EFFECTS OF REDUCED GLUTATHIONE ON THE OXYGENATED AND PRESERVED RED CELLS

ITS ANTI-HEMOLITIC AND ANTI-OSMOTIC FRAGILITY EFFECTS

Abstract

Denaturation of hemoglobin is one of the intracellular factors which disturb red cell metabolism and lead to the hemolysis. Although there are many pathways to reduce methemoglobin to hemoglobin in packed cell, it seems to be most important to keep hemoglobin intact during storage.
Oxydation of hemoglobin to methemoglobin by the oxygen has been shown to occur in reduced hemoglobin rather than the oxygenated hemoglobin by Brooks in 1948.
Sase reported in 1965 that oxygenated hemoglobin decreased methemoglobin products as compared with half to half mixture of oxygenated and reduced hemoglobin.
The red cell membrane is known to possess many sulfhydryl units, especially in the inner layer.
Inhibition of the sulfhydryl units of the red blood cell has been shown to result hemolysis. (H. S. Jacob & J. H. Jandl)
Reduced glutathione is also effective in maintaining sulfhydryl groups of the red blood cell membrane.
In the present study, the influence of the oxygenation and an addition of reduced glutathione on the red cell fragility and hemolysis was determined.
The experiments consisted of the following four groups.
1) Venous blood collected into ACD solution (VA)
2) Venous blood collected into ACD solution with reduced glutathione (VAGSH)
3) Oxygenated blood in ACD solution (OA)
4) Oxygenated blood in ACD solution with reduced glutathione (OAGSH)
Results
1) Plasma free hemoglobin during a five week storage
It was least in the OA and OAGSH groups.
2) Red cell fragility test
This was tested against a series of 0.9% to 0.2% sodium chloride solution. At the level of 0.8% NaCl, an addition of GSH was effective to keep weak resistant red cell from destruction.
OAGSH tolerated well at the level of 0.8%, however, damage of the cell membrane of the well resistant young red cell was prominent.
3) Serum lactate and pyruvate levels
On lactate to pyruvate ratio, except control group (VA), the other stored blood groups reached to its maximum value within 2 weeks and reduced gradually to its initial level.
On the contrary, VA reached to its maximum value after 3 weeks storage. The reduced glutathione increased lactate to pyruvate ratio higher but did not change the time to its maximum value.
Conclusion
Oxygenation and the addition of reduced gluthatione have proven to give a veneficial effects on anti-hemolysis and fragilty change during a 5 weeks RBC storage.