MiR-208b-3p aggravates energy metabolism disorders in mice with heart failure by inhibiting mitochondrial gene expression

Shuangshan ZHOU; Yuan LIU; Ping YIN; Yujun SHI; Li SU

doi:10.16016/j.2097-0927.202402085

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MiR-208b-3p aggravates energy metabolism disorders in mice with heart failure by inhibiting mitochondrial gene expression

Basic Medicine | 更新时间：2026-01-09

- MiR-208b-3p aggravates energy metabolism disorders in mice with heart failure by inhibiting mitochondrial gene expression
- Journal of Army Medical University Vol. 46, Issue 16, Pages: 1857-1866(2024)
- 作者机构：
  
  1.400014 重庆，重庆医科大学附属第二医院心血管内科
  2.610041 成都，四川大学华西医院病理研究室
- 作者简介：
- 基金信息：
  
  the General Program of Natural Science Foundation of Chongqing(cstc2021jcyj-msxmX0208)
- DOI：10.16016/j.2097-0927.202402085
  CLC： R341;R394.1;R541.602
- Received：27 February 2024，
  
  Revised：2024-04-14，
  
  Published：30 August 2024
- 稿件说明：
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Shuangshan ZHOU, Yuan LIU, Ping YIN, et al. MiR-208b-3p aggravates energy metabolism disorders in mice with heart failure by inhibiting mitochondrial gene expression[J]. Journal of Army Medical University, 2024, 46(16): 1857-1866.
DOI：

Shuangshan ZHOU, Yuan LIU, Ping YIN, et al. MiR-208b-3p aggravates energy metabolism disorders in mice with heart failure by inhibiting mitochondrial gene expression[J]. Journal of Army Medical University, 2024, 46(16): 1857-1866. DOI： 10.16016/j.2097-0927.202402085.

摘要

目的

探究miR-208b-3p对主动脉弓缩窄(transverse aortic constriction

TAC)诱导的心力衰竭小鼠能量代谢的作用及机制。

方法

将24只小鼠按照随机数字表法分为假手术组(Sham组

=6)和手术组(

=18)。手术组采用TAC建立心力衰竭模型; 假手术组采用TAC相同手术方式

但不结扎主动脉弓。于TAC术后第2周再随机分为Antagomir组(

=6)、Antagomir-NC组(

=6)及TAC组(

=6)

Antagomir组、Antagomir-NC组尾静脉分别注射miR-208b-3p antagomir (800 μg)及miR-208b-3p antagomir阴性对照(800 μg)试剂

每周2次

连续4周。术后第6周行超声心动图评估各组小鼠心功能。HE染色及天狼星红染色观察小鼠心肌组织病理学形态。ATP检测试剂盒检测各组小鼠心肌组织ATP水平。RT-qPCR检测小鼠心肌组织miR-208b-3p、线粒体基因(ND1、ND2、ND3、ND4、ND4L、ND5、ND6、CO1、CO2、CO3、CYTB、ATP6、ATP8)及POLRMT、12SrRNA的表达。双荧光素酶报告基因实验检测miR-208b-3p与潜在靶基因POLRMT的相互结合作用。Western blot检测心肌组织POLRMT及ND1、CO2、CYTB、ATP8蛋白表达水平。

结果

与Sham组相比

TAC组小鼠心肌miR-208b-

3p表达显著增加(

0.05)。超声检测结果显示

Antagomir组小鼠心脏的射血分数、收缩与舒张功能较TAC组明显改善(

0.05)。组织切片染色提示

Antagomir组小鼠的心肌细胞肥大、排列紊乱、心肌间质炎细胞浸润等有明显改善(

0.05)。与TAC组相比

Antagomir组ATP水平明显升高(

0.05);POLRMT及线粒体基因转录产物(12SrRNA和13个线粒体基因编码多肽)表达量均显著升高(

0.05)

但SDHA、SDHB无变化。双荧光素酶报告基因实验显示miR-208b-3p与POLRMT的CDS区有结合作用。Antagomir组小鼠心肌组织POLRMT、ND1、CO2、CYTB、ATP8蛋白水平表达较TAC组显著增加(

0.05)。

结论

miR-208b-3p可通过靶向POLRMT抑制线粒体基因的表达

加剧线粒体能量代谢障碍

并加重心力衰竭小鼠心功能不全及心室重塑。

Abstract

Objective

To explore the effect and mechanism of miR-208b-3p on energy metabolism in mice with heart failure (HF) induced by transverse aortic constriction (TAC).

Methods

Twenty-four mice were randomly divided into sham operation group (Sham group

=6) and surgery group (

=18).TAC was used to establish an HF model in the surgical group

the sham group received the same surgical procedures as TAC

but no ligation of the transverse aortic arc.At the second week after TAC

the surgery group was randomly divided into Antagomir group (

=6)

Antagomir-NC group (

=6) and TAC group (

=6).The mice of the Antagomir group and the Antagomir-NC group were injected with miR-208b-3p antagomir reagent (800 μg) and miR-208b-3p antagomir negative control reagent (800 μg)

respectively by tail vein

twice a week

for 4 consecutive weeks.Echocardiography was performed at the 6th week after surgery to evaluate the cardiac function.HE staining and Sirius red staining were used to observe myocardial histopathology in mice.ATP assay was employed to detect the ATP level in myocardial tissues.RT-qPCR was applied to detect the expression of miR-208b-3p

mitochondrial genes (ND1

ND2

ND3

ND4

ND4L

ND5

ND6

CO1

CO2

CO3

CYTB

ATP6 and ATP8)

POLRMT and 12S rRNA in myocardial tissues.Double luciferase reporter assay was conducted to detect the interaction between miR-208b-3p and the potential target gene POLRMT.Western blotting was utilized to detect the changes in the protein levels of POLRMT and ND1

CO2

CYTB and ATP8 in myocardial tissues.

Results

The expression of miR-208b-3p was significantly higher in myocardial tissues of the TAC group than the Sham group (

0.05).Echocardiography revealed that the ejection fraction

systolic and diastolic functions were significantly improved in the Antagomir group than the TAC group (

0.05).Pathological observation showed significantly improved cardiomyocyte hypertrophy

arrangement disorder and myocardial interstititial cell infiltration in the Antagomir group (

0.05).Compared with the TAC group

the ATP level was significantly increased (

0.05)

the expression levels of POLRMT and mitochondrial gene transcripts (12SrRNA and 13 mitochondrial gene-coding polypeptides) were significantly increased (

0.05)

but there were no changes in SDHA and SDHB levels in the Antagomir group.Double luciferase reporter assay indicated that miR-208b-3p bound to the CDS region of POLRMT.The protein levels of POLRMT

ND1

CO2

CYTB and ATP8 were significantly increased in the myocardial tissues in the Antagomir group than the TAC group (

0.05).

Conclusion

MiR-208b-3p inhibits the expression of mitochondrial genes by targeting POLRMT

aggravates mitochondrial energy metabolism disorder

and deteriorates cardiac insufficiency and ventricular remodeling in HF mice.

关键词

Keywords

references

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