低氧诱导因子1α在G-CSF诱导小鼠造血干细胞动员中的表达变化  

Expression Changes of Hypoxia-Inducible Factor-1α in G-CSF Induced Hematopoietic Stem Cell Mobilization

作　　者：杨慧璇 李桥川[1] 韦莉莉[2] 赖永榕[1] YANG Hui-Xuan;LI Qiao-Chuan;WEI Li-Li;LAI Yong-Rong(Department of Hematology,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;General Geriatrics Division,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第一附属医院血液内科,广西南宁530021 [2]广西医科大学第一附属医院老年病学综合病区,广西南宁530021

出　　处：《中国实验血液学杂志》2023年第1期221-226,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81960038)。

摘　　要：目的:探究小鼠骨髓微环境低氧诱导因子1α(HIF-1α)在G-CSF动员造血干细胞(HSC)过程中的表达变化及相关机制。方法:采用流式细胞术检测注射G-CSF前后小鼠外周血Lin-Sca-1+c-kit+(LSK)细胞比例,并采用RQ-PCR、免疫组化等方法检测G-CSF动员不同时间点小鼠骨髓及骨内膜HIF-1α、骨钙素(OCN)mRNA和蛋白的表达水平,光学显微镜下观察小鼠股骨标本成骨细胞数。结果:G-CSF动员4 d小鼠外周血LSK细胞比例开始升高,动员5 d达到高峰,均较对照组显著升高(P<0.05);稳态小鼠HIF-1α mRNA在骨髓有核细胞和骨内膜成骨细胞中表达差异无显著统计学意义(P=0.073),OCN mRNA主要表达于骨内膜细胞,显著高于骨髓有核细胞(P=0.034);动员过程中HIF-1α基因、蛋白表达水平升高,OCN基因、蛋白表达水平下降,骨内膜成骨细胞数量减少(P<0.05);HIF-1α的表达变化晚于OCN(P=0.004),且与小鼠外周血LSK细胞比例变化趋势一致(P=0.000)。结论:在G-CSF介导的HSC动员过程中,骨髓HIF-1α表达增加,成骨细胞受抑引起HSC向外周迁移是导致其变化的原因之一。Objective: To investigate the expression and its relative mechanism of hypoxia-inducible factor-1α(HIF-1α) in bone marrow(BM) of mice during G-CSF mobilization of hematopoietic stem cells(HSC). Methods: Flow cytometry was used to detect the proportion of Lin-Sca-1+c-kit+(LSK) cells in peripheral blood of C57 BL/6 J mice before and after G-CSF mobilization. And the expression of HIF-1α and osteocalcin(OCN) mRNA and protein were detected by RQ-PCR and immunohistochemistry. The number of osteoblasts in bone marrow specimens of mice was counted under the microscope. Results: The proportion of LSK cells in peripheral blood began to increase at day 4 of G-CSF mobilization, and reached the peak at day 5, which was significantly higher than that of control group(P<0.05). There was no distinct difference in the expression of HIF-1α mRNA between bone marrow nucleated cells and osteoblasts of steady-state mice(P=0.073), while OCN mRNA was mainly expressed in osteoblasts, which was higher than that in bone marrow nucleated cells(P=0.034). After mobilization, the expression level of HIF-1α increased, but OCN decreased, and the number of endosteum osteoblasts decreased. The change of HIF-1α expression was later than that of OCN and was consistent with the proportion of LSK cells in peripheral blood. Conclusion: The expression of HIF-1α in bone marrow was increased during the mobilization of HSC mediated by G-CSF, and one of the mechanisms may be related to the peripheral migration of HSC induced by osteoblasts inhibition.

关 键 词：低氧诱导因子1Α 成骨细胞 造血干细胞动员 骨髓微环境 

分 类 号：R457.7[医药卫生—治疗学]