The Study On Eryptosis Leading To Anemia In Patients With SLE

Objective : To study eryptosis in systemic lupus erythematosus(SLE)patient with anemia and further explore possible mechanisms.Methods : This experiment was devied into erythrocyte experiment and plasma incubation experiment.We select SLE patients as experimental group and healthy blood donors as control group.In erythrocyte experiment,20 fresh blood samples were draw from every group.We detected hematological parameters of these blood sample,separated erythrocytes and made up suspended erythrocytes to study eryptosis.After cells were stained by annexinV-fluorescein isothiocyanate(FITC),the apoptosis rate and volume size were assessed using flow cytometry.Meanwhile,observe the shape of apoptosic cells by confocal laser scanning microsco(CLSM)。Fluorescent probe Fluo-3/AM and DCFH-DA were respectively used to quantificated intracellular calcium and reactive oxygen species(ROS).Ceramide abundance was measured using species-specific secondary antibodies labeled with fluorescent.The two groups were compared in terms of above biochemical indicators.The influence of plasma of SLE patients on eryptosis by plasma incubation experiment.We gather 20 fresh blood samples were draw from every group and separated plasma to incubate erythrocytes(O+ blood group)from healthy individuals.Using the same methods,measured and compared above indicators after 24 hours.In addition,measured and compared plasma total antioxidant capacity between two groups by ELIASA.Results : In erythrocyte experiment,the SLE patients revealed a hemoglobin concentration and hematocrit,which were significantly lower than that in healthy volunteers(***p<0.001,p<0.001 respectively),the percentage of eryptosis was significantly higher in SLE patients than in the healthy volunteers(***p<0.001),which was paralleled by an increase in cytosolic Ca2+ levels and oxidative stress.The cell volume was significantly higher(**p<0.01)in erythrocytes from healthy volunteers than from the SLE patients.We can observe fluorescence signal in apoptotic cell by CLSM.In incubation experiment,the cytosolic Ca2+ levels were significantly higher(*p<0.05)in experimental group than in control group.However,ceramide,ROS abundance,cell volume and the plasma total antioxidant capacity have no significant differences between the two group.Conclusions : Eryptosis is enhanced in SLE and may contribute to anemia.The probable underlying mechanisms may be an excessive formation of ROS in erythrocytes.Also,some plasma components may trigger eryptosis by increasing the cytosolic Ca2+ concentration.