RESUMEN
Blood and its components are stored to meet the demands of blood transfusion. Erythrocytes undergo progressive modifications during storage known as storage lesions. Storage solutions were developed to improve shelf life and extend red cell viability. Therefore, the objective of this study is to analyze the effects of AS-7 on young and old erythrocytes during storage. Blood was collected from the blood bank at Kempegowda Institute of Medical Sciences (KIMS) hospital, Bengaluru. Erythrocytes were isolated from whole blood and separated based on its age using Percoll density gradient. The young and old erythrocytes were stored in AS-7 for 35 days and every 5th day, oxidative stress markers - Hemoglobin (Hb), Oxidative Hemolysis, Mechanical Fragility, Sialic Acid, Superoxides, Glucose, Lactate Dehydrogenase (LDH), Glutathione, antioxidant capacity (TACCUPRAC), Plasma Membrane Redox System (PMRS), antioxidant enzymes, lipid peroxidation, and protein oxidation products were assessed. Hb, glucose, TACCUPRAC, and superoxide dismutase reduced, while oxidative hemolysis, mechanical fragility, protein oxidation, and lipid peroxidation products increased in young and old cells over storage. LDH, PMRS, catalase, advanced oxidation protein products, and conjugate dienes were significant in old cells from day 5 itself, whereas in young cells towards the end of storage (from day 25). Oxidative insult was higher in old cells compared to young cells. AS-7 was beneficial to young erythrocytes during storage and thus laying the foundation for the possibilities of utilizing young cells as models for storage studies.
Asunto(s)
Conservación de la Sangre/métodos , Eritrocitos/metabolismo , Ácido N-Acetilneuramínico/metabolismo , Estrés Oxidativo/efectos de los fármacos , Eritrocitos/citología , HumanosRESUMEN
BACKGROUND: During storage of erythrocytes, storage lesions are formed that reduce the safety and efficacy of the stored blood. Thus, there is a need to understand the changes that occur during storage. Most studies have focused on storage of a mixed population of erythrocytes. The aim of this study is to analyze the changes in young and old erythrocytes over the course if storage. MATERIALS AND METHODS: Blood was collected from the blood bank at the Kempegowda Institute of Medical Sciences (KIMS) Hospital (Bengaluru, India) and stored for 35 days in CPDA-1 at 4°C. Every 5 days, erythrocytes were separated based on the blood's age using a Percoll-BSA gradient. Young and old erythrocytes obtained were used for analysis of the following oxidative stress (OS) markers: hemoglobin (Hb), hemolysis, mechanical fragility, antioxidant enzymes (superoxide dismutase and catalase [CAT]), superoxides, sialic acid, glutamic oxaloacetate transaminase (GOT), glucose, plasma membrane redox system (PMRS), total antioxidant capacity-cupric ion reducing antioxidant capacity assay (TACCUPRAC), lactate dehydrogenase (LDH), lipid peroxidation products (malondialdehyde [MDA] and conjugate dienes), and protein oxidation products (advanced oxidation protein products and protein sulfhydryls). RESULT: Young cells had higher amounts of Hb, sialic acid, GOT, LDH, TACCUPRAC, CAT, and superoxides compared to old cells. Old cells, however, had higher PMRS and MDA levels with respect to young cells. DISCUSSION: Young cells could endure OS during storage more efficiently than old cells. In conclusion, the efficacy of stored blood depends on the ratio of young cells in the population. This study highlights the prospects of storing young erythrocytes for a prolonged shelf life of blood.
