaluate the antioxidant and antihaemolytic activities of the hydroalcoholic extracts of Allium scabriscapum aerial parts and bulbs by employing eight in vitro assay systems. In 1,1-diphenyl-2-picryl hydrazyl radical scavenging assay, both the extracts show moderate scavenging activity. The reducing power ability of extracts increased with increasing in the samples concentrations. IC50 for metal chelating activity of aerial parts and bulbs extracts were 894.6 ± 31.29 and 746.2 ± 26.11 μg mL−1, respectively. The aerial part extracts show better nitric oxide and hydrogen peroxide scavenging activities than the bulb. Extracts exhibited good antioxidant activity in linoleic acid emulsion system and were comparable to vitamin C (p > 0.05). Aerial parts extract showed better antihemolytic activity against cumene hydroperoxide and hydrogen peroxide-induced hemolysis. Among the extracts A. scabriscapum, aerial parts had higher phenolic and flavonoid contents.

Interaction of Different Extracts of Primula heterochroma Stapf. with Red Blood Cell Membrane Lipids and Proteins: Antioxidant and Antihemolytic Effects

ABSTRACTIn recent years, considerable attention has been paid to plants as potent natural drugs for their ameliorative roles against free-radical-mediated oxidative stress. Therefore, their interactions with cell membrane lipids and proteins, which generally serve as primary targets of lipid peroxidation, are of much interest. In the current investigation, in vitro and ex vivo studies are performed in order to estimate possible effects of different extracts of Primula heterochroma Stapf. on red blood cell membranes of rat erythrocytes using colorimetric methods. The results indicate that binding of the extracts to lipids and proteins of red blood cell membranes both significantly inhibits lipid peroxidation, and also increases red blood cell integrity against hemolysis. Moreover, a polyphenol extract, in particular, demonstrates notable antihemolytic activity in hydrogen peroxide-induced hemolysis model (IC50 = 199.49 ± 9.1 μg ml−1).

Antioxidant Activity and Protective effect of Banana Peel against Oxidative Hemolysis of Human Erythrocyte at Different Stages of Ripening

Phytochemicals such as polyphenols and carotenoids are gaining importance because of their contribution to human health and their multiple biological effects such as antioxidant, antimutagenic, anticarcinogenic, and cytoprotective activities and their therapeutic properties. Banana peel is a major by-product in pulp industry and it contains various bioactive compounds like polyphenols, carotenoids, and others. In the present study, effect of ripening, solvent polarity on the content of bioactive compounds of crude banana peel and the protective effect of peel extracts of unripe, ripe, and leaky ripe banana fruit on hydrogen peroxide-induced hemolysis and their antioxidant capacity were investigated. Banana (Musa paradisica) peel at different stages of ripening (unripe, ripe, leaky ripe) were treated with 70% acetone, which were partitioned in order of polarity with water, ethyl acetate, chloroform (CHCl₃), and hexane sequentially. The antioxidant activity of the samples was evaluated by the red cell hemolysis assay, free radical scavenging (1,1-diphenyl-2-picrylhydrazyl free radical elimination) and superoxide dismutase activities. The Folin-Ciocalteu's reagent assay was used to estimate the phenolic content of extracts. The findings of this investigation suggest that the unripe banana peel sample had higher antioxidant potency than ripe and leaky ripe. Further on fractionation, ethyl acetate and water soluble fractions of unripe peel displayed high antioxidant activity than CHCl₃ and hexane fraction, respectively. A positive correlation between free radical scavenging capacity and the content of phenolic compound were found in unripe, ripe, and leaky ripe stages of banana peel.

The potential effect of puerarin in preventing atherosclerosis

To study the activity of Puerarin (Pue) in scavenging oxygen free radical (OFR) and its inhibitory effect on the oxidative modification of low density lipoprotein (LDL). Riboflavin-light system was used to generate superoxide anion, and Fenton reaction to generate hydroxyl free radical to study the activity of Pue in scavenging OFR. Hydrogen peroxide-induced hemolysis was used to study the effect of Pue on erythrocyte hemolysis and malondialdehyde (MDA) production. And ultraviolet ray and cupric sulfate were used to cause the oxidative modification of LDL for studying the inhibitory effect of Pue on LDL oxidative modification. (1) Pue could, at concentration of 0. 01 – 1.0 mmol/L, scavenge superoxide anion radical and at concentration of 7.5–75μmol/L scavenge hydroxyl radical in a concentration dependent manner. (2) Pue could, at concentration of 0.1–10 mmol/L, inhibit significantly oxidative hemolysis and MDA production of erythrocyte induced by hydrogen peroxide. (3) Pue of 0.01–1.0 mmol/L could inhibit the oxidative modification of LDL in a concentration dependent manner. Pue has an anti-peroxidation effect and shows a potential effect in preventing atherosclerosis.

