miR-208a通过调控QKI5表达对大鼠心肌缺血再灌注损伤的影响

杨伟; 陈洪艳; 陈文栋; 王燕琼; 白向锋

doi:10.12259/j.issn.2095-610X.S20220724

miR-208a通过调控QKI5表达对大鼠心肌缺血再灌注损伤的影响

doi: 10.12259/j.issn.2095-610X.S20220724

1.
昆明医科大学第一附属医院麻醉科
2.
超声科
3.
心脏大血管外科，云南昆明　650032

基金项目: 云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目[2019FE001（-045）]；云南省教育厅科学研究基金资助项目[2021J0230]

详细信息

作者简介:
杨伟（1980～），男，河南信阳人，医学博士，副主任医师，主要从事心血管手术麻醉工作

通讯作者:
白向锋，E-mail：49976790@qq.com

中图分类号: R614；R541
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出版历程
- 收稿日期: 2022-04-16
- 网络出版日期: 2022-06-25
- 刊出日期: 2022-07-14

Effects of miR-208a on Myocardial Ischemia-reperfusion Injury by Regulating the Expression of QKI5 in Rats

1.
Dept. of Anesthesiology
2.
Dept. of Ultrasound
3.
Dept. of Cardiovascular Surgery，The 1st Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032，China

摘要

摘要: 目的研究miR-208a调控QKI5表达对大鼠心肌缺血再灌注损伤的影响及作用机制。方法将雄性SD大鼠32只随机选择分为假手术组（Sham组）、缺血再灌注组（I/R组）、I/R+antagomir阴性对照组（I/R+anta-miR-NC组）和I/R+miR-208a antagomir 组（I/R+anta-miR-208a组），构建大鼠心肌I/R模型后检测各组大鼠心脏左室射血分数（LVEF）、左室短轴缩短率（LVFS）和左室每搏量（SV）；ELISA法检测血清肌酐激酶同工酶（CK-MB）和心肌肌钙蛋白（cTnI）以及IL-1β、IL-6和肿瘤坏死因子（TNF-α）水平；采用HE染色观察各组心肌组织病理变化，采用TUNEL染色检测心肌组织细胞凋亡；RT-PCR检测心肌组织miR-208a和QKI5 mRNA表达，Western blot检测心肌组织QKI5蛋白和凋亡蛋白Caspase-3的相对表达水平。结果与Sham组比较，I/R组中LVEF、LVFS、SV值降低，心肌细胞凋亡率升高，血清中CK-MB、cTnI和IL-1β、IL-6、TNF-α水平及心肌组织中miR-208a和Caspase-3表达升高，而QKI5表达水平降低（P < 0.05）；与I/R+anta-miR-NC组比较，I/R+anta-miR-208a组中LVEF、LVFS、SV值升高，心肌细胞凋亡率降低，血清中CK-MB、cTnI和IL-1β、IL-6、TNF-α及心肌组织中miR-208a和Caspase-3表达水平降低，而QKI5表达升高（P < 0.05）。相关性分析发现miR-208a与QKI5表达呈负相关。结论抑制miR-208a可能通过调控QKI5表达减轻心肌缺血再灌注损伤中炎症反应，发挥心肌保护作用。
- 心肌缺血再灌注损伤 /
- 微小RNA-208a /
- QKI5蛋白 /
- 相关性
Abstract: Objective To investigate the effect and mechanism of miR-208a on myocardial injury induced by myocardial ischemia reperfusion by regulating the expression of QKI5 in rats. Methods Thirty-two male SD rats were randomly divided into sham group, I/R group, I/R+anta-miR-NC group and I/R+anta-miR-208a group. The rat model of myocardial I/R was established, and the LVEF, LVFS and SV were detected in each group. The levels of CK-MB, cTnI, IL-1β, IL-6, TNF-αwere measured by ELISA. HE staining was used to observe the pathological changes of myocardial tissue in each group, and TUNEL staining was used to detect myocardial cell apoptosis.RT-PCR was used to detect the expression of miR-208a and QKI5 mRNA in myocardial tissue, and Western blot was used to detect the realative expression levels of QKI5 protein and Caspase-3 in myocardial tissue. Results Compared with the Sham group, the levels of LVEF, LVFS, SV in the I/R group were decreased, while the apoptosis rate of cardiomyocytes was increased, and the levels of CK-MB, cTnI, IL-1β, IL-6, TNF-α in serum and the expressions of miR-208a and Caspase-3 in the myocardial tissue were increased, and the expressions of QKI5 were decreased （P < 0.05）. Compared with the I/R+anta-miR-NC group, the levels of LVEF, LVFS, SV in the I/R+anta-miR-208a group were increased, while the apoptosis rate of cardiomyocytes was decreased, and the levels of CK-MB, cTnI, IL-1β, IL-6, TNF-α in serum and the expressions of miR-208a and Caspase-3 in the myocardial tissue were decreased, and the expressions of QKI5 were increased （P < 0.05）. Correlation analysis found that the expression of miR-208a and QKI5 had negative correlation in the I/R rats. Conclusion Inhibition of miR-208a may alleviate the inflammatory response to exert myocardial protective function in myocardial ischemia-reperfusion injury by regulating the expression of QKI5.
- Myocardial ischemia-reperfusion injury /
- miR-208a /
- QKI5 protein /
- Correlation

