Hydrogen Rich Saline Effect On Immune Mediated Aplastic Anemia Model Mice

ObjectiveThe paper is to study the function of hydrogen rich saline combined with cyclosporineon immune mediated aplastic anemia model mice bone marrow hematopoietic function andcytokines, and to investigate the hydrogen-rich saline on aplastic anemia and protectiveeffect of anti-inflammatory and antioxidant mechanism, so as to provide the basis foreconomic and effective method for the treatment of aplastic anemia.MethodsEstablishing aplastic anemia mice model according to the method of Yao Jun.BALB/c mice shall be treated with whole body irradiation with X rays (total dose5.0Gy)(absorbed dose rate is1.21Gy/min, irradiation time is248s), then within4h injectingDBA/2mice through caudal vein with lymphocyte suspension0.2ml/mouse. Grouping oftesting animals: normal group, control group (group AA), H2group (Hydrogen rich saline10ml/kg), CsA group (oral CsA), CH group (CsA+Hydrogen rich saline10ml/kg),14~28days after model setting, according to the group type taking medicine injection. Adoptingthe hematology analyzer for measurement of mice on the first,7th,14th,21st and28th day.On the28th day, taking plasma of mice for measurement of SOD, MDA, IL-3, IL-6,TNF-α and TGF-β. Fixing the femur and spleen of mice in4%paraformaldehyde, paraffinHE staining protein expression in spleen IFN-γ immunohistochemistry mice.Results1.The activity of mice in H2group is slow after model making, intake of food andwater is poor, the survival rate of mice is50%. While the activity of mice in hydrogen richsaline combined with cyclosporine groupis basically normal with good food intake, thesurvival rate of mice is80%, the weight of mice grows evidently. Through the observationof the indicators such as general physical condition, mental, appetite, weight gain,increased survival observed indicators, it is obvious that the hydrogen rich saline combined with cyclosporine is better than cyclosporine or hydrogen rich saline for treatment.2.The blood indices such as leukocyte, hemoglobin and platelet increased in thehydrogen rich saline combined with cyclosporine group, bone marrow pathological HEstaining cells increased a lot, hematopoietic function improved significantly (P<0.05)higher than other experimental groups.3.The plasma levels of SOD of mice showed that the SOD levels in hydrogen-richsaline combined with cyclosporine in the experimental group is significantly (P<0.05)higher than other experimental groups. The MDA levels of mice in hydrogen-richcombined with cyclosporine group is significantly (P<0.05) lower than other experimentalgroups.4.ELISA method for measurement of plasma IL-3, TNF-α and TGF-β levels, thehematopoietic positive regulation of cytokine IL-3levels of mice in the CH group weresignificantly (P<0.05) higher than other experimental groups. The negative regulation ofinflammatory factor TNF-α and TGF-β levels of mice in the CH group were significantly(P<0.05) lower than other experimental groups.5.As for the immunohistochemical spleen IFN-γ compared with the otherexperimental groups, the level of IFN-γ positive cells in the spleen rate in the CH groupreduced with significant difference (P<0.05).Conclusion1.Hydrogen rich saline can promote the recovery of aplastic anemia in peripheralblood cells and bone marrow.2.Hydrogen rich saline aplastic anemia can decrease the production of inflammatorycytokines in aplastic anemia, promote positive hematopoietic factor IL-3generation,inhibite negatively regulates cytokine IL-6, TNF-α and TGF-β levels. The level of IFN-γpositive cells in the spleen rate is reduced. It has anti-inflammatory effect.3.Hydrogen rich saline aplastic anemia can inhibit the generation of oxygen freeradicals, reduce SOD consumption and reduce lipid peroxidation product MDA generation,it has antioxidant effects.