Yang L Y, Wang H Q, Fu R, Qu W, Ruan E B, Wang X M, Wang G J, Wu Y H, Liu H, Song J, Guan J, Xing L M, Li L J, Jiang H J, Liu H, Wang Y H, Liu C Y, Zhang W, Shao Z H
Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
Zhonghua Xue Ye Xue Za Zhi. 2017 May 14;38(5):432-436. doi: 10.3760/cma.j.issn.0253-2727.2017.05.015.
To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) . Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA(+) nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA(+) nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA(+) nucleated RBC was detected by Western blot. Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA(+) nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, <0.001) , and lower than that in low-risk MDS patients (0.40±0.16, =0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA(+) nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , <0.017) . The expression of ULK1 mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , <0.017]. The level of mTOR mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , <0.017]. The level of LC3B in GlycoA(+) nucleated RBC was negatively correlated with the HGB (=0.529, =0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42. Autophagy is impaired in nucleated RBC of MDS patients.
探讨骨髓增生异常综合征（MDS）患者骨髓有核红细胞自噬水平的变化。本研究纳入了54例MDS患者和33例对照。通过透射电子显微镜（TEM）观察线粒体自噬。采用流式细胞术检测糖基化A（GlycoA）阳性有核红细胞中自噬相关蛋白LC3B的水平。采用实时聚合酶链反应（PCR）检测GlycoA阳性有核红细胞中ULK1和mTOR mRNA的表达。通过蛋白质免疫印迹法检测GlycoA阳性有核红细胞中线粒体外膜蛋白TOM20的表达。TEM观察发现MDS患者中几乎未观察到自噬体或自噬溶酶体。高危MDS患者GlycoA阳性有核红细胞中LC3B的表达（0.22±0.12）显著低于正常对照（0.43±0.22，P<0.001），且低于低危MDS患者（0.40±0.16，P=0.001）。高危MDS患者GlycoA阳性有核红细胞中AMPK的表达[0.26（0.60）]显著低于对照[1.00（2.07），P<0.017]。高危MDS患者GlycoA阳性有核红细胞中ULK1 mRNA的表达[0.27（3.31）]显著低于对照[1.07（4.41），P<0.017]。高危MDS患者GlycoA阳性有核红细胞中mTOR mRNA的水平[1.82（3.74）]显著高于对照[1.26（1.38），P<0.017]。高危MDS患者GlycoA阳性有核红细胞中LC3B的水平与血红蛋白（r=-0.529，P=0.009）呈负相关。高危MDS患者线粒体外膜蛋白TOM20的表达为9.42±4.42。MDS患者有核红细胞的自噬功能受损。