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图 1 miR-208a对大鼠心脏功能的影响

A：LVEF值；B：FS值；C：SV值。与Sham组相比，^*P < 0.05；与I/R组相比，^#P < 0.05；与anta-miR-NC组相比，^▲P < 0.05。

Figure 1. Effect of anta-miR-208a on cardiac function in rats（$\bar x \pm s $，n = 8）

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图 2 各组心肌组织病理变化（HE ×200）

A：Sham组；B：I/R组；C：I/R+anta-miR-NC组；D：I/R+anta-miR-208a组。

Figure 2. Pathological changes of myocardial tissue in each group （HE ×200）

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图 3 miR-208a对I/R大鼠心肌损伤标注物和炎性因子的影响

A、B：大鼠血清心肌损伤标志物CK-MB和cTnI水平；C、D、E：大鼠血清炎性因子IL-1β、IL-6和TNF-α水平。与Sham组相比，^*P < 0.05；与I/R组相比，^#P < 0.05；与anta-miR-NC组相比，^▲P < 0.05。

Figure 3. Effects of miR-208a expression on myocardial injury makers and inflammatory factors in I/R rats（$\bar x \pm s $，n = 8）

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图 4 各组心肌组织细胞凋亡情况（TUNEL×200）

A：Sham组；B：I/R组；C：I/R+anta-miR-NC组；D：I/R+anta-miR-208a组；E：大鼠心肌细胞凋亡率。与Sham组相比，^*P < 0.05；与I/R组相比，^#P < 0.05；与anta-miR-NC组相比，^▲P < 0.05。

Figure 4. Apoptosis of myocardial tissue cells in each group （TUNEL×200）（$\bar x \pm s $，n = 8）

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图 5 各组大鼠心肌组织miR-208a和QKI5 mRNA相对表达水平

与Sham组相比，^*P < 0.05；与I/R组相比，^#P < 0.05；与anta-miR-NC组相比，^▲P < 0.05。

Figure 5. Relative expression levels of miR-208a and QKI5 mRNAin myocardial tissue of rats in each group （$\bar x \pm s $，n = 8）

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图 6 各组大鼠心肌组织中蛋白的相对表达

A：心肌组织中QKI5和Caspase3的WB图；B：心肌组织中QKI5和Caspase3相对表达量。与Sham组相比，^*P < 0.05；与I/R组相比，^#P < 0.05；与anta-miR-NC组相比，^▲P < 0.05。

Figure 6. Relative expression levels of protein in myocardial tissue of rats in each group（$\bar x \pm s $，n = 8）

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图 7 miR-208a和QKI5的相关性

A：数据库预测miR-208a与QKI5结合区域；B：miR-208a与QKI5表达的相关性。

Figure 7. Correlation between miR-208a and QKI5

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