Period 2 protein inhibits cardiomyocyte death induced by β<sub>1</sub>-adrenoceptor autoantibodies

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Period 2 protein inhibits cardiomyocyte death induced by β₁-adrenoceptor autoantibodies

Basic Medicine | 更新时间：2026-02-13

- Period 2 protein inhibits cardiomyocyte death induced by β₁-adrenoceptor autoantibodies
- Journal of Army Medical University Vol. 44, Issue 9, Pages: 898-905(2022)
- 作者机构：
  
  1.030001 太原，山西医科大学基础医学院：基础医学研究中心
  2.030001 太原，山西医科大学基础医学院：形态学实验室
  3.030012 太原，山西医科大学附属人民医院
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(31871177);the Post-graduate Education Innovation Project of Shanxi Province(2021Y409)
- DOI：10.16016/j.2097-0927.202110010
  CLC： R322.11;R329.25;R341
- Received：07 October 2021，
  
  Revised：2021-12-08，
  
  Published：15 May 2022
- 稿件说明：
移动端阅览
Pengjia LI, Weiwei JIA, Jiebei LU, et al. Period 2 protein inhibits cardiomyocyte death induced by β₁-adrenoceptor autoantibodies[J]. Journal of Army Medical University, 2022, 44(9): 898-905.
DOI：

Pengjia LI, Weiwei JIA, Jiebei LU, et al. Period 2 protein inhibits cardiomyocyte death induced by β₁-adrenoceptor autoantibodies[J]. Journal of Army Medical University, 2022, 44(9): 898-905. DOI： 10.16016/j.2097-0927.202110010.

摘要

目的

2

探讨周期蛋白2(period 2，Per2)在β

1

-肾上腺素受体(β

1

-adrenergic receptor，β

1

-AR)自身抗体(β

1

-adrenergic receptor autoantibodies，β

1

-AA)诱导的心肌细胞死亡中的作用。

方法

2

选取6~8周龄SD雄性大鼠16只随机分为主动免疫组[模型组，采用β

1

-肾

上腺素受体细胞外第二环抗原肽段(β

1

-AR-EC Ⅱ)对大鼠进行主动免疫

]

和对照组(同期注射Na

2

CO

3

等溶液)，每组8只。8周时取大鼠血清，提纯β

1

-AA。购买H9c2心肌细胞，随机分为Control组、β

1

-AA组和β

1

-AR+β

1

-AA组，CCK-8法检测细胞存活率(

n

=8)；Control组和β

1

-AA(1 μmol/L)组细胞经地塞米松同步化4 h后，Western blot检测两组不同时间点心肌细胞Per2的表达，JTK_CYCLE分析昼夜节律参数(

n

=11)；分别采用慢病毒shPer2和Per2敲低或过表达心肌细胞中Per2的表达，RT-PCR和Western blot检测Per2的表达(

n

=6)；在慢病毒敲低Per2(

n

=8)和过表达Per2(

n

=10)基础上，结合β

1

-AA处理，CCK-8法检测心肌细胞存活率。

结果

2

CCK-8结果显示，β

1

-AA诱导H9c2细胞存活率明显降低(

P

<

0.05)。Western blot结果显示β

1

-AA显著抑制心肌细胞Per2蛋白的节律性表达(JTK_CYCLE

P

>

0.05)，其中CT8和CT16降低最为明显(

P

<

0.01)。慢病毒敲低Per2表达可引起心肌细胞存活率降低；在此基础上进行β

1

-AA处理，细胞存活率进一步降低(

P

<

0.001)。而过表达Per2可明显逆转β

1

-AA诱导的心肌细胞存活率下降(

P

<

0.001)。

结论

2

Per2抑制β

1

-AA诱导的H9c2心肌细胞死亡。

Abstract

Objective

2

To investigate the role of period 2 (Per2) protein in the death of cardiomyocytes induced by β

1

-adrenergic receptor autoantibodies (β

1

-AA).

Methods

2

Sixteen male SD rats aged 6~8 weeks were randomly divided into active immunization (model) group and control group (

n

=8). The model group was immunized with the second extracellular loop of beta1-adrenoceptor (β

1

-AR-EC Ⅱ)

and the control group was injected with Na

2

CO

3

and other solutions. The rat serum was subsequently collected at 8 weeks

followed by the purification of β

1

-AA. H9c2 cardiomyocyte were selected and randomly divided into control group

β

1

-AA group

and β

1

-AR+β

1

-AA group. Cell viability of each group was detected by CCK-8 assay (

n

=8). Then H9c2 cells in th

e control group and β

1

-AA 1 μmol/L group were synchronized with dexamethasone for 4 h

the expression of Per2 in cardiomyocytes at different circadian time (CT) points was measured by Western blotting

and JTK_CYCLE was used to analyze the circadian rhythm parameters (

n

=11). Moreover

Per2 in H9c2 cells was knocked down or overexpressed by lentiviral shPer2 and lentiviral Per2

respectively; RT-PCR and Western blotting were performed to detect the changes of Per2 expression (

n

=6). On the basis of knockdown (

n

=8) or overexpression (

n

=10) of Per2

the H9c2 cells were further treated with β

1

-AA

and the cell survival rate was tested by CCK-8 assay.

Results

2

CCK-8 assay showed that the survival rate of H9c2 cells was significantly decreased after β

1

-AA treatment (

P

<

0.05). Western blotting demonstrated that β

1

-AA remarkably inhibited the rhythmic expression of Per2 protein in the cardiomyocytes (JTK_CYCLE

P

>

0.05)

with the decrease at CT8 and CT16 most obviously (

P

<

0.01). Knockdown of Per2 expression reduced the survival rate of cardiomyocytes

which was further lowered after β

1

-AA treatment (

P

<

0.001). However

overexpression of Per2 notably reversed the decline in H9c2 survival rate induced by β

1

-AA (

P

<

0.001).

Conclusion

2

Per2 protein inhibits β

1

-AA induced H9c2 cardiomyocyte death.

关键词

Keywords

references

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