| Objective Chronic mountain sickness(CMS)is a common disease in high altitude area,which is a serious threat to human health.However,due to the complexity of its pathogenesis,there is no definite conclusion on the mechanism of occurrence and development of CMS up to now.The research on the prevention and treatment of CMS is still in the exploration stage.The accumulation of red blood cells is one of the main features of the CMS,but the effect of nucleated red blood cells apoptosis in which is indeterminate.Therefore,the aims of this paper were to investigate the changes of apoptosis,Cytochrome C(Cyt-C)and mitochondrial membrane potential(MMP)in CD71~+nucleated erythrocytes in patients with CMS,and to further explore the possible role of apoptosis in the path of mitochondrion of CD71~+nucleated erythrocytes in CMS patients,and to provide more scientific basis for the pathogenesis and clinical intervention of CMS.Methods The objects were outpatients and inpatients of Qinghai University Affiliated Hospital from 2015 to 2017.The 14 CMS patients were CMS group,with an average age of(42.5±6.2)years,which were diagnosed by“Consensus statement on chronic and subacute high altitudediseases”in“VI World Congress on Mountain Medicine and High Altitude Physiology”(2004,Qinghai,China);The 15 patients with simple old fracture were control group,with(41.5±11.4)years old of average age.There was no significant difference in the age between the two groups(P>0.05).The acute and chronic infections,cancer,immune-related diseases,and primary diseases of heart,lung,liver and kidney were excluded.Bone marrow was collected and the bone marrow mononuclear cells(BMMNCs)were separated.The BMMNCs were marked with CD71 monoclonal antibody and stained with Annexin V-FITC/PI.Then the apoptotic index of CD71~+nucleated erythrocytes was determined by flow cytometry.CD71~+nucleated erythrocytes were sorted out by magnetic column separation technique.Cyt-C mRNA and MMP were detected by RT-qPCR and flow cytometry with JC-1staining,seperately.Results The results in the present study showed that the apoptotic index of CD71~+nucleated erythrocytes were(1.92±1.41)%in the CMS group and(3.16±1.49)%in the control group,with significant difference between the two groups(P=0.031).The expression of Cyt-C mRNA was(0.72±0.14)in the CMS group,and was(1.00±0.15)in the control group,with significant difference between the two groups(P<0.001).The MMP were(4.96±2.19)and(3.29±0.93)in the CMS group and the control group,separately,there was significant difference between the two groups(P=0.017).The apoptotic index of CD71~+nucleated erythrocytes was negatively correlated with hemoglobinin CMS group(r=-0.569,P=0.034).But there was no significant correlation among apoptosis index,MMP and Cyt-C mRNA.Conclusion The apoptosis index of CD71~+nucleated erythrocytes decreased in CMSpatients,whichwasnegativelycorrelatedwiththelevelof hemoglobin,indicating that the decline of apoptosis index of CD71~+nucleated erythrocytes may be related to the accumulation of red blood cells in CMS.The increased MMP and decreased Cyt-C mRNA suggest that the change of mitochondrial pathway of apoptosis might be involved in the down-regulation of apoptosis in CD71~+nucleated erythrocytes of CMS patients.The result of no any significant correlation among apoptosis index,MMP and Cyt-C mRNA levels in CD71~+nucleated erythrocytes indicates that the mechanism of CD71~+nucleated erythrocytes apoptosis is complex in CMS. |
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