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随着社会与科技的进步,核技术被广泛应用于工业、农业、医学和军事等领域。电离辐射会对机体造成急性、慢性或持久性损伤。造血系统因其对辐射的敏感性使其在辐射损伤的治疗中起着重要作用。造血生长因子(例如粒细胞集落刺激因子和粒细胞-巨噬细胞集落刺激因子)目前已获得美国食品和药物管理局的批准用于治疗造血系统的辐射损伤[1]。然而,应用造血生长因子不仅会导致发烧、疼痛和呕吐等,而且会破坏造血干细胞(hematopoietic stem cell, HSC)的自我更新能力,从而加速HSC的耗竭并进一步影响造血系统的长期恢复。因此,造血系统辐射损伤的治疗方法研究仍需进一步开展。
西格列汀(sitagliptin)是广泛用于临床治疗2型糖尿病的药物[2-4]。前期研究结果表明,西格列汀可以降低急性胰腺炎、糖尿病心肌病、慢性脑灌注不足、心力衰竭和肝缺血再灌注等疾病中的氧化应激水平[5-9]。Broxmeyer等[10]发现,提前给予小鼠西格列汀或敲除二肽基肽酶4(DPP4),可增强辐射暴露后机体的造血能力恢复。然而,西格列汀在造血系统辐射损伤中的治疗效果和机制尚待研究。
本研究在观察西格列汀对正常小鼠外周血血象是否有影响的基础上,探究了西格列汀在小鼠造血系统辐射损伤中的治疗效果,为寻找造血系统辐射损伤的潜在治疗药物研究打下基础。
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由表1可知,与对照组相比,照射组小鼠BMNC、外周血WBC、RBC、PLT计数和HGB、HCT水平均显著减少、降低,差异均有统计学意义(t=3.476~12.200,均P<0.05);与照射组相比,西格列汀+照射组小鼠的BMNC和WBC计数均显著增加,差异均有统计学意义(均P<0.05)。以上结果提示西格列汀可促进辐射暴露后的骨髓细胞及外周血的恢复。
组别 骨髓有核细胞
(×106个)白细胞
(×109个/L)红细胞
(×1012个/L)血小板
(×109个/L)血红蛋白
(g/L)红细胞比容
(%)对照组(n=5) 20.5±2.2 11.7±2.2 9.2±0.4 443.6±68.4 131.0±5.8 31.2±1.2 照射组(n=5) 8.0±1.0a 3.0±0.2a 7.9±0.6a 213.3±48.3a 117.6±6.4a 28.3±1.4a 西格列汀+照射组(n=5) 11.0±0.4ab 3.9±0.3ab 7.4±0.7a 362.0±66.7a 110.0±10.4a 26.8±2.0a 注:表中,a表示与对照组比较,t=2.510~12.200,均P<0.05;b表示与照射组比较,t=4.593、7.020,均P<0.05 表 1 西格列汀对7 Gy γ 射线全身照射后小鼠外周血血象及骨髓有核细胞的影响(
±s)$\bar x $ Table 1. Effect of sitagliptin on peripheral blood hemogram and bone marrow nucleated cells of mice exposed to 7 Gy γ-ray whole-body irradiation (
±s)$\bar x $ -
由图1可见,与对照组相比,照射组小鼠HSC和HPC的数量、百分比均显著减少、降低(均P<0.05),这表明电离辐射对HSC和HPC有一定的损伤作用;与照射组相比,西格列汀+照射组小鼠HPC的数量[(33 724.4±10 594.9)个对(101 637.6±17 240.5)个]、百分比[(5.6±1.0)%对(11.5±3.0)%]均增加,差异均有统计学意义(均P<0.05)。以上结果表明,西格列汀可缓解受照小鼠HPC和HSC的损伤,对辐射所致造血功能损伤有一定的治疗效果。
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与对照组相比,照射组、西格列汀+照射组小鼠骨髓细胞CFU-GM均显著减少,且差异均有统计学意义(t=13.240、9.436,均P<0.05);与照射组相比,西格列汀+照射组小鼠骨髓细胞CFU-GM增加,且差异有统计学意义(t=2.964,P<0.05)(CFU-GM:对照组66.2±5.3、照射组30.8±3.9、西格列汀+照射组38.2±4.4)。以上结果表明,西格列汀治疗可提高7 Gy照射小鼠骨髓细胞的克隆形成能力,一定程度上能够恢复骨髓细胞的增殖功能。
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由图2可见,与对照组相比,照射组小鼠在受照后第30天HSC中的活性氧水平显著升高(P<0.05);与照射组相比,西格列汀+照射组小鼠HSC 中的活性氧水平显著降低(P<0.05)(HSC中的活性氧水平:对照组1980.6±309.3、照射组3994.5±1119.0、西格列汀+照射组2415.7±122.9)。这些结果提示西格列汀对电离辐射所致的造血系统辐射损伤的保护作用可能是通过降低HSC中活性氧水平实现的。
西格列汀对小鼠造血系统辐射损伤的治疗作用
Therapeutic effect of sitagliptin on the radiation injury of the hematopoietic system in mice
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摘要:
目的 探讨西格列汀对7 Gy γ射线照射后小鼠造血系统损伤的治疗作用。 方法 将15只C57BL/6小鼠按照区组随机法分为对照组、照射组、西格列汀+照射组,每组5只。照射组和西格列汀+照射组小鼠接受7 Gy 137Cs γ射线一次性全身照射,后者于照射后2 h开始灌胃西格列汀,给药剂量为10 mg/kg,连续给药7 d;对照组和照射组给予等量生理盐水。照射后第30天取外周血进行血象测定,颈椎脱位处死小鼠后取骨髓细胞测定有核细胞数、造血干细胞(HSC)和造血祖细胞(HPC)数量及其活性氧水平、粒细胞-巨噬细胞集落形成单位(CFU-GM)。2组间的比较采用独立样本t检验。 结果 与对照组比较,照射组小鼠的骨髓有核细胞、白细胞、红细胞、血小板计数和血红蛋白、红细胞比容水平均明显减少、下降(t=3.476~12.200,均P<0.05);HSC和HPC的数量及百分比均明显减少、降低(t=3.174~5.287,均P<0.05);HSC中活性氧水平明显升高(1980.6±309.3对3994.5±1119.0,t=3.904,P<0.05),骨髓细胞CFU-GM显著减少(66.2±5.3对30.8±3.9,t=13.240,P<0.05)。与照射组相比,西格列汀+照射组小鼠骨髓有核细胞[(8.0±1.0)×106个对(11.0±0.4)×106个,t=4.593,P<0.05]、白细胞数量[(3.0±0.2)×109个/L对(3.9±0.3)×109个/L,t=7.020,P<0.05]均增多;HPC数量[(33 724.4±10 594.9)个对(101 637.6±17 240.5)个,t=5.951,P<0.05]及百分比[(5.6±1.0)%对(11.5±3.0)%,t=4.163,P<0.05]均上升,HSC中活性氧水平降低(3994.5±1119.0对2415.7±122.9,t=2.375,P<0.05),骨髓细胞CFU-GM增加(30.8±3.9对38.2±4.4,t=2.964,P<0.05)。 