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Článek online

DMT1-mutant erythrocytes have shortened life span, accelerated glycolysis and increased oxidative stress

Cellular physiology and biochemistry. 2014 ; 34 (6) : 2221-31. [pub] 20141204

Cell Physiol Biochem
ISSN 1421-9778
Medvik
Zdroj

BACKGROUND/AIMS: Deficiency of the divalent metal transporter 1 (DMT1) leads to hypochromic microcytic anemia. We have previously shown that DMT1 deficiency impairs erythroid differentiation and induces apoptosis of erythroid cells. Here we analyzed metabolic processes and survival of mature erythrocytes in order to address potential involvement of erythrocyte defect in the pathophysiology of the disease. METHODS: FACS analysis was used to determine the half-life of erythrocytes (CFSE fluorescence), phosphatidylserine exposure (Annexin V binding), cytosolic Ca(2+) (Fluo3/AM fluorescence) and reactive oxygen species (ROS; DCF fluorescence). Enzyme activities were determined by standard biochemical methods. The concentration of ATP and ADP was measured on HPLC-MS/MS. RESULTS: We observed an accelerated clearance of CFSE-labeled DMT1-mutant erythrocytes from circulating blood when compared to wild-type erythrocytes. In vitro, DMT1-mutant erythrocytes showed significantly increased Annexin V binding after exposure to hyperosmotic shock and glucose depletion. Despite exaggerated anti-oxidative defense, higher ROS levels were present in DMT1-mutant erythrocytes. Accelerated anaerobic glycolysis and reduced ATP/ADP ratio detected in DMT1-mutant erythrocytes indicate enhanced demand for ATP. CONCLUSIONS: We propose that DMT1 deficiency negatively affects metabolism and life span of mature erythrocytes; two other aspects of defective erythropoiesis which contribute to the pathophysiology of the disease.

MeSH
adenosindifosfát krev MeSH
adenosintrifosfát krev MeSH
apoptóza genetika MeSH
erytrocyty metabolismus patologie MeSH
erytropoéza MeSH
glykolýza MeSH
hypochromní anemie krev genetika patologie MeSH
lidé MeSH
modely nemocí na zvířatech MeSH
mutace MeSH
myši MeSH
oxidační stres * MeSH
proteiny přenášející kationty krev nedostatek genetika MeSH
reaktivní formy kyslíku krev MeSH
zvířata MeSH
Check Tag
lidé MeSH
myši MeSH
zvířata MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH

Článek

In vitro parameters of cryopreserved leucodepleted and non-leucodepleted red blood cells collected by apheresis or from whole blood and stored in AS-3 for 21 days after thawing

Blood purification. 2014 ; 12 Suppl 1 (-) : s199-203.

Blood Purif
ISSN 1723-2007
Medvik
Zdroj

BACKGROUND: The aim of the study was to evaluate the in vitro quality of cryopreserved red blood cells obtained from different sources with or without leucodepletion and stored at 4±2 °C in AS-3 for up to 21 days. MATERIALS AND METHODS: Red blood cells were collected by four methods: double erythrocytapheresis, whole blood collection with buffy coat removal, double erythrocytapheresis with in-line leucofiltration, or whole blood collection with in-line leucofiltration. All four types of red blood cells were frozen in 40% glycerol after collection and stored at a temperature below -65 °C for at least 30 days, thawed, deglycerolised and subsequently reconstituted in AS-3. The in vitro haematological and biochemical properties of the thawed red blood cells were tested on days 0, 7, 14, and 21 after deglycerolisation and reconstitution. RESULTS: Overall, 72 units were processed. Leucodepletion of cryopreserved red blood cells units reduced haemolysis, lowered ammonia concentration, preserved pH and osmolality and led to sustained higher concentrations of ATP. In contrast, the source of red blood cells (apheresis or whole blood) did not affect their quality. DISCUSSION: The quality of all investigated red blood cells units was the same as or even better than that of erythrocytes obtained from double erythrocytapheresis with a 24-hour survival of at least 86% after up to 3 weeks of storage in AS-3.