Effect of Picroside Ⅱ on Expression of microRNA-1 in H<sub>2</sub>O<sub>2</sub>-induced H9c2 Cardiomyocytes Damage

Qing-rui SHAO; Wei-de LING; Jian-zhe LI

doi:10.13422/j.cnki.syfjx.20190802

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Effect of Picroside Ⅱ on Expression of microRNA-1 in H₂O₂-induced H9c2 Cardiomyocytes Damage

Pharmacology | 更新时间：2021-02-09

- Effect of Picroside Ⅱ on Expression of microRNA-1 in H₂O₂-induced H9c2 Cardiomyocytes Damage
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 8, Pages: 77-82(2019)
- 作者机构：
  
  广西中医药大学　附属瑞康医院，南宁　 530011
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190802
  CLC： R2-0;R22;R285.5
- Received：10 October 2018，
  
  Published Online：04 January 2019，
  
  Published：20 April 2019
- 稿件说明：
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Qing-rui SHAO, Wei-de LING, Jian-zhe LI. Effect of Picroside Ⅱ on Expression of microRNA-1 in H₂O₂-induced H9c2 Cardiomyocytes Damage[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 77-82.
DOI：

Qing-rui SHAO, Wei-de LING, Jian-zhe LI. Effect of Picroside Ⅱ on Expression of microRNA-1 in H₂O₂-induced H9c2 Cardiomyocytes Damage[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 77-82. DOI： 10.13422/j.cnki.syfjx.20190802.

摘要

目的：

研究胡黄连苦苷Ⅱ(picroside Ⅱ)对过氧化氢(H

)诱导的大鼠H9c2心肌细胞凋亡中微小RNA-1 (microRNA-1

miR-1)及下游靶基因抗凋亡因子B淋巴细胞瘤-2(B-cell lymphoma-2

Bcl-2)表达的影响，探讨胡黄连苦苷Ⅱ的心肌保护作用及其机制。

方法：

将H9c2心肌细胞随机分为正常组，模型组(H

，200 μmol·L

-1

)，胡黄连苦苷Ⅱ (50

100

200 μmol·L

-1

)+H

(200 μmol·L

-1

)组，胡黄连苦苷Ⅱ 200 μmol·L

-1

组。胡黄连苦苷Ⅱ与心肌细胞孵育6 h后，加入H

继续培养2 h。药物处理结束后，噻唑蓝(methyl thiazolyl tetrazolium

MTT)比色法检测细胞存活率，比色法测定细胞培养液中乳酸脱氢酶(lactate dehydrogenase

LDH)活性，4′，6-二脒基-2-苯基吲哚(destination access point identifier

DAPI)染色和细胞凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3 (cysteinyl aspartate specific proteinase-3，Caspase-3)检测评估细胞凋亡，实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞中Caspase-3，Bcl-2，miR-1 mRNA的表达，蛋白免疫印迹法(Western blot)检测细胞抗凋亡因子Bcl-2蛋白的表达。

结果：

与正常组相比，H

能显著降低细胞生存率，增加细胞凋亡率，且Caspase-3活性和mRNA表达均增加(

0.01)，同时伴有miR-1 mRNA表达水平的上调及其下游靶基因抗凋亡因子Bcl-2 mRNA表达的下调(

0.01)。与模型组比较，胡黄连苦苷Ⅱ预处理可明显升高细胞存活率，明显降低LDH活性，降低细胞凋亡率，明显降低Caspase-3活性和mRNA表达(

0.05，

0.01)，并使Bcl-2 mRNA表达升高(

0.05，

0.01)，Western blot结果表明，胡黄连苦苷Ⅱ能明显下调其下游靶基因抗凋亡因子Bcl-2蛋白表达(

0.05，

0.01)。

结论：

胡黄连苦苷Ⅱ对H

诱导的H9c2心肌细胞凋亡具有保护作用，其机制与下调 miR-1的表达进而上调其靶基因Bcl-2的表达有关。

Abstract

Objective:

To study the effect of picroside Ⅱ on the expression of microRNA-1 (miR-1) in the H

-induced H9c2 cardiomyocytes damage

in order to explore the mechanism of picroside Ⅱ in protecting H9c2 cardiomyocytes from oxidative stress.

Method:

H9c2 cardiomyocytes were divided into 6 groups: control group

model group (H

200 μmol·L

-1

)

picroside Ⅱ (50

100

200 μmol·L

-1

)+ H

(200 μmol·L

-1

) group and picroside Ⅱ (200 μmol·L

-1

) group. Picroside Ⅱ group was incubated with picroside Ⅱ for 6 h and then cultured with H

for 2 h. At the end of drugs treatment

the cell viability and the cellular damage of cardiomyocytes were respectively assessed by 3-(4

5-dimethylthiazol-2-yl)-2

5-diphenyl tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. 4′

6-diamidino-2-phenylindole (DAPI) staining and cysteinyl aspartate specific proteinase-3 (Caspase-3) test were used to evaluate cell apoptosis. The mRNA expressions of Caspase-3

B-cell lymphoma-2 (Bcl-2) and miR-1 were measured by Real-time polymerase chain reaction (Real-time PCR). The protein expression of Bcl-2 was detected by Western blot.

Result:

Compared with the control group

could significantly decrease the cell viability and increase the rate of apoptosis

up-regulate mRNA expression of Caspase-3 and miR-1

and down-regulate expression of Bcl-2 in H9c2 cells (

0.01). Compared with the model group

the cell survival rate was significantly increased by pretreating with Picroside Ⅱ for 6 h (

0.05

0.01). Picroside Ⅱ not only decreased the rate of apoptosis and up-regulated mRNA expression of Bcl-2

but also down-regulated mRNA expressions of Caspase-3 and miR-1 (

0.05

0.01). Western blot showed that picroside Ⅱ can significantly enhance the expression of Bcl-2 (

0.05

0.01).

Conclusion:

Picroside Ⅱ has a protective effect on H9c2 cells from H

-induced cardiomyocyte injury by down-regulating mRNA-1 expression and up-regulating the expression of the downstream Bcl-2.

关键词

Keywords

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Effect of Picroside Ⅱ on Expression of microRNA-1 in H2O2-induced H9c2 Cardiomyocytes Damage

DOI：10.13422/j.cnki.syfjx.20190802

摘要

Abstract

关键词

Keywords

references

Effect of Picroside Ⅱ on Expression of microRNA-1 in H₂O₂-induced H9c2 Cardiomyocytes Damage