| Sun Kangfu,Yang Lijing,Fu Zeyu,Xu Baichuan,Chen Shilei,Wang Junping.Effects of sleep disorders on hematopoietic stem cells in bone marrow of irradiated mice[J].Chinese Journal of Radiological Medicine and Protection,2024,44(2):96-104 |
| Effects of sleep disorders on hematopoietic stem cells in bone marrow of irradiated mice |
| Received:November 19, 2023 |
| DOI：10.3760/cma.j.cn112271-20231119-00177 |
| KeyWords:Sleep disorders Ionizing radiation Hematopoietic stem cell NOX2 Caspase-1 |
| FundProject:国家自然科学基金面上项目(82222060,82073487);重庆市研究生教育创新计划(CYS22742) |
| Author Name | Affiliation | E-mail | | Sun Kangfu | State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University(Third Military Medical University), Chongqing 400038, China | | | Yang Lijing | State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University(Third Military Medical University), Chongqing 400038, China | | | Fu Zeyu | State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University(Third Military Medical University), Chongqing 400038, China | | | Xu Baichuan | State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University(Third Military Medical University), Chongqing 400038, China | | | Chen Shilei | State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University(Third Military Medical University), Chongqing 400038, China | | | Wang Junping | State Key Laboratory of Trauma and Chemical Poisoning, Institute of Combined Injury, Faculty of Military Preventive Medicine, College of Basic Medical Sciences, Army Medical University(Third Military Medical University), Chongqing 400038, China | wangjunping@tmmu.edu.cn |
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| Abstract:: |
| Objective To investigate the effects of sleep disorders (SD) on the radiation injury of hematopoietic stem cells (HSCs) in bone marrow (BM). Methods Totolly 56 C57BL/6J male mice aged 6-8 weeks were enrolled in this study. They were subjected to whole body irradiation of 60Co γ-rays with doses of 5.0 and 7.5 Gy. A SD model was established using a SD device. According to the random number table method, the mice were divided into seven groups: the control group (Con group), the SD group, the mere radiation group (IR group), the group of post-irradiation SD (IR+SD group), the group of post-irradiation SD treated with phosphate buffer solution (IR+SD+PBS group), the group of post-irradiation SD treated with GSK2795039 (IR+SD+GSK group), and the group of post-irradiation SD treated with N-acetylcysteine (IR+SD+NAC group), with in eight mice each group. The changes in the peripheral blood of the mice after 5.0 Gy irradiation were detected using the collected tail venous blood, and the survival rates of the mice after 7.5 Gy irradiation were observed. The changes in the density and count of bone marrow cells were observed using hematoxylin and eosin (HE) staining. The number of hematopoietic stem cells in bone marrow (LSK cells), as well as their apoptosis level and changes in cell cycle, were detected using flow cytometry. Furthermore, indicators of LSK, such as reactive oxygen species(ROS) and mitochondrial-derived reactive oxygen species (mtROS), were analyzed. Nicotinamide adenine dinucleotide phosphate (NADP+/NADPH) and glutathione (GSSG/GSH) were detected using an enzyme microplate reader in order to observe the oxidative stress level of LSK. Furthermore, flow cytometry was employed to sort the LSK cells from the mice, and flow cytometry was used to detect the expression of NADPH oxidase 2(NOX2) and cysteinyl aspartate specific proteinnase-1(Caspase-1), and polymerase chain reaction (PCR) was used to detect the expression of inflammatory factors such as NOX1-4, interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 18 (IL-18), and tumor necrosis factor α (TNF-α). Results Compared to the IR group, the IR+SD group exhibited significantly slower recovery of white blood cells (WBC) and platelets (PLT) (t = 4.39, 6.37, P ＜ 0.05), the bone marrow cell count decreasing from (2.14 ± 0.38) × 107 to (3.59 ± 0.29) × 107 (t = 8.55, P ＜ 0.05), significantly decreased proportion of G0-phase LSK cells, significantly increased proportion of apoptotic cells (t = 7.53, 8.21, P ＜ 0.05), and significantly increased DCFH-DA, MitoSOX, and NADP+/NADPH (t = 22.99, 29.47, 3.77, P＜0.05). In the case of IR, SD further promoted the activation of NOX2 and led to increases in the mRNA expression of downstream inflammatory factors such as IL-1β, IL-6, IL-18, and TNF-α (t = 6.95, 6.01, 8.39, 4.91, 5.56, P ＜ 0.05). Inhibition of NOX2-ROS could prevent the SD-induced aggravation of post-irradiation hematopoietic injury. This significantly reduced the apoptotic rate of LSK cells and the expression of inflammatory factors, ultimately accelerating the hematopoietic recovery of LSK cells (t = 9.24, 3.92, P ＜ 0.05). Conclusions SD can aggravate the IR-induced injury of hematopoietic stem cells in bone marrow, primarily by activating the NOX2-ROS-Caspase-1 axis. This will increase the levels of intracellular inflammatory factors and ROS, promote cell apoptosis, and ultimately inhibit the hematopoietic recovery of bone marrow. |
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