| 张恂,张楠,郭艳茹,等.四物汤对模式生物斑马鱼血液系统与辐射损伤的影响[J].中华放射医学与防护杂志,2020,40(11):820-825.Zhang Xun,Zhang Nan,Guo Yanru,et al.Effect of Siwu decoction on the radiation injury of Zebrafish blood system[J].Chin J Radiol Med Prot,2020,40(11):820-825 |
| 四物汤对模式生物斑马鱼血液系统与辐射损伤的影响 |
| Effect of Siwu decoction on the radiation injury of Zebrafish blood system |
| 投稿时间：2020-04-21 |
| DOI：10.3760/cma.j.issn.0254-5098.2020.11.002 |
| 中文关键词: 斑马鱼 四物汤 血液系统 照射 |
| 英文关键词:Zebra fish Siwu decoction Blood system Irradiation |
| 基金项目:2020-09-30 |
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| 摘要点击次数: 2957 |
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| 中文摘要: |
| 目的 使用斑马鱼观察中药复方四物汤对辐射损伤血液系统的干预作用，明确斑马鱼是否可应用于中药对血液系统调控的研究。方法 72只体重在0.14~0.20 g的4月龄成年雄性斑马鱼（Danio rerio）用于全部实验，首先取36只斑马鱼用于照射后血液发育动力学的变化观察，另外36只斑马鱼用于四物汤对照射后血液系统的干预作用研究。采用60Co γ射线照射，辐射剂量20 Gy，吸收剂量率97.33 cGy/min。在照射后血液发育动力学观察中，9只未照射作为0 d对照，余27只斑马鱼分别于照射后第7、14和30天麻醉，收集外周血细胞和肾髓中细胞，采用流式细胞仪分析细胞的绝对数量。依据照射后外周血和肾髓中细胞的动态变化情况判断血液发育动力学特征，藉此实施四物汤的药效观察。四物汤对照射损伤干预实验周期7 d，36只斑马鱼采用随机数表法分为未照射组、单纯照射组、药物2 000组和药物5 000组，每组9只。两个药物组的斑马鱼按上述条件20 Gy照射后第2天分别置于稀释了2 000倍和5 000倍的四物汤水中饲养，第7天收集各组斑马鱼的外周血和肾髓中细胞，比较判断四物汤对斑马鱼辐射损伤血液系统的干预作用。结果 照射后斑马鱼血液发育动态变化明显，血液系统消融作用在第7天最明显，与未照射比较，斑马鱼的外周血细胞和肾髓总细胞数量分别下降26%和52%，差异均有统计学意义（t=4.535，28.987，P<0.05），肾髓中髓系单核细胞、淋巴细胞和红细胞数分别下降46%、79%和33%，差异均有统计学意义（t=18.457、66.900、9.872，P<0.05），前体细胞下降49%，但差异无统计学意义（P>0.05）。照射后第14天肾髓中各种细胞数均呈现上升，第30天恢复至照射前水平以上。斑马鱼照射后第2天开始连续6 d给予四物汤，药物5 000组与单纯照射组比较，肾髓细胞总数、髓系单核细胞、前体细胞和红细胞分别增加了57%、125%、81%和35%，差异均有统计学意义（t=12.128、21.594、15.473、4.594，P<0.05），淋巴细胞增加了59%，而外周血红细胞数量两组比较差异无统计学意义（P>0.05）；药物2 000组各参数均与单纯照射组比较差异无统计学意义（P>0.05）。结论 照射后斑马鱼血液系统发育呈现明显的先抑后扬的动态变化，一定浓度的四物汤对斑马鱼造血系统照射损伤有明显的干预作用，可为中药血液系统调控方面的研究提供参考。 |
| 英文摘要: |
| Objective To observe the effect of Chinese herbal compound Siwu decoction (SW) on radiation injury of zebra fish blood system, and to clarify whether zebra fish is a suitable model for this study. Methods Totally 72 of 4-month-old adult male zebra fish (Danio Rerio) with body weight ranging from 0.14 to 0.20 g were used in all the experiments, where 36 zebra fish were taken to observe the hemodynamic changes after radiation, and the other 36 zebra fish were used to study the intervention effect of Siwu decoction on the blood injury after radiation. Zebra fish was irradiated with 20 Gy of 60Co gamma-rays at a dose rate of 97.33 cGy/min. For the hemodynamic study, the peripheral blood and whole kidney marrow cells (WKMCs)of irradiated zebra fish and non irradiated control were collected on days 7, 14 and 30 after irradiation, respectively, and the cell number was detected by flow cytometry. For the Siwu decoction treatment, 36 zebra fish were randomly divided into non-irradiation group, irradiation group, SW2 000 group (Siwu decoction was diluted by 2 000-fold) and SW5 000 group (Siwu decoction was diluted by 5 000-fold). Siwu decoction was given on the second day after irradiation. Then the peripheral blood and WKM cells of zebra fish were collected at 7 d after irradiation for further analysis. ResultsAfter irradiation, the dynamic of blood development of zebra fish were obviously changed with the most blood cell ablation at 7 d post-irradiation. The total number of peripheral blood cells and WKM cells decreased by 26% and 52%(t=4.535, 28.987, P<0.05), the number of myeloid monocytes, lymphocytes, and red blood cells in the WKM decreased by 46%, 79% and 33%(t=18.457, 66.900, 9.872, P<0.05), and the number of precursor cells decreased by 49% (P>0.05). The numbers of these blood cells were partly recovered at 14 d after irradiation and returned to the normal level at 30 d after irradiation. In comparison with irradiation alone group, the numbers of WKM cells, myeloid monocytes, progenitor cells, lymphocytes and red blood cells increased by 57%,125%, 81% and 35%, respectively (t=12.128, 21.594, 15.473, 4.594,P<0.05),and peripheral red cells did not increase significantly in the SW5 000 group after irradiation. However, SW2 000 had no significant influence on radiation injury of zebra fish blood cells (P>0.05). Conclusions Radiation injury of the blood system of zebra fish could be recovered at a month after irradiation, and this radiation damage could be partly intervened by Siwu decoction at a suitable concentration, which provides a reference for the study of blood system regulation of traditional Chinese medicine. |
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