Nutritional Rehabilitation Of Mitochondrial Aberrations In Aplastic Anaemia

Objective: BALB/c AA mice were induced by a combination of hypodermic injections of acetylphenylhydrazine, X-rays and intraperitoneal injections of cyclophosphamide. We explored the nutritional rehabilitation of mitochondrial aberrations in aplastic anaemia.Methods:After one week adaptation period, 100 BALB/c mice were randomly divided into five groups with twenty in each group: the control group; the AA group; the three different doses of the nutrient-mixture-supplemented diet groups. AA mice were induced by a combination of hypodermic injections of acetylphenylhydrazine (100 mg/kg), X-rays (2.0 Gy) and intraperitoneal injections of cyclophosphamide (80 mg/kg). We treated these mice with nutrient mixture-supplemented diets in a dose-dependent manner (1445.55, 963.70, 674.59 mg/kg per d) by intragastric administration. The control and AA model groups were given a physiological saline (10 ml/kg per d)-supplemented diet by intragastric administration. After 7 weeks, the mice were killed by cervical dislocation and then were determined in the following items.(1) Analysis of peripheral blood (RBC,WBC,plt,Hb); (2) Transmission electron microscopy (TEM) studies; (3) Mitochondrial membrane potential (MMP) assay; (4) Analysis of cytochrome c protein levels; (5) mtDNA content assay.Results:(1) A significant peripheral blood pancytopenia was evident in AA mice (*p<0.05). The peripheral blood parameters in the three different doses of the nutrient mixture groups were higher than in the AA group in a dose-dependent manner (#p<0.05).(2) The size and shape of mitochondria of the control groups were normal. Mitochondria had enlarged forming globular structures accompanied by disruption or disappearance of the cristae in the AA group. Remarkably, mitochondrial ultrastructure was restored in the nutrient mixture groups, comparing with AA groups.(3) The results showed that the Rh123 fluorescence of bone marrow cells, splenocytes and hepatocytes in the AA group were lower than in the control group (p < 0.05 ). However, the Rh123 fluorescence of three kinds of cells in the three different doses of the nutrient mixture groups were higher than in the AA group on dose-dependent ( p < 0.05 ).(4) Cytochrome c expression was variable among the mice models, with higher cytochrome c level detected in AA group cells and no detectable level in control group cells. However, the cytochrome c levels of three kinds of cells in the nutrient mixture groups were markedly lower than in AA group on dose-dependent ( p < 0.05 ).(5) The mtDNA content of bone marrow cells, splenocytes and hepatocytes in the AA group was lower than in the control group (p < 0.05 ). However, the mtDNA content of three kinds of cells in the three different doses of the nutrient mixture groups were higher than in the AA group in dose-dependent ( p < 0.05 ).Conclusion:(1) The nutrient mixture had an effect on peripheral blood cell levels in AA mice.(2) The nutrient mixture may promote the rehabilitation of mitochondrial aberrations in AA mice.