| Objective: The excessive accumulation of red blood cells is one of the main characteristics of CMS,Chronic mountain sickness(CMS)is a common disease in plateau area.but its mechanism has not been fully elucidated.Apoptosis plays an important role in hematopoietic regulation in physiological conditions.Our previous studies have found that hematopoietic cell apoptosis was downregulated in CMS.This study was focus on erythroblasts apoptosis in bone marrow of CMS,and the expression of Bcl-2,Bcl-xl and Bax m RNA.The aim of the study was to explore the change of Bcl-2 family members in mitochondrial signaling pathway and the effect on the apoptosis of erythroblasts in CMS.Method: 26 male patients with(50.2±11.4)years old of average age(CMS group)were diagnosed by “Consensus statement on chronic and subacute high altitude diseases” in “VI World Congress on Mountain Medicine and High Altitude Physiology”(2004,Qinghai,China).12 male healthy adults served as the control group.They were Chinese Hans with(48.5±12.6)years old of average age.The diseases of heart,lung,liver and kidney were excluded in all subjects.Their bone marrow were collected and anticoagulated by heparin.Bone marrow mononuclear cells(BMMNCs)were isolated by lymphocyte separation and counted.The apoptotic rate of CD71+ cells was determined using flow cytometry with signed by CD71 monoclonal antibody and Annexin V-FITC/PI.CD71+ cells in BMMNCs was sorted by immunomagnetic bead sorting and then the RNA was extracted.The expressions of Bcl-2,Bcl-xl and Bax mRNA were detected by real-time PCR.Results: 1.The expression of Bax m RNA in CMS group was lower than that in control group(0.7067±0.01154 vs 1.0006±0.05063).There was significant difference between the two groups(t=13.866,p<0.001);2.The expression of Bcl-xl m RNA in CMS group was lower seemingly than that in control group(0.8581±0.27270 vs 1.0560±0.48004),but there was no significant difference between the two groups(t=1.114,p=0.338);3.The expression of Bcl-2 m RNA in the CMS group was no significant different with that in the control group(0.9205±0.14506 vs 1.0176±0.2666,t=0.738,p=0.427).4.The apoptosis rate in CMS group(CD71+ cells)was 2.4%,compared with the control group(2.3%),there was no significant different between the two groups(t= 0.409,p =0.686).5.There was no obvious relationship between Hb,apoptosis rate and Bax protein in CMS patients(p>0.05).Conclusion: 1.The expression of Bax m RNA in CMS group and control group was statistically different.The result showed that the pro-apoptotic gene Bax m RNA decreased in bone marrow erythroblasts of CMS patients.It was suggested that apoptosis may be involved in the pathogenesis of CMS disease;2.The expressions of Bcl-xl and Bcl-2 m RNA between the two groups were not statistically different,respectively.We need to increase the sample amount and continue to perform the relevant experiments to explore the effect of anti-apoptotic gene in the pathogenesis of CMS;3.There was no obvious relationship between Hb,apoptosis rate and Bax protein in CMS patients,It was suggested that cell proliferation played a major role possiblely.We need to increase the sample amount and continue to research. |
PDF Full Text Request