Ozone easily reacts with unsaturated fatty acids in the phospholipid bilayer membrane of red blood cells, accompanying the breakage of unsaturated fatty acid chains and producing short chain hydroperoxides, thereby affecting cellular glycolysis. After ozone administration, glycolysis and ATP content in red blood cells increased. The increase of ATP induced by ozone in red blood cells can stabilize cell membrane tension and enhance machine resistance, while the trace amount of ATP outside the cell can generate a series of rapid ozone reactions, such as vasodilation and energy storage during inflammation. Through in vitro and in vivo studies, it has been proven that ozone can directly interfere with the metabolism of red blood cells. The mechanism is through a specific reaction with the double bond of unsaturated fatty acids, leading to the breakdown of phospholipids in the red blood cell membrane and the entry of substances such as hydrogen peroxide into the cell. Red blood cells immediately respond, initiating the peroxidation detoxification mechanism as an oxidative shield system, ultimately protecting the divalent iron in hemoglobin from oxidation. At the same time, ozone can maintain the balance of hemoglobin/oxygen and activate red blood cell metabolism by affecting the glutathione system. In the presence of glutathione reductase and glutathione peroxidase, peroxide analysis is carried out. After ozone therapy, the content of reduced glutathione immediately decreased and returned to its previous concentration after about 15 minutes. Scientific research has shown that the connection and regeneration of the glutathione system are related to the increase of glucose-6-phosphate dehydrogenase and the activation of the pentose phosphate pathway, and the activation of the pentose phosphate pathway ultimately activates all red blood cell metabolism.
Ozone regulates the efficacy of red blood cells and increases oxygen release by affecting 2,3-DPG. 2,3-triphosphate glycerol has a special effect on red blood cells. As an oxygen transporter, it must store the oxygen it carries and release it at necessary locations for utilization. However, the affinity of hemoglobin for oxygen largely depends on 2,3-DPG. In healthy individuals, 2,3-DPG appears in the same molar amount as hemoglobin and can enter the center of its tetrahedral layout while releasing four molecules of oxygen. After ozone therapy, the content of 2,3-DPG increased, which reduced the affinity of red blood cells for oxygen. This means that red blood cells are more likely to release oxygen, increasing peripheral oxygen supply. Other ozone can cause morphological changes in red blood cells, improve their flexibility, and enhance their blood rheological properties.

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