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
计量
- 文章访问数: 3668
- HTML全文浏览量: 1863
- PDF下载量: 28
- 被引次数: 0
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

HTML全文

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）

下载: 全尺寸图片幻灯片

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）

下载: 全尺寸图片幻灯片

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）

下载: 全尺寸图片幻灯片

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）

下载: 全尺寸图片幻灯片

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）

下载: 全尺寸图片幻灯片

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）

下载: 全尺寸图片幻灯片

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

下载: 全尺寸图片幻灯片

参考文献(14)

[1]	Mokhtari-Zaer A,Marefati N,Atkin S L,et al. The protective role of curcumin in myocardial ischemia-reperfusion injury[J]. Journal of Cellular Physiology,2018,234(1):214-222.
[2]	Russo I,Penna C,Musso T,et al. Platelets,diabetes and myocardial ischemia/reperfusion injury[J]. Cardiovascular Diabetology,2017,16(1):71. doi: 10.1186/s12933-017-0550-6
[3]	常国楫,顾金松,孙林. MicroRNA在心肌再灌注损伤应用的研究进展[J]. 昆明医科大学学报,2017,38(5):138-143. doi: 10.3969/j.issn.1003-4706.2017.05.031
[4]	Tony H,Yu K,Qiutang Z. MicroRNA-208a silencing attenuates doxorubicin induced myocyte apoptosis and cardiac dysfunction[J]. Oxidative Medicine and Cellular Longevity,2015,6(2015):597032.
[5]	Guo W,Shi X,Liu A,et al. RNA binding protein QKI inhibits the ischemia/reperfusion-induced apoptosis in neonatal cardiomyocytes[J]. Cell Physiol Biochem,2011,28(4):593-602. doi: 10.1159/000335755
[6]	Kunecki M,Płazak W,Podolec P,et al. Effects of endogenous cardioprotective mechanisms on ischemia-reperfusion injury[J]. Postepy Hig Med Dosw (Online),2017,71(10):20-31. doi: 10.5604/01.3001.0010.3786
[7]	陈耽,李骊华. miR-208a在大鼠心肌缺血再灌注损伤及缺血后处理中的表达及其作用[J]. 重庆医科大学学报,2015,4(40):501-505. doi: 10.13406/j.cnki.cyxb.000494
[8]	Kura B,Szeiffova Bacova B,Kalocayova B,et al. Oxidative Stress-Responsive MicroRNAs in Heart Injury[J]. Int J Mol Sci.,2020,21(1):358. doi: 10.3390/ijms21010358
[9]	Wang F,Yuan Y,Yang P,et al. Extracellular vesicles-mediated transfer of miR-208a/b exaggerate hypoxia/reoxygenation injury in cardiomyocytes by reducing QKI expression[J]. Mol Cell Biochem,2017,431(1-2):187-195. doi: 10.1007/s11010-017-2990-4
[10]	Yan F,Liu R,Zhuang X,et al. Salidroside attenuates doxorubicin-induced cardiac dysfunction partially through activation of QKI/FoxO1 pathway[J]. J Cardiovasc Transl Res,2021,14(2):355-364. doi: 10.1007/s12265-020-10056-x
[11]	Guo W,Jiang T,Lian C,et al. QKI deficiency promotes FoxO1 mediated nitrosative stress and endoplasmic reticulum stress contributing to increased vulnerability to ischemic injury in diabetic heart[J]. J Mol Cell Cardiol,2014,75(10):131-140.
[12]	Al-Salam S,Hashmi S. Myocardial ischemia reperfusion injury:Apoptotic,inflammatory and oxidative stress role of galectin-3[J]. Cell Physiol Biochem,2018,50(3):1123-1139. doi: 10.1159/000494539
[13]	Taki J,Wakabayashi H,Inaki A,et al. 14C-Methionine uptake as a potential marker of inflammatory processes after myocardial ischemia and reperfusion[J]. J Nucl Med,2013,54(3):431-436. doi: 10.2967/jnumed.112.112060
[14]	Li Y,Li Z,Liu J,et al. miR-190-5p alleviates myocardial ischemia-reperfusion injury by targeting PHLPP1[J]. Disease Markers,2021,2021:8709298.