Vitamin E deficiency and erythrocyte deformability in the rat

It has been suggested that erythrocyte deformability is decreased in vitamin E deficiency due to oxidative damage to the cell membrane. Male Wistar rats (66 to 88 g) were fed ad libitum an AIN76-based diet containing tocopherol-stripped corn oil without added vitamin E for 8 weeks (−E; n = 8). Control animals were fed ad libitum the same diet containing 50 IU/kg dl-α-tocopheryl acetate (+E; n = 7). Vitamin E deficiency was confirmed by depressed mean (±SEM) plasma α-tocopherol levels (μmol/L), as measured by high performance liquid chromatography [−E: 0.5 ± 0.1; +E: 20.3 ± 1.8] and elevated hydrogen peroxide-induced hemolysis (%) [−E: 92.6 ± 2.4; +E: 4.2 ± 1.6; P < 0.05 by Student's t test]. The only alteration in a complete blood count was a depression in reticulocyte number (× 10 12/L) [−E: 0.19 ± 0.02; +E: 0.46 ± 0.03; P < 0.05]. Erythrocyte deformability was measured at standard shear stress under conditions of increasing osmolality in the ektacytometer. Elongation index (the ratio of length to width of the diffraction pattern of the deformed cells) was plotted against osmolality to generate an osmotic deformability profile. EI max (the maximum elongation index) and O hyper (the osmolality at which the elongation index is half of EI max on the hypertonic arm of the curve) were significantly increased in samples from the −E group ( P < 0.05 by Student's t test). In summary, erythrocyte deformability as measured by the ektacytometer was not decreased by a subclinical vitamin E deficiency in the rat. In fact, a small but significant increase in maximum deformability was observed in erythrocytes from vitamin E-deficient rats.

Plasma vitamins A and E and hydrogen peroxide-induced in vitro erythrocyte haemolysis in common marmosets (Callithrix jacchus).

Plasma total lipid, cholesterol, all-trans retinol (vitamin A) and alpha-tocopherol (vitamin E) concentrations, and the susceptibility of erythrocytes to hydrogen peroxide-induced haemolysis in vitro were investigated in healthy laboratory-bred common marmosets (Callithrix jacchus). The concentrations of alpha-tocopherol and total cholesterol were similar to those of control human subjects. The mean lipid concentration was higher (P less than 0.025) and the retinol concentration lower (P less than 0.001) in the marmosets. The susceptibility of the erythrocytes of the marmosets to hydrogen peroxide-induced haemolysis was high, both in absolute value and relative to the controls. Changes in phospholipid and fatty acid composition might have rendered the red cells susceptible to oxidative stress.

N-3 and N-6 phosphoglyceride fatty acids in relation to in vitro erythrocyte haemolysis induced by hydrogen peroxide in captive common marmosets ( Callithrix jacchus)

1. 1. Hydrogen peroxide-induced haemolysis (HPIH) was studied in the red blood cells of the common marmoset monkey ( Callithrix jacchus) in relation to the composition of the membrane fatty acids. HPIH (%) was surprisingly high (mean 41% ± 34; median 33.4%) and significantly different ( P < 0.001) from the corresponding value in healthy human subjects (mean 0.98% ± 0.6; median 1.3%). 2. 2. It was negatively correlated with 18:2n-6 (r = −0.76, P < 0.001), 18:2n-6 20:4n-6 (r = −0.70, P < 0.005) and 18:2n-6 22:6n-3 (r = −0.44, P < 0.025) in the ethanolamine phosphoglycerides of the erythrocytes. 3. 3. A shift in the n-6 n-3 balance in favour of the n-3 in the fatty acid composition of the membrane may have predisposed the red blood cells to haemolysis. 4. 4. An increase in 22:6 n-3, and a decrease of 20:4 n-6 and 18:2 n-6, may have been the result of feeding a diet containing fish products rich in long chain n-3 fatty acids.

