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目的:探讨一氧化氮供体S-亚硝基谷胱甘肽(S-nitrosoglutathione,GSNO)对巨核细胞株meg-01诱导凋亡的可能作用。方法:将meg-01细胞株分为三组,对照组不加S-谷胱甘肽培养2 h,实验组分别加50μmol/L、100μmol/L、150μmol/L、200μmol/L的S-谷胱甘肽培养2 h,细胞因子组在含TPO(50 ng/ml)、IL-3(10 ng/ml)、SCF(50 ng/ml)的无血清培养基中培养72 h,再加入150μmol/LS-谷胱甘肽培养2 h。采用流式细胞仪检测凋亡细胞,电子显微镜观察细胞的形态学情况,实时荧光定量PCR检测Bcl2和Bax表达。结果:与对照组相比,实验组凋亡细胞数量明显增加,其中150μmol/L S-谷胱甘肽诱导凋亡细胞数量最多;电子显微镜下可见巨核细胞胞浆有明显空泡,细胞核固缩、破裂;实时荧光PCR检测到Bax的表达不同程度增加,Bcl2表达不同程度降低,其中150μmol/L GSNO诱导后,Bax的表达显著增加,Bcl2表达显著降低。细胞因子组Bax的表达降低,Bcl2表达增加。结论:S-谷胱甘肽可以促进巨核细胞株凋亡,而TPO、IL-3、SCF联合可抑制凋亡。
Objective: To investigate the possible role of nitric oxide donor S-nitrosoglutathione (GSNO) in inducing apoptosis of megakaryocyte meg-01 cells. Methods: The meg-01 cell lines were divided into three groups. The control group was incubated with S-glutathione for 2 h. The experimental groups were treated with 50 μmol / L, 100 μmol / L, 150 μmol / L, 200 μmol / The cytokines were cultured in serum-free medium containing TPO (50 ng / ml), IL-3 (10 ng / ml) and SCF (50 ng / ml) for 72 h after addition of 150 μmol / LS-glutathione for 2 h. Apoptotic cells were detected by flow cytometry. Morphology of the cells was observed by electron microscopy. The expression of Bcl2 and Bax was detected by real-time fluorescence quantitative PCR. Results: Compared with the control group, the number of apoptotic cells in the experimental group increased significantly, of which 150μmol / L S-glutathione induced the largest number of apoptotic cells; under the electron microscope, the cytoplasm of megakaryocytes showed obvious vacuoles and nuclear condensation , And the expression of Bax was increased to some extent by real-time fluorescence PCR. The expression of Bcl2 was decreased to different extents. After 150μmol / L GSNO was induced, the expression of Bax was significantly increased and the expression of Bcl2 was significantly decreased. Cytokines Bax expression decreased, Bcl2 expression increased. Conclusion: S-glutathione can promote the apoptosis of megakaryocytic cell line, and TPO, IL-3 and SCF can inhibit apoptosis.