相关文章(20)

[1]	牛志鑫, 汤丽华, 史磊, 洪超, 姚宇峰, 严志凌. MAPK1与NRAS基因多态性与云南汉族人群宫颈上皮内瘤变的相关性, 昆明医科大学学报.
[2]	尹爱桃, 陆永萍, 黄淼, 李祥碧, 尹娇, 徐飞. 初产妇产后早期盆腔脏器脱垂及相关因素分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230412
[3]	董妍, 李雯雯, 崔文龙, 刘杰, 毕卫红. 云南省两县农村老年人慢性病与体力劳动强度相关性分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230303
[4]	梁燕, 王磊, 雷鸣, 陈本超, 孙萍, 李帅, 刘莉, 王倩蓉, 廖曼霖, 马千里. KRAS基因多态性与云南汉族人群非小细胞肺癌的相关性分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230210
[5]	敖健恒, 缪磊, 王慧颖, 章天云, 王涛, 唐永行, 胡利平, 聂胜洁. 手掌接触性生物检材DNA检出率与汗孔密度的相关性研究, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230601
[6]	陆小华, 袁洪新. BTLA、CTLA-4基因多态性与肝癌TACE联合靶向治疗疗效及预后相关性, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230927
[7]	王峰, 杨伟, 钱佃伦, 梅松, 王文杰, 冯科翔, 白向锋. miR-582-5p调控Notch1减轻心肌缺血再灌注损伤的研究, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20221207
[8]	刘蓉, 孙杨, 罗发梦, 陶德智, 张海燕. 炎症性肠病患者疾病接受度与生存质量的相关性, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220507
[9]	马燕粉, 胡健, 蔡德佩, 乔永峰, 王晓琴. 3137例体检人群血清GGT水平和血脂指标的相关性研究, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20221001
[10]	杨佳, 李娅娴, 王莹莹, 肖琳, 李传印, 谭芳, 马千里, 刘舒媛. 云南汉族人群mircoRNA-149、mircoRNA-219、mircoRNA-let-7基因多态性与非小细胞肺癌发生和发展的相关性, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20211037
[11]	崔朴梅, 陈太方, 陈超, 梁永梅, 徐悦. 云南某医院2016年至2019年抗菌药物使用与常见细菌耐药性的相关性分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20211127
[12]	吴欣, 李云, 邹品飞, 李梅玲, 普勇斌. 肌骨超声检查在风湿性膝关节炎患者中检出率及与膝关节功能的相关性, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210814
[13]	黄瑛, 李翔, 江姗, 刘冰, 张燕玲, 李杨, 何花, 武彦, 罗云, 徐俊. 2019-nCoV感染者胸部影像学与实验室指标相关性分析, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20211232
[14]	杨媛, 梁红敏, 朱桂敏, 杨寒凝, 曹维涵, 陆永萍. 携IL-6单克隆抗体靶向微泡破坏技术在兔MI/RI损伤中的应用, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210222
[15]	王娟娟, 陈雨青, 邓茜, 高健. 心肌酶谱与儿童1型糖尿病酮症酸中毒严重程度的相关性, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210129
[16]	徐剑. CyPA与慢性肾衰竭相关性及作用研究, 昆明医科大学学报.
[17]	梅雪. 年龄、性别因素对高原地区高尿酸血症的相关性分析, 昆明医科大学学报.
[18]	郑红梅. 65例皮肌炎患者血清肌酶变化与年龄、性别相关性分析, 昆明医科大学学报.
[19]	赵金友. 云南省城市空巢老人孤独状况与生命质量相关性分析, 昆明医科大学学报.
[20]	张红. 外周血红细胞和血红蛋白因素与高原地区高尿酸血症患者相关性分析, 昆明医科大学学报.