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人体的造血系统和免疫系统对电离辐射极为敏感,机体受到电离辐射会引起造血及免疫系统的损伤,形成急性放射病。而随着核技术的发展和广泛应用,人类受到辐射的概率也在不断增加[1]。现批准临床应用的辐射防护药物仅氨磷汀一种,且必须静脉注射,应用存在很大限制。因此,开展新的具有辐射防护功效的药物研究意义重大,将为核技术的广泛应用提供安全保障。
本研究的前期实验是应用药物组合原理,在褪黑素主要结构功能单位的氨基位置引入具有抗自由基、抗辐射及免疫活性功效的基团α-硫辛酸,得到新化合物5-甲氧基色胺-α-硫辛酸盐(5-methoxytryptamine-α-lipoic acid salt,MLA)。初步研究发现,MLA对急性辐射损伤具有一定的防护作用[2]。本研究进一步探讨MLA对小鼠受照后30 d的辐射防护效果及其作用机制。
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小鼠接受6.0 Gy照射后30 d,外周血WBC计数检测结果见表 1。与对照组相比,照射组和照射+MLA组小鼠的WBC(t=10.68、8.45,均P<0.05)、RBC(t=6.437、5.819,均P<0.05)等计数结果均明显下降,差异具有统计学意义。与照射组比较,MLA可显著提高受照小鼠WBC和RBC等计数结果(t=4.858、3.335,均P<0.05),差异具有统计学意义。
组别 WBC/(×109/L) RBC/(×1012/L) 血红蛋白/(g/L) 红细胞比容/% 血小板/(×109/L) 对照组 7.6±1.8 8.4±0.3 112.0±7.0 28.5±1.7 379.0±84.0 照射组 2.5±0.3 7.0±0.6 90.0±11.0 24.6±1.9 337.0±50.0 照射+MLA组 3.5±0.3 8.2±0.2 110.0±6.0 27.8±1.3 517.0±186.0 注:表中,MLA:5-甲氧基色胺-α-硫辛酸盐。 表 1 6.0 Gy全身照射后30 d小鼠外周血计数(n=5)
Table 1. The peripheral blood counts of mice exposed to 6.0 Gy total body irradiation for 30 d(n=5)
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对小鼠单侧BMNCs计数及集落形成能力进行检测,结果见表 2。在小鼠接受6.0 Gy照射30 d后,与对照组相比,照射组BMNCs计数下降36%,集落形成能力下降58%,差异均具有统计学意义(t=9.304、7.493,均P<0.05)。与照射组比较,在给予MLA处理后,BMNCs计数和集落形成能力均有所增加,且差异具有统计学意义(t=5.629、3.947,均P<0.05)。
组别 BMNCs/(×106/股骨) CFU-GM/(×105细胞) 对照组 24.4±3.0 251.0±57.0 照射组 15.7±0.9 109.0±21.0 照射+MLA组 18.8±0.6 169.0±22.0 注:表中,MLA:5-甲氧基色胺-α-硫辛酸盐;BMNCs:骨髓有核细胞;CFU-GM:粒细胞-单核细胞集落生成数量。 表 2 6.0 Gy全身照射后30 d小鼠免疫器官指数的 变化(n=5)
Table 2. The immune organs index changes of mice exposed to 6.0 Gy total body irradiation for 30 d(n=5)
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对小鼠造血细胞活性氧水平进行检测分析,结果见图 1。与对照组比较,照射组小鼠BMNCs、造血祖细胞(hematopoietic progenitor cells,HPCs)和造血干细胞(hematopoietic stem cells,HSCs)中的活性氧水平均升高(t=14.74、41.77、20.04,均P<0.05)。与照射组比较,给予MLA处理后,3类细胞中的活性氧水平均出现显著下降,且差异具有统计学意义(t=11.21、26.80、4.968,均P<0.05)。
图 1 MLA对6.0 Gy照射后30 d小鼠造血细胞活性氧水平的影响。图中,A:MLA对骨髓有核细胞活性氧的影响;B:MLA对造血祖细胞活性氧的影响;C:MLA对造血干细胞活性氧的影响。 MLA:5-甲氧基色胺-α-硫辛酸盐;BMNCs:骨髓有核细胞;MFI:平均荧光强度;HPCS:造血祖细胞;HSCs:造血干细胞。
Figure 1. The effects of 5-methoxytryptamine-α-lipoic acid salt on the ROS levels in the hematopoietic cells of mice exposed to 6.0 Gy total body irradiation for 30 d