TISSUE RESPONSES TO HYPEROXIA: Biochemistry and Pathology

An animal model was established to study the toxic effects of hyperoxia and the consequent changes in intracellular antioxidant status. Superoxide dismutase, catalase and glutathione peroxidase activities were measured in erythrocytes, liver and lung, in addition to cellular glutathione concentrations and its associated metabolism. Overt cellular damage was assessed biochemically by measurement of lipid peroxidation, hydrogen peroxide-induced haemolysis and osmotic fragility. Pathological changes were assessed by light and electron microscopy. Up to 11 days exposure of rats to 80% oxygen was not lethal, but resulted in overt cellular damage to red blood cells (haemoglobin concentration decreased from 13.8 +/- 1.4 (SD) g dl-1 to 12.4 +/- 0.5 g dl-1; hydrogen peroxide-induced haemolysis increased from 7.7 +/- 1.6% to 75.1 +/- 13.5% after 11 days of hyperoxia) and to cells of lung (4-fold increase in lipid peroxidation) as well as a biochemical adaptation to the increased concentration of oxygen metabolites (superoxide dismutase increased 3-fold, catalase 5-fold and glutathione peroxidase 2-fold). It is suggested that red cell hydrogen peroxide-induced haemolysis and reduced glutathione concentration may be useful indicators of oxidant stress in the clinical situation.

Antiplatelet effect of hsien-ho-t'sao (Agrimonia pilosa).

The water extract of Hsien-Ho-T'sao (HHT) produced a dose-dependent inhibition on collagen-induced aggregation of platelet-rich plasma (PRP). The IC50 was about 3.5 mg/ml. In addition, HHT inhibited also the aggregation induced by ADP, A23187 or arachidonate in PRP. Greater inhibition was observed in the preparation of washed platelets. Increase of the calcium concentration in medium could not overcome the inhibitory effect of HHT. ATP release from platelets induced by collagen or A23187 was inhibited by HHT. In the presence of EDTA, ATP release caused by thrombin or A23187 was also inhibited by HHT. Malondialdehyde and thromboxane B2 formation was greatly inhibited by HHT in platelets challenged by collagen and thrombin. In arachidonate-stimulated platelets, thromboxane B2, but not malondialdehyde formation was inhibited. HHT showed more marked inhibition on aggregation in the presence of indomethacin, creatine phosphate/creatine phosphokinase or a combination of both. Hydrogen peroxide-induced hemolysis was marked reduced by HHT. It was concluded that HHT might have some membrane-active properties which interfered with the activation of phospholipase A2.

Inhibitors of hydrogen peroxide-induced haemolysis of bovine erythrocytes

The optimal conditions for the haemolysis of bovine erythrocytes by H 2O 2 have been established. The parameters were concentration of erythrocytes, H 2O 2 concentration, time, and influence of the solvent in which the substances tested were dissolved. Some inhibitors of this oxidative haemolysis have been employed to serve as model substances for further antihaemolytic investigations with natural products.

Effect of Monolauryl Succinate, Lauryl Alcohol and Succinic Acid on Hemolysis and Liver Mitochondrial Lysis in Vitro in Chicks

In order to elucidate the mechanism of dilauryl succinate-induced vitamin E deficiency in vivo in chicks, I conducted experiments to study the effect of the metabolites of dilauryl succinate in chicks on hemolysis and the liver mitochondrial lysis, while neither lauryl alcohol nor succinic acid induced mitochondrial lysis in vitro in chicks. These were monolauryl succinate, lauryl alcohol and succinic acid. Monolauryl succinate induced both hemolysis and mitochondrial lysis at the same concentration as that of monolauryl succinate. At the concentration of 40 times that of monolauryl succinate, lauryl alcohol induced hemolysis but succinic acid did not. Neither alpha-tocopherol, selenium nor combination of both exerted preventive effects on the reactions of monolauryl succinate or lauryl alcohol. Monolauryl succinate-induced mitochondrial lysis was not accompanied by lipid peroxidation. The observations were consistent with those in Tween 20-induced hemolysis and mitochondrial lysis, but not with those in hydrogen peroxide-induced hemolysis or with those in glutathione-induced mitochondrial lysis. It was suggested that dilauryl succinate-induced vitamin E deficiencies in vivo in chicks are mainly due to the surface activity, which is not relevant to peroxide formation, of monolauryl succinate and partly to that of lauryl alcohol.

Increased Erythrocyte Fragility with Hydrogen Peroxide in Vitamin E-Deficient Chickens

INTRODUCTIONIN THE course of studying vitamin E deficiency induced in chicks it became apparent that the lack of detailed information about the pathogenesis of the disorder inhibits the development of precise methods of treatment and control of the condition, impairs the understanding of the metabolic role of vitamin E, and precludes the utilization of the disease in chicks as a convenient experimental model for elucidating certain pathobiologic phenomena.An increased susceptibility to lysis in non-physiologic solutions of red blood cells from mammals with subclinical vitamin E deficiency has been well substantiated in reports by György (1949), and Horwitt et al. (1956), and others; however, the in vitro lytic susceptibility of erythrocytes from vitamin E-deficient chicks has been incompletely studied (Christensen et al., 1955). Since substantial differences exist among various vertebrate species with regard to other manifestations of avitaminosis E, a study was made of hydrogen peroxide-induced hemolysis of red cells.