结论 西格列汀对7 Gy γ射线照射后小鼠造血系统损伤有一定的治疗作用。 Abstract:Objective To investigate the therapeutic effect of sitagliptin on hematopoietic system injury in mice after 7 Gy γ-ray irradiation. Methods Fifteen C57BL/6 mice were randomly divided into a control group, an irradiation group, and a sitagliptin+irradiation group by using a randomized block design, with five mice in each group. The mice in the irradiation and sitagliptin+irradiation groups received 7 Gy 137Cs γ-ray total body irradiation. Sitagliptin was administered to the latter via gavage 2 h after the irradiation. The dose was 10 mg/kg, and gavage infusion was continuous. Sitagliptin administration lasted for 7 days. The control and irradiation groups were given the same volume of normal saline. On the 30th day after irradiation, peripheral blood was collected for blood routine examination. After the mice were sacrificed via cervical dislocation, bone marrow cells were collected to determine the numbers of nucleated cells, hematopoietic stem cells (HSC), and hematopoietic progenitor cells (HPC), along with their reactive oxygen species (ROS) levels. The number of colony-forming units granulocyte-macrophage (CFU-GM) clones was also determined. Data between two groups were compared via independent sample t-test. Results Compared with those of the control group, the numbers of bone marrow nucleated cells, white blood cells, red blood cells, platelets, hemoglobin, and hematocrit of the irradiation group were significantly reduced (t=3.476−12.200, all P<0.05). The number and percentage of HSC and HPC were significantly reduced (t=3.174−5.287, all P<0.05). The level of ROS in HSC increased significantly (1980.6±309.3 vs. 3994.5±1119.0, t=3.904, P<0.05). The number of bone marrow cell CFU-GM clones was significantly decreased (66.2±5.3 vs. 30.8±3.9, t=13.240, P<0.05). Compared with the irradiation group, the number of bone marrow nucleated cells ((8.0±1.0)×106 vs. (11.0±0.4)×106, t=4.593, P<0.05) and white blood cells ((3.0±0.2)×109/L vs. (3.9±0.3)×109/L, t=7.020, P<0.05) increased (t=4.593, P<0.05) in the sitagliptin+irradiation group. The number of HPC (33 724.4±10 594.9 vs. 101 637.6±17 240.5, t=5.951, P<0.05) and percentage ((5.6±1.0%) vs. (11.5±3.0)%, t=4.163, P<0.05) also increased. The ROS level in HSC decreased (3994.5±1119.0 vs. 2415.7±122.9, t=2.375, P<0.05), whereas the number of the bone marrow cell CFU-GM clones increased (30.8±3.9 vs. 38.2±4.4, t=2.964, P<0.05). Conclusion Sitagliptin exerts a certain therapeutic effect on hematopoietic system injury in mice after 7 Gy γ-ray irradiation. -
表 1 西格列汀对7 Gy γ 射线全身照射后小鼠外周血血象及骨髓有核细胞的影响(
±s)$\bar x $ Table 1. Effect of sitagliptin on peripheral blood hemogram and bone marrow nucleated cells of mice exposed to 7 Gy γ-ray whole-body irradiation (
±s)$\bar x $ 组别 骨髓有核细胞
(×106个)白细胞
(×109个/L)红细胞
(×1012个/L)血小板
(×109个/L)血红蛋白
(g/L)红细胞比容
(%)对照组(n=5) 20.5±2.2 11.7±2.2 9.2±0.4 443.6±68.4 131.0±5.8 31.2±1.2 照射组(n=5) 8.0±1.0a 3.0±0.2a 7.9±0.6a 213.3±48.3a 117.6±6.4a 28.3±1.4a 西格列汀+照射组(n=5) 11.0±0.4ab 3.9±0.3ab 7.4±0.7a 362.0±66.7a 110.0±10.4a 26.8±2.0a 注:表中,a表示与对照组比较,t=2.510~12.200,均P<0.05;b表示与照射组比较,t=4.593、7.020,均P<0.05 -
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