Effect of miR⁃26a⁃3p targeting Survivin on hypoxia/reoxygenation inj ury of H9c2 cardiomyocyte
10.19405/j.cnki.issn1000-1492.2023.11.022
- Author:
Jiancheng Huang
1
;
Hongying Li
1
;
Qingquan Li
2
;
Huijun Zhang
1
;
Xiaobing Li
1
Author Information
1. Dept of Cardiovascular Surgery , The First Hospital of Hebei Medical University , Shijiazhuang 050030The First Hospital of Hebei Medical University , Shijiazhuang 050030
2. Cardiac Intensive Care Unit , The First Hospital of Hebei Medical University , Shijiazhuang 050030
- Publication Type:Journal Article
- Keywords:
miR⁃26a⁃3p;
hypoxia/reoxygenation;
cardiomyocyte;
survivin;
cell apoptosis;
oxidative stress
- From:
Acta Universitatis Medicinalis Anhui
2023;58(11):1934-1941
- CountryChina
- Language:Chinese
-
Abstract:
Objective : To investigate the effects of miR⁃26a⁃3p on rat myocardial cell ( H9c2) injury induced by
hypoxia/reoxygenation (H/R) and its mechanism .
Methods : H9c2 cardiomyocytes in logarithmic growth phase were subjected to hypoxia (1% O2 ) for 6 h , and reoxygenated at different times (2 , 4 , 8 , 12 h) to establish H/R model cell . Normoxia group was also set up , and cell proliferation activity was detected by cell counting kit⁃8 (CCK⁃8) . The level of lactic dehydrogenase (LDH) in cell supernatant was determined by colorimetry . The expression levels of miR⁃26a⁃3p and Survivin mRNA were detected by real⁃time fluorescence quantitative PCR (qRT⁃PCR) . The expression level of Survivin protein in the cells was detected by Western blot . H9c2 cells were transfected with miR⁃26a⁃3p inhibitor and negative control inhibitor NC , Survivin gene siRNA interference plasmid ( si⁃Survivin) and negative control si⁃NC , followed by H/R intervention . CCK⁃8 was used to detect cell proliferation in each group . The activity of superoxide dismutase (SOD) and the content of malonaldehyde (MDA) in cell and the level of LDH in supernatant were determined by colorimetry . The apoptosis level of each group was detected by flow cytometry . The protein expression levels of Bcl⁃2 associated X protein ( Bax) , B ⁃cell lymphoma⁃2 ( Bcl⁃2 ) , cleaved caspase⁃3 and Survivin were detected by Western blot . Targeting relationship between miR⁃26a⁃3p and Survivin gene was determined by dual luciferase .
Results :Compared with the normoxia group , proliferative activity , mRNA and protein expression levels of Survivin in H9c2 cells gradually decreased with the extension of reoxygen ation time (P < 0. 05) , while LDH and expression level of miR⁃26a⁃3p gradually increased ( P < 0. 05) . Downregulating the expression of miR⁃26a⁃3p increased proliferative activity , SOD activity , and expression level of Bcl⁃2 protein in H9c2 cells exposed to H/R ( P < 0. 05) , while MDA content , LDH release amount , apoptosis rate , expression levels of Bax and cleaved caspase⁃3 protein decreased (P < 0. 05) . Survivin deficiency reversed the protective effect of miR⁃26a⁃3p inhibitor on H9c2 cells induced by H/R . Dual luciferase reporter gene assay confirmed that Survivin was the target gene of miR⁃93 ⁃5p .
Conclusion :miR⁃26a⁃3p is highly expressed in cardiomyocyte injury induced by H/R . Inhibition of miR⁃26a⁃3p expression can inhibit H/R⁃induced cardiomyocyte apoptosis and oxidative stress by targeted up⁃regulation of Survivin expression .
- Full text:2024071216295041667miR-26a-3p靶向调...肌细胞缺氧_复氧损伤的影响_黄建成.pdf
