自噬
免疫印迹
液泡
巨核细胞
免疫荧光
化学
细胞生物学
ATG5型
分子生物学
生物
男科
免疫学
细胞凋亡
抗体
医学
细胞质
生物化学
基因
干细胞
造血
作者
Qi Wang,Li Yang,Tao Feng,Xiao Feng,Zheng-hua Ji,Xue-qiang Ji,Xuejun Shao
出处
期刊:Zhongguo shi yan xue ye xue za zhi
日期:2021-10-01
卷期号:29 (5): 1577-1581
标识
DOI:10.19746/j.cnki.issn.1009-2137.2021.05.032
摘要
OBJECTIVE To investigate the effect of enhanced autophagy in megakaryocyte to proplatelet formation in children with immune thrombocytopenia(ITP). METHODS Giemsa staining and immunofluorescence staining were used to observe megakaryocyte morphology and proplatelet formation, Western blot was used to determine the expression of cytoskeleton protein and autophagy related protein. Autophagr regulation drugs Rap or 3-MA was used to regulate autophagy of megakaryocytes. RESULTS Some vacuole-like structures was found in ITP megakaryocytes of the children, the expression of LC3II/I (ITP 1.32±0.18; Ctrl 0.49±0.16,P<0.05) and Atg5-Atg12 (ITP 0.69±0.17; Ctrl 0.12±0.08,P<0.05) was significantly higher in ITP children as compared with those in control group. The immu- nofluorescence staining showed that the cytoskeleton arrangement in megakaryocytes of ITP children was abnormal, and the phosphorylation of myosin light chain was also increased(ITP 0.74±0.09, Ctrl 0.05±0.02,P<0.05). In vitro, inducer or inhibitor of autophagy could regulate the production of proplatelet and the expression of cell cycle related protein, including CyclinD1(Veh 1.08±0.12; Rap 0.46±0.04; Rap+3-MA 0.70±0.03), CyclinD2(Veh 0.47±0.04; Rap 0.27±0.04; Rap+3-MA 0.41±0.03), P21(Veh 0.15±0.01; Rap 0.04±0.01; Rap+3-MA 0.05±0.01). CONCLUSION Enhanced autophagy is the key factor of poor proplatelet formation in megakaryocytes of ITP children.
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