基础医学与临床 ›› 2023, Vol. 43 ›› Issue (2): 233-240.doi: 10.16352/j.issn.1001-6325.2023.02.233

• 研究论文 • 上一篇    下一篇

AHR抑制剂CH223191提高辐照小鼠骨髓移植效果

庞瑞洋, 周力, 吕家迪*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 免疫学系 医学分子生物学国家重点实验室,北京 100005
  • 收稿日期:2022-10-11 修回日期:2022-11-22 出版日期:2023-02-05 发布日期:2023-02-02
  • 通讯作者: *lvjiadi666@aliyun.com
  • 基金资助:
    国家自然科学基金(82003145)

AHR inhibitor CH223191 improves the effect of bone marrow transplantation in irradiated mice

PANG Ruiyang, ZHOU Li, LYU Jiadi*   

  1. National Key Laboratory of Medical Molecular Biology, Department of Immunology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005,China
  • Received:2022-10-11 Revised:2022-11-22 Online:2023-02-05 Published:2023-02-02
  • Contact: *lvjiadi666@aliyun.com

摘要: 目的 探究骨髓移植过程中辐射诱导的旁观者效应对供体骨髓细胞的影响以及芳香烃受体(AHR)抑制剂CH223191对供体骨髓细胞的保护作用。方法 将小鼠分为9 Gy辐照组和12 Gy辐照组,不同辐照剂量辐射后移植相同数量供体骨髓细胞,流式细胞术检测外周血细胞总数、外周血供体细胞嵌合率及数量;ELISA检测未辐照小鼠和辐照小鼠血清活性氧(ROS)水平;将供体骨髓细胞分为对照组、9 Gy辐照组和12 Gy辐照组体外培养,分别加入未辐照小鼠血清、9 Gy和12 Gy辐照小鼠血清处理,ATP活力测定法检测细胞活力,流式细胞术检测细胞凋亡情况,RT-qPCR检测Cyp1a1与Cyp1b1 的mRNA表达;将辐照血清处理后的骨髓细胞分为CH223191治疗组与对照组,加入CH223191处理后,比较细胞活力、细胞凋亡、细胞内ROS水平,RT-qPCR检测Cyp1a1与Cyp1b1 的mRNA表达;将骨髓移植小鼠分为CH223191治疗组和对照组,治疗组腹腔注射CH223191,记录小鼠体质量、外周血供体细胞嵌合率及数量、骨髓内供体细胞嵌合率及数量。结果 小鼠经9 Gy或12 Gy辐照后,12 Gy辐照组外周血细胞总数少于9Gy辐照组(P<0.05),而骨髓移植后外周血供体细胞嵌合率无明显差异,并且12 Gy辐照组外周血供体细胞数明显少于9Gy辐照组(P<0.05);小鼠经辐照后,血清内ROS水平上升(P<0.01);经辐照小鼠血清处理的供体骨髓细胞活力降低,凋亡比例升高(均P<0.01),辐照小鼠血清处理的供体骨髓细胞中Cyp1a1与Cyp1b1 mRNA均升高(P<0.05);辐照小鼠血清处理的供体骨髓细胞加入CH223191后,Cyp1a1与Cyp1b1 mRNA水平降低(P<0.05),细胞活力升高,细胞内ROS水平降低,细胞凋亡比例减少(均P<0.01);骨髓移植后,CH223191治疗组小鼠体质量相对于对照组减轻较少,外周血供体细胞嵌合率与细胞数高于对照组,骨髓内供体细胞比例与数量均高于对照组(均P<0.05)。结论 CH223191通过抑制AHR激活,减少辐射诱导的旁观者效应对移植骨髓细胞的损伤,提升小鼠骨髓移植效果。

关键词: 骨髓移植, 辐射诱导的旁观者效应, 芳香烃受体, 活性氧

Abstract: Objective To investigate the effect of radiation-induced bystander effect on donor bone marrow cells during bone marrow transplantation and the protective effect of aryl hydrocarbon receptor (AHR) inhibitor CH223191 on donor bone marrow cells. Methods The mice were divided into 9 Gy irradiated group and 12 Gy irradiated group. The same number of donor bone marrow cells were transplanted after different radiation doses. The total number of peripheral blood cells, chimerism rate and number of peripheral blood donor cells were detected by flow cytometry. Serum reactive oxygen species (ROS) level of unirradiated mice and irradiated mice were detected by ELISA. The donor bone marrow cells were divided into control group, 9 Gy irradiated group and 12 Gy irradiated group and were cultured in vitro. Unirradiated mouse serum, 9 Gy and 12 Gy irradiated mouse serum were added for treatment, respectively. ATP activity assay was used to detect cell vitality, and flow cytometry was used to detect cell apoptosis. The mRNA expression of Cyp1a1 and Cyp1b1 was detected by RT-qPCR. Bone marrow cells treated with irradiated serum were divided into CH223191 treatment group and control group. After adding CH223191 treatment, cell viability, cell apoptosis and intracellular ROS levels were compared, and mRNA expression of Cyp1a1 and Cyp1b1 was detected by RT-qPCR. Bone marrow transplantation mice were divided into CH223191 treatment group and control group. In the treatment group, CH223191 was injected intraperitoneally. The body weight, chimerism rate and number of peripheral blood donor cells, chimerism rate and number of donor cells in bone marrow were recorded. Results The total number of peripheral blood cells in the 12 Gy irradiated group was less than that in the 9 Gy irradiated group (P<0.05), whereas there was no significant difference in the chimeric rate of peripheral blood donor cells after bone marrow transplantation. However, the number of peripheral blood donor cells in the 12 Gy irradiated group was significantly less than that in the 9Gy irradiated group (P<0.05).The serum ROS level increased after irradiation (P<0.01). The activity of donor bone marrow cells was decreased and the percentage of apoptosis was increased (all P<0.01), and the mRNA of Cyp1a1 and Cyp1b1 were increased in irradiated mouse serum (P<0.05). After CH223191 was added into donor bone marrow cells treated with irradiated mouse serum, the mRNA levels of Cyp1a1 and Cyp1b1 were decreased (P<0.05), the cell viability was increased, the intracellular ROS level was decreased, and the percentage of cell apoptosis was decreased (all P<0.01). After bone marrow transplantation, mice in CH223191 treatment group lost less body weight compared with control group, the chimeric rate and cell number of peripheral blood donor cells were higher than control group, and the proportion and number of donor cells in bone marrow were higher than control group (all P<0.05). The total counting of peripheral blood cells in the 12 Gy irradiated group was less than that in the 9 Gy irradiated group (P<0.05), whereas there was no significant difference in the chimeric rates. Conclusions CH223191 inhibits the activation of AHR, reduces the damage of radiation-induced bystander effect on transplanted bone marrow cells, and improves the effect of bone marrow transplantation in mice.

Key words: bone marrow transplantation, radiation-induced bystander effect, aromatic hydrocarbon receptor, reactive oxygen species

中图分类号: