Effects of Inhibiting Necroptosis on H9c2 Cardiomyocytes Injury Induced by Hypoxia/Reoxygenation.
- Author:
Lihui LU
;
Mingyue ZHAO
;
Siyuan WU
;
Wenchao WU
;
Hua FU
;
Xiaojing LIU
- Publication Type:Journal Article
- MeSH:
Animals;
Apoptosis;
Cell Hypoxia;
Cell Line;
Gene Expression;
Myocytes, Cardiac;
pathology;
Protein-Serine-Threonine Kinases;
genetics;
metabolism;
RNA, Small Interfering;
Rats;
Real-Time Polymerase Chain Reaction;
Receptor-Interacting Protein Serine-Threonine Kinases;
genetics;
metabolism;
Transfection
- From:
Journal of Biomedical Engineering
2015;32(2):393-399
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study is to construct specific shRNA expressing plasmids, and to observe their effects on H9c2 cardiomyocytes injury induced by hypoxia/reoxygenation (H/R). RIPK1 and RIPK3 are the key kinases mediating the process of necroptosis. Using recombinant DNA technology, we inserted the synthetic shRNA into pSUPER vector to construct RIPK1-shRNA or RIPK3-shRNA plasmid respectively. We transfected H9c2 cardiomyocytes with the two shRNA plasmids respectively, before we treated them with H/R stimulation. Then, we measured the relevant genes and proteins by real-time PCR and Western blot. Meanwhile,we detected the markers of necroptosis and cardiomyocytes injury. The results showed that inhibition of ripk1 or ripk3 gene expression by its specific shRNA might protect the cardiomyocytes injury induced by H/R stimulation.