RESUMEN
The effects of rejuvenation on the subpopulation of stored erythrocytes have not been explored. This study aims at determining the influence of rejuvenation on young and old erythrocytes of stored blood. Prior studies have shown the disappearance of young cells after day 20 of storage. Blood was stored in CPDA-1 for 35 days and erythrocytes were isolated on 25th, 30th and 35th day, revitalized using rejuvenation solution (PIPA), and separated into young and old erythrocytes using Percoll-BSA density gradient. Erythrocyte, oxidative stress and antioxidant capacity markers were assessed in the hemolysate. Young erythrocytes could be isolated beyond day 20 of storage, after rejuvenation. Antioxidant capacity of both youngRej (rejuvenated young cells) and oldRej (rejuvenated old cells) increased while superoxides decreased resulting in lower levels of protein oxidation & lipid peroxidation. Rejuvenation reduced storage lesion and maintained membrane sulfhydryls in both young and old erythrocytes, however, it could not restore sialic acids. Rejuvenation had more impact on youngRej. A higher young: old cell ratio would be beneficial for transfusion. This study gives a comparative analysis of rejuvenation on erythrocyte aging in banked blood, thus opening new avenues towards better blood bank practices.
RESUMEN
OBJECTIVE: Plasma acts as a good indicator of oxidative stress in blood. L-Carnitine is an antioxidant that reduces metabolic stress in cells, thereby providing a protective effect against oxidative stress (OS). L-Carnitine as an additive in storage has not been explored. Thus, this study attempts to analyze the role of L-carnitine in blood storage solution, citrate phosphate dextrose adenine (CPDA)-1, through OS markers including antioxidant enzymes, lipid peroxidation, and protein oxidation. MATERIALS AND METHODS: Blood was collected from male Wistar rats and stored in CPDA-1 solution with L-carnitine (10 mM, 30 mM, and 60 mM: groups LC 10, LC 30, and LC 60, respectively) and without L-carnitine (control group). Plasma was isolated every 5th day and the OS markers were analyzed. RESULTS: Superoxide dismutase (SOD) and sulfhydryl (SH) increased over storage in controls, LC 30, and LC 60. Catalase increased in LC 30 and LC 60 during storage. Thiobarbituric acid reactive substances (TBARS) and protein carbonyl (PrC) levels in all groups increased initially and reduced towards the end of storage. SOD and SH levels were maintained while TBARS and PrC levels increased in LC 10. CONCLUSION: L-Carnitine was beneficial in terms of increased antioxidant capacity and SH and decreased lipid peroxidation. This forms the basis for further studies on L-carnitine as a constituent in storage solutions.
Asunto(s)
Antioxidantes/metabolismo , Conservación de la Sangre/métodos , Carnitina/metabolismo , Plasma/metabolismo , Adenina/metabolismo , Animales , Catalasa/metabolismo , Citratos/metabolismo , Glucosa/metabolismo , Peroxidación de Lípido , Masculino , Estrés Oxidativo , Fosfatos/metabolismo , Plasma/enzimología , Ratas Wistar , Superóxido Dismutasa/metabolismo , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
BACKGROUND/AIM: Curcumin, a naturally occurring antioxidant, shows a wide variety of medicinal properties. The possibility of utilizing curcumin as an additive in storage solutions of blood has not been explored. The purpose of this study was to analyze the effect of curcumin on erythrocytes during storage. MATERIALS AND METHODS: Blood obtained from rats was stored (4 °C) for 20 days in citrate-phosphate-dextrose-adenine-1 solution. Samples were divided into four groups: 1) Controls; 2) Curcumin 10 mM; 3) Curcumin 30 mM; and 4) Curcumin 60 mM. Every fifth day, hemoglobin, superoxide, antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase (GSH-Px)), lipid peroxidation (conjugate dienes and malondialdehyde (MDA)), protein oxidation (advanced oxidation protein products (AOPP) and sulfhydryls (P-SH)), and hemolysis were analyzed. RESULTS: Hemoglobin was successfully maintained, while superoxide dismutase increased initially and decreased towards the end of storage. Superoxide, catalase, GSH-Px, conjugate dienes, and AOPP were lower in the curcumin groups than they were in the controls. MDA was higher in the curcumin groups than in the controls. P-SH increased in the curcumin groups, while hemolysis increased in all groups. CONCLUSION: Curcumin maintained hemoglobin and modulated antioxidant enzymes throughout storage. However, curcumin could not protect all proteins and lipids from oxidative damage completely. This study opens up new avenues for using curcumin, in combination with other antioxidants, as a component in storage solutions.