低氧习服在小鼠造血干细胞急性放射损伤修复中的作用观察  

Observation of effect on hypoxia acclimatization in repair of hematopoietic stem cells after acute radiation injury in mice

作　　者：全勇 冉新泽[1] 胡梦佳 陈芳[1] 陈乃成 廖伟年 陈默[1] 申明强[1] 陈石磊[1] 王崧[1] 王军平[1] Quan Yong;Ran Xinze;Hu Meng jia;Chen Fang;Chen Naicheng;Liao Weinian;Chen Mo;Shen Mingqiang;Chen Shilei;Wang Song;Wang Junping(Institute of Combined Injury of PLA,State Key Laboratory of Trauma,Burn and Combined Injury,College of Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing Engineering Research Center of Nanomedicine,Chongqing 400038,China)

机构地区：[1]陆军军医大学(第三军医大学)军事预防医学系全军复合伤研究所创伤、烧伤与复合伤国家重点实验室重庆市纳米医学工程研究中心,重庆400038

出　　处：《中华损伤与修复杂志（电子版）》2023年第4期293-298,共6页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：国家自然科学基金重点项目(81930090);军队后勤科研项目(ALJ19J002)。

摘　　要：目的研究低氧习服在小鼠造血干细胞急性放射损伤后修复中的作用。方法采用随机数字表法将6~7周龄雄性C57BL/6J小鼠分为常氧辐照组和低氧习服辐照组。建立低氧习服急性放射损伤动物模型,低氧习服后再进行60 Co全身一次性照射6.0Gy。连续监测辐照前后小鼠尾静脉血血常规;流式细胞术检测低氧习服前后小鼠骨髓造血干细胞;取低氧习服小鼠长骨进行病理分析和VE-Cadherin免疫荧光染色;ELISA检测低氧习服小鼠低氧诱导因子-1α(HIF-1α)和血管生成素-1(ANGPT1)。结果低氧习服加速辐照后小鼠外周血恢复,低氧习服辐照组外周血红细胞和血小板最低值高于常氧辐照组[(7.0±0.6)×10^(12)/L vs(5.1±0.4)×10^(12)/L,t=2.483,P<0.05;(203±30)×10^(9)/L vs(141±25)×10^(9)/L,t=2.541,P<0.05];低氧习服组骨髓LSK细胞和长期造血干细胞(LT-HSC)的比例及数量均高于常氧对照组[(3.91±0.21)‰vs(3.37±0.11)‰,t=2.536,P=0.032;(7.18±0.84)×10^(4)/L vs(6.01±0.82)×10^(4)/L,t=2.641,P=0.029;(12.05±0.53)%vs(9.37±0.45)%,t=3.512,P=0.023;(0.74±0.12)×10^(3)/L vs(0.58±0.11)×10^(3)/L,t=2.667,P=0.027]。低氧习服促进骨髓窦的增殖扩张和VE-Cadherin在窦内皮细胞的表达,减轻造血细胞凋亡和血管龛损伤;低氧习服组小鼠骨髓LSK细胞中HIF-1α和骨髓腔冲洗上清液中ANGPT1含量明显增加[(2.97±0.28)pg/cell vs(1.95±0.22)pg/cell,t=4.896,P=0.013;(12.30±3.70)pg/ml vs(6.25±2.70)pg/ml,t=6.742,P=0.001]。结论低氧习服减轻造血干细胞和血管龛的放射损伤,加快造血恢复,其原因可能与低氧习服调节LT-HSC比例,促进HIF-1α、ANGPT1在造血干细胞中的表达有关。Objective To investigate the effect of hypoxia acclimatization in repair of hematopoietic stem cells(HSCs)after acute radiation injury in mice.Methods According to random number table method,the male C57BL mice aged 6-7 weeks were divided into normoxic irradiation group and hypoxia acclimatization irradiation group.The animal model of acute radiation injury after hypoxia acclimatization was established by 6.0 Gy of one-time irradiation of 60 Co.Tail venous blood routine of mice before and after irradiation was continuously monitored.Detected the bone marrow hematopoietic stem cells in mice before and after hypoxia acclimatization by flow cytometry.Long bones of hypoxic acclimatized mice were taken for pathological analysis and VE-Cadherin immunofluorescence staining.HIF-1αand ANGPT1 were detected by ELISA in hypoxic acclimatized mice.Results Hypoxia acclimatization accelerated the recovery of peripheral blood after irradiation,the lowest numbers of RBC and PLT in hypoxia acclimatization irradiation group were higher than normoxic irradiation group[(7.0±0.6)×10^(12)/L vs(5.1±0.4)×10^(12)/L,t=2.483,P<0.05;(203±30)×10^(9)/L vs(141±25)×10^(9)/L,t=2.541,P<0.05].The proportions and numbers of LSK cells and LT-HSC in the bone marrow of hypoxia acclimatization group were significantly higher than normoxic control group[(3.91±0.21)‰vs(3.37±0.11)‰,t=2.536,P=0.032;(7.18±0.84)×10^(4)/L vs(6.01±0.82)×10^(4)/L,t=2.641,P=0.029;(12.05±0.53)%vs(9.37±0.45)%,t=3.512,P=0.023;(0.74±0.12)×10^(3)/L vs(0.58±0.11)×10^(3)/L,t=2.667,P=0.027].Hypoxia acclimatization caused proliferation and expansion of the bone marrow sinuses,and strengthened the expression of VE-Cadherin in endothelial sinusoidal cells.It also alleviated hematopoietic cell apoptosis and vascular niche damage.HIF-1αin LSK cells and ANGPT1 in the bone marrow cavity flushing supernatant increased significantly[(2.97±0.28)pg/cell vs(1.95±0.22)pg/cell,t=4.896,P=0.013;(12.30±3.70)pg/ml vs(6.25±2.70)pg/ml,t=6.742,P=0.001].Conclusion Hypoxia acclimat

关 键 词：低氧习服 放射损伤 造血干细胞 低氧诱导因子-1Α 血管生成素-1 

分 类 号：R644[医药卫生—外科学]