Hydroxyurea ameliorates mouse hypoxia through inducing embryonic hemoglobin

Xiaoying ZHOU; Li JIAO; Yu CUI; Xiaoxu LI; Yidong YANG; Chengzhong YANG; Chenyuan WANG; Jian HUANG

doi:10.16016/j.2097-0927.202206100

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Hydroxyurea ameliorates mouse hypoxia through inducing embryonic hemoglobin

Military Medicine | 更新时间：2026-01-09

- Hydroxyurea ameliorates mouse hypoxia through inducing embryonic hemoglobin
- Journal of Army Medical University Vol. 45, Issue 2, Pages: 93-101(2023)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)高原军事医学系高原生理学与病理学教研室
- 作者简介：
- 基金信息：
  
  the Key Special Program of "Thirteenth Five-year Plan" of Logistic Scientific Research of PLA(BWS17J031);the Youth Training Plan of Military Medical Science and Technology(20QNPY002)
- DOI：10.16016/j.2097-0927.202206100
  CLC： R-332;R594.3;R973.3
- Received：17 June 2022，
  
  Revised：2022-08-25，
  
  Published：30 January 2023
- 稿件说明：
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Xiaoying ZHOU, Li JIAO, Yu CUI, et al. Hydroxyurea ameliorates mouse hypoxia through inducing embryonic hemoglobin[J]. Journal of Army Medical University, 2023, 45(2): 93-101.
DOI：

Xiaoying ZHOU, Li JIAO, Yu CUI, et al. Hydroxyurea ameliorates mouse hypoxia through inducing embryonic hemoglobin[J]. Journal of Army Medical University, 2023, 45(2): 93-101. DOI： 10.16016/j.2097-0927.202206100.

摘要

目的

探讨羟基脲(hydroxyurea

HU)对密闭缺氧小鼠的保护作用及机制。

方法

将60只6~8周龄、体质量18~22 g的雄性C57BL/6J小鼠随机分成常氧对照组(NC组

=10)、常氧HU组(NHU组

=10)、缺氧对照组(HC组

=20)、缺氧HU组(HHU组

=20)。HU组每天腹腔注射HU 100 mg/kg; 对照组每天注射等量生理盐水。4周后取NHU组和NC组小鼠的骨髓进行转录组测序; 再将HHU组和HC组置于密封瓶

进行密闭缺氧

记录其生存时间; 在缺氧15 min时

取脑、肾组织进行免疫组化

同时取未缺氧的NHU组和NC组脑、肾组织进行对照; 并对骨髓进行RT-qPCR验证

外周血进行Western blot验证。

结果

HHU组小鼠密闭缺氧的生存时间显著延长(

0.05);脑和肾脏缺氧程度显著降低(

0.05)。外周血氧亲和力显著提高

且P50与小鼠生存时间存在显著的负相关关系(Pearson=-0.738

0.01)。转录组共检测到65 389个基因

其中NHU组上调基因50个

下调基因272个(log2 FC≥1

0.05)

对差异显著的基因进行功能富集

结果发现在分子组成上

血红蛋白复合体居于首位; 分子功能上

主要富集到影响血红蛋白各亚基结合的分子; 生物学功能上主要富集到红细胞发育相关的生物学过程。差异分子中

Erfe

、

Hbb

、

Hbb

1、

Hba

1.5倍以上

而下调基因有红系发育相关基因

Klf

1、

Sox

6、

Gata

1等; 经验证

NHU组

Hbb

、

Hbb

1、

Hba

的mRNA显著升高

外周血HbE (

-globin)蛋白表达增加

差异具有统计学意义(

0.05)。

结论

HU对密闭缺氧小鼠具有明显的保护作用

可能与HU诱导高氧亲和力的胚胎样血红蛋白表达

从而促进携氧的作用有关。

Abstract

Objective

To investigate the ameliorative effect and mechanism of hydroxyurea (HU) on hypoxia mice in a closed environment.

Methods

Sixty male C57BL/6J mice (6~8 weeks old

weighing 18~22 g) were randomly divided into normoxia control group (NC

=10)

normoxia HU group (NHU

=10)

hypoxia control group (HC

=20)

and hypoxia HU group (HHU

=20).The HU mice were injected intraper

itoneally with 100 mg/kg HU daily for 4 weeks

and the control mice were injected with same volume saline for 4 weeks.Then bone marrow was extracted for transcriptome sequence in the NC and NHU groups.The mice in the HC and HHU groups were settled in a sealed bottle to observe their survival

and the brains and kidneys were isolated for immunohistochemistry when these mice were in hypoxia for 15 min

and those from the NHU and NC groups served as control.RT-qPCR and Western blotting were adopted to verify the mRNA and protein expression of bone marrow and peripheral blood molecules

respectively.

Results

The survival time was significantly longer in the HHU group (

0.05)

and the hypoxia of the brains and kidneys were ameliorated (

0.05).The oxygen affinity of peripheral blood was obviously raised

and partial pressure of oxygen 50(P50) was negatively correlated with survival time (Pearson=-0.738

0.01).A total of 65 389 genes were found in transcription sequencing

including 50 up-regulated and 272 down-regulated genes in the mice from the NHU group (log2 FC≥1

0.05).Gene ontology (GO) enrichment analysis indicated that hemoglobin subunits (GO cellular component)

hemoglobin complex (GO cellular function) and erythrocyte development (GO biological process) were significant enriched.Among the differentially expressed genes

Erfe

Hbb

1 and

Hba

were up to 1.5 fold

and the down-regulated genes were erythropoiesis-related genes

Klf

Sox

Gata

etc

.Our results indicated that the mRNA levels of

Hbb

1 and

Hba

were notably enhanced in the bone marrow of the NHU group

and the protein leve

l of HbE (

-globin) protein was remarkably increased in the peripheral blood (

0.05).

Conclusion

HU exerts protective effect on the mice in a closed hypoxic condition

which may be related with its inducing embryonic hemoglobin with high oxygen affinity

and thus promoting oxygen carrying capacity.

关键词

Keywords

references

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