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对小鼠造血细胞NOX4表达进行检测,结果见图 2。与对照组比较,照射组小鼠BMNCs、HPCs和HSCs中的NOX4表达均升高,且差异有统计学意义(t=53.12、21.5、64.43,均P<0.05)。与照射组比较,给予MLA处理后,3类细胞中的NOX4表达均出现显著下降,且差异有统计学意义(t=33.93、13.83、12.93,均P<0.05)。
图 2 MLA对6.0 Gy照射后30 d小鼠造血细胞NOX4表达的影响。 图中,A:MLA对BMNCs NOX4表达的影响;B:MLA对HPCs NOX4表达的影响;C:MLA对HSCs NOX4表达的影响。MLA:5-甲氧基色胺-α-硫辛酸盐;BMNCs:骨髓有核细胞;NOX4:烟酰胺腺嘌呤二核苷磷酸氧化酶4;HPCs:造血祖细胞;HSCs:造血干细胞;MFI:平均荧光强度。
Figure 2. The effects of MLA on the NOX4 expression in the hematopoietic cells of mice exposed to 6.0 Gy total body irradiation for 30 d
5-甲氧基色胺-α-硫辛酸盐对6.0 Gy受照小鼠造血系统的辐射防护作用
Protective effects of 5-methoxytryptamine-α-lipoic acid salt on mice exposed to 6.0 Gy total body irradiation
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摘要:
目的 探讨新化合物5-甲氧基色胺-α-硫辛酸盐(MLA)对亚致死剂量受照小鼠造血系统损伤的辐射防护作用。 方法 将15只C57BL/6小鼠完全随机分为对照组、照射组和照射+MLA组。对照组接受假照射(0 Gy),其余两组进行6.0 Gy全身137Cs γ射线照射。照射后2 h将照射+MLA组小鼠按10 mg/kg灌胃给药,持续给药3 d。待照射后30 d处死小鼠,取外周血和单侧骨髓细胞进行计数,检测骨髓细胞克隆形成能力、造血细胞活性氧以及烟酰胺腺嘌呤二核苷磷酸氧化酶4(NOX4)的表达。 结果 与对照组比较,照射组小鼠骨髓有核细胞计数、粒细胞-单核细胞集落生成数量(CFU-GM)均明显下降(t=9.304、7.493,均P<0.05);骨髓细胞活性氧水平和NOX4表达显著升高(t=14.74、53.12,均P<0.05),且差异有统计学意义。与照射组相比,照射+MLA组小鼠外周血WBC、CFU-GM显著升高(t=4.858、3.947,均P<0.05);骨髓细胞活性氧水平和NOX4表达显著下降(t=11.21、33.93,均P<0.05),且差异有统计学意义。 结论 MLA对辐射引起的造血系统损伤有一定的防护作用。 -
关键词:
- 辐射防护 /
- 辐射损伤 /
- 活性氧 /
- NADPH氧化酶4 /
- 5-甲氧基色胺-α-硫辛酸盐
Abstract:Objective To study the protective effects of 5-methoxytryptamine-α-lipoic acid salt(MLA), a new synthetized compound, on the hematopoietic system of mice exposed to sublethal dose of total body irradiation(TBI). Methods Fifteen C57BL/6 mice were randomly assigned into three groups:control, irradiation, and MLA+irradiation. The mice in the control group received sham TBI, whereas the mice in the other groups underwent exposure to 6.0 Gy 137Cs γ-ray TBI. The mice in the MLA group were first administered with 10 mg/kg MLA after exposure to 6.0 Gy TBI for 2 h. The MLA was administered successively for 3 d. The peripheral blood and bone marrow nucleated cells(BMNCs) were then counted after the mice were exposed to TBI for 30 d. The CFU-GM ability, reactive oxygen species(ROS) levels, and nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4) expression in the hematopoietic cells were also detected. Results In contrast to those of the control group, the number of BMNCs and the number CFU-GM in the irradiated groups decreased significantly(t=9.304, 7.493, both P<0.05), and the ROS levels and the NOX4 expression in the bone marrow cells of the irradiated groups increased(t=14.74, 53.12, both P<0.05). Compared with those of the irradiation group, the number of white blood cells and the number of BMNCs were elevated by MLA(t=4.858, 3.947, both P<0.05). The ROS levels and NOX4 expression in the bone marrow cells were inhibited by MLA(t=11.21, 33.93, both P<0.05). Conclusion MLA can protect mice from hematopoietic injury induced by irradiation. -
图 1 MLA对6.0 Gy照射后30 d小鼠造血细胞活性氧水平的影响。图中,A:MLA对骨髓有核细胞活性氧的影响;B:MLA对造血祖细胞活性氧的影响;C:MLA对造血干细胞活性氧的影响。 MLA:5-甲氧基色胺-α-硫辛酸盐;BMNCs:骨髓有核细胞;MFI:平均荧光强度;HPCS:造血祖细胞;HSCs:造血干细胞。
Figure 1. The effects of 5-methoxytryptamine-α-lipoic acid salt on the ROS levels in the hematopoietic cells of mice exposed to 6.0 Gy total body irradiation for 30 d
图 2 MLA对6.0 Gy照射后30 d小鼠造血细胞NOX4表达的影响。 图中,A:MLA对BMNCs NOX4表达的影响;B:MLA对HPCs NOX4表达的影响;C:MLA对HSCs NOX4表达的影响。MLA:5-甲氧基色胺-α-硫辛酸盐;BMNCs:骨髓有核细胞;NOX4:烟酰胺腺嘌呤二核苷磷酸氧化酶4;HPCs:造血祖细胞;HSCs:造血干细胞;MFI:平均荧光强度。
Figure 2. The effects of MLA on the NOX4 expression in the hematopoietic cells of mice exposed to 6.0 Gy total body irradiation for 30 d
表 1 6.0 Gy全身照射后30 d小鼠外周血计数(n=5)
Table 1. The peripheral blood counts of mice exposed to 6.0 Gy total body irradiation for 30 d(n=5)
组别 WBC/(×109/L) RBC/(×1012/L) 血红蛋白/(g/L) 红细胞比容/% 血小板/(×109/L) 对照组 7.6±1.8 8.4±0.3 112.0±7.0 28.5±1.7 379.0±84.0 照射组 2.5±0.3 7.0±0.6 90.0±11.0 24.6±1.9 337.0±50.0 照射+MLA组 3.5±0.3 8.2±0.2 110.0±6.0 27.8±1.3 517.0±186.0 注:表中,MLA:5-甲氧基色胺-α-硫辛酸盐。 表 2 6.0 Gy全身照射后30 d小鼠免疫器官指数的 变化(n=5)
Table 2. The immune organs index changes of mice exposed to 6.0 Gy total body irradiation for 30 d(n=5)
组别 BMNCs/(×106/股骨) CFU-GM/(×105细胞) 对照组 24.4±3.0 251.0±57.0 照射组 15.7±0.9 109.0±21.0 照射+MLA组 18.8±0.6 169.0±22.0 注:表中,MLA:5-甲氧基色胺-α-硫辛酸盐;BMNCs:骨髓有核细胞;CFU-GM:粒细胞-单核细胞集落生成数量。 -
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