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目的:研究Ras/Raf/MEK/ERK信号通路蛋白激酶MEK特异性抑制剂PD98059对高原红细胞增多症(HAPC)患者骨髓CD71~+、CD235a~+有核红细胞增殖和凋亡影响及HAPC发病机制。方法:采用免疫磁珠法分选16例HAPC患者及16例对照者骨髓CD71~+、CD235a~+有核红细胞,分别加入5、10和20μmol/L不同浓度PD98059及DM SO溶剂,低氧培养72 h,以Annexin V和PI双染流式细胞术测定细胞凋亡,CCK8检测细胞增殖。同时观察加入5、10和20μmol/L等不同浓度PD98059及DMSO溶剂对骨髓单个核细胞(BMMNC)红系集落形成能力。结果:HAPC患者骨髓CD71~+、CD235a~+有核红细胞凋亡率随PD98059浓度增加上升(r=0.807,P<0.01);增殖率随PD98059浓度增加而下降(r=0.502,P<0.01)。HAPC患者骨髓BM M NC红系集落形成率随PD98059浓度增加而下降(r=0.504,P<0.01),7和14 d时各组比较均有统计学差异(P<0.05)。结论:M EK特异性抑制剂PD98059对HAPC患者CD71~+、CD235a~+有核红细胞具有抑制增殖、促进凋亡作用,对红细胞过度积累有一定的抑制作用。
Objective: To investigate the effect of PD98059, a specific MEK inhibitor of Ras / Raf / MEK / ERK signaling pathway, on the proliferation and apoptosis of CD71 +, CD235a ~ + erythroblasts in patients with high altitude polycythemia (HAPC) and the pathogenesis of HAPC. Methods: The bone marrow CD71 ~ +, CD235a ~ + nucleated erythrocytes from 16 patients with HAPC and 16 controls were collected by immunomagnetic beads method. The cells were treated with 5, 10 and 20 μmol / L PD98059 and DM SO respectively, 72 h, apoptosis was detected by Annexin V and PI double staining flow cytometry, and cell proliferation was detected by CCK8. At the same time, we observed the erythrocyte formation ability of bone marrow mononuclear cells (BMMNC) by adding PD98059 and DMSO with different concentrations such as 5,10 and 20μmol / L. Results: The apoptosis rate of CD71 +, CD235a ~ + nucleated erythrocytes in HAPC patients increased with the increase of PD98059 concentration (r = 0.807, P <0.01), and the proliferation rate decreased with the increase of PD98059 concentration (r = 0.502, . The erythrocyte formation rate of BMMNC in bone marrow of HAPC patients decreased with the increase of PD98059 concentration (r = 0.504, P <0.01), and there was a significant difference between 7 and 14 days (P <0.05). CONCLUSION: M EK specific inhibitor PD98059 can inhibit the proliferation and promote the apoptosis of CD71 +, CD235a ~ + nucleated erythrocytes in HAPC patients, and inhibit the excessive accumulation of erythrocytes.