Protective effect and its related with autophagy of growth differentiation factor 15 over expression on myocardial ischemia/reperfusion
10.13699/j.cnki.1001-6821.2017.10.012
- VernacularTitle:生长分化因子15过表达对心肌缺血再灌注的保护作用及与其自噬的关系
- Author:
Lei CAO
1
;
Qian GUO
;
Zhang-Zhao WANG
;
Xin-Ran ZHANG
;
Jian-Jie JIAO
;
Jing-Hua HE
;
Zhao-Yan QIANG
Author Information
1. 天津医科大学基础医学院药理学系
- Keywords:
growth differentiation factor 15;
lentivirus infection;
cardiomyocyte;
hypoxia/reoxygenation
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(10):909-912
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of growth differentiation factor 15 (GDF15) on H9c2 cardiomyocytes against ischemia/reperfusion injury(I/R) by constructing GDF15 stable over expression of myocardial cell lines.Methods To establish the model of hypoxia/reoxygenation,transfect targeted GDF15 over expression lentivirus and negative empty vector lentivirus into H9c2 cells,and select the H9c2 cells of GDF15 gene over expression stably.Randomly assigned to control group,transfected with empty vector(H9c2/Sham) and GDF15 over expression group (H9c2/GDF15).Cell viability was detected by MTT.And lactate dehydrogenase (LDH) in supernatant of cell culture was measured by LDH kit.The expression levels of apoptosis-related protein Caspase-3,Bcl-2,Bax and autophagy-related protein Beclin-1,LC3-B protein were observed with Western-blotting.Results Compared with control group (60.47 ± 5.03) ％ and H9c2/Sham group (63.21 ± 17.13) ％,the H9c2/GDF15 group (74.93 ± 10.03) ％ increased significantly,the difference was statistically significant after I/R 2 h (P ＜ 0.01).Compared with control group 108.72 ± 7.20,the activity of LDH in H9c2/GDF15 group 50.29 ± 10.05,the difference was statistically significant (P＜0.01).To apoptosis-related protein Caspase-3,the expression in H9c2/GDF15 group (gray value 67.68 ±0.47) was lower than control group (gray value 99.73 ±0.61),the difference was statistically significant after I/R2 h(P ＜0.01).The Bcl-2/Bax in H9c2/GDF15 group 82.52 ± 1.39 was higher than control group 72.52 ±2.05 and H9c2/Sham group 39.55 ± 0.46,the difference was statistically significant after I/R 2 h (P＜0.01).To autophagy-related protein Beclin-1,the expression in H9c2/GDF15 group (gray value 88.39 ±0.72) was higher than control group (gray value 58.93 ± 0.64) and H9c2/Sham group (gray value 36.71 ±0.35),the difference was statistically significant(P ＜0.01).The LC3-Ⅱ/LC3-Ⅰ in H9c2/GDF15 group (3.32 ±0.04) was higher than control group(2.11 ± 0.02),the difference was statistically significant (P ＜0.01).Conclusion Up-regulation of GDF15 in H9c2 cells protected myocardial cells H9c2 and its protective mechanism may be achieved by promoting the autophagy of cells.
