Objective: To investigate the changes of CD71(+) nucleated erythrocyte apoptosis, cytochrome C (Cyt-C) and mitochondrial membrane potential (MMP) in bone marrow of chronic mountain sickness (CMS). Methods: 14 patients with CMS and 15 patients with simple old fracture were divided into CMS group and control group, respectively.Bone marrow mononuclear cells (BMMNC) were separated, 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, and Cyt-C mRNA was detected by RT-qPCR, MMP was detected by JC-1 staining flow cytometry. Results: The apoptotic index of CD71(+) nucleated erythrocytes was (1.9±1.4)% in the CMS group, and was (3.2±1.5)% in the control group, with significant difference between the two groups (P<0.05). 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<0.01). The MMP was (5.0±2.2) in the CMS group, and was (3.3±0.9) in the control group, with significant difference between the two groups (P<0.05). The apoptotic index of CD71(+) nucleated erythrocytes was negatively correlated with hemoglobin in CMS group (r=-0.569, P=0.034). But there was no significant correlation among apoptosis index, MMP and Cyt-C mRNA. Conclusions: The apoptosis index of CD71(+) nucleated erythrocytes decreased in CMS patients, which was negatively correlated with the level of 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 MMP increased and Cyt-C mRNA expression decreased in CD71(+) nucleated erythrocytes of CMS patients, which suggests that the change of mitochondrial pathway of apoptosis might be involved in the down-regulation of CD71(+) nucleated erythrocytes apoptosis in CMS patients.But there was no significant correlation among CD71(+) nucleated erythrocyte apoptosis index, MMP and Cyt-C mRNA levels, which indicates that the mechanism of CD71(+) nucleated erythrocytes apoptosis is complex in CMS.
目的： 探讨慢性高原病(CMS)患者骨髓CD71(＋)有核红细胞凋亡和细胞色素C(Cyt－C)及线粒体膜电位(MMP)的变化情况。 方法： 以14例CMS患者及15例单纯陈旧性骨折患者分别为CMS组和对照组，提取骨髓单个核细胞(BMMNC)，CD71单克隆抗体标记、Annexin V－FITC/PI染色，流式细胞术检测CD71(＋)有核红细胞的凋亡指数，磁珠分选法分选CD71(＋)有核红细胞并以RT－qPCR检测Cyt－C mRNA的表达水平；JC－1染色流式细胞术检测CD71(＋)有核红细胞MMP。 结果： CMS组骨髓CD71(＋)有核红细胞凋亡指数为(1.9±1.4)%明显低于对照组(3.2±1.5)%，差异有统计学意义(P<0.05)。CMS组Cyt－C mRNA表达水平(0.72±0.14)明显低于对照组(1.00±0.15)，差异有统计学意义(P<0.01)。CMS组MMP(5.0±2.2)明显高于对照组(3.3±0.9)，差异有统计学意义(P<0.05)。CMS患者骨髓CD71(＋)有核红细胞凋亡指数与血红蛋白水平呈负相关(r＝－0.569，P＝0.034)，凋亡指数、MMP及Cyt－C mRNA间未发现明显相关性。 结论： CMS患者骨髓CD71(＋)有核红细胞凋亡指数下降，且与血红蛋白水平呈负相关，说明有核红细胞凋亡下调可能与CMS红细胞积累有关；CD71(＋)有核红细胞MMP增高，Cyt－C mRNA相对表达水平明显降低，提示线粒体通路变化参与CMS患者骨髓CD71(＋)有核红细胞凋亡下降机制，而CD71(＋)有核红细胞凋亡指数、MMP、Cyt－C mRNA水平之间没有发现明显的相关性，提示CMS造血细胞凋亡机制的复杂性。.
Keywords: Apoptosis; CD71(+) nucleated erythrocyte; Chronic mountain sickness; Cytochrome C; Mitochondrial membrane potential.