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Published on: July 10, 2019
Lentiviral vector PLV-PI3KCG gene transfer inhibits hypoxic cardiomyocytes apoptosis
Yan-Yan Li1, Hui Zhang2, Xin-Zheng Lu3
1Department of Geriatrics, First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China.
Abstract:
The PI3K/Akt signal pathway was suggested to be associated with apoptosis. However, it was still unclear whether activated PI3K/Akt signaling pathway could inhibit hypoxic cardiomyocytes apoptosis. In this research, the recombinant PI3KCG lentiviral vector plasmid (PLV-PI3KCG) was constructed and transfected into neonatal rat hypoxia/reoxygenation (H/R) injury cardiomyocytes models which were randomly divided into five groups as the normal control group, H/R group, HR empty plasmid group (HRE group), HR PLV-PI3KCG transfection preconditioning group (HRP group), and HR PLV-PI3KCG transfection + LY294002 group (HRPL group). Compared with the H/R, HRE and HRPL groups, the cardiomyocytes beat frequency and survival rate in the HRP group were significantly increased (P<0.05) and the released LDH were significantly decreased (P<0.05). The Bcl-2/Bax ratio was significantly lower in H/R, HRE and HRPL groups than that in HRP group (P<0.05). Activated PI3K/Akt signaling pathway could play a protection role in the cardiomyocytes H/R injury process which could be inhibited by LY294002.
Insights
Activating the PI3K/Akt pathway protects heart cells from injury. This pathway
Area of Science:
- Cardiovascular Biology
- Cellular Signaling
- Molecular Medicine
Background:
- The PI3K/Akt signaling pathway is implicated in apoptosis.
- Its role in inhibiting hypoxic cardiomyocyte apoptosis remains unclear.
Purpose of the Study:
- To investigate the protective role of activated PI3K/Akt signaling in cardiomyocytes against hypoxia/reoxygenation (H/R) injury.
- To determine if LY294002 inhibits this protective effect.
Main Methods:
- Constructed a recombinant PI3KCG lentiviral vector (PLV-PI3KCG).
- Transfected neonatal rat cardiomyocytes with PLV-PI3KCG to model H/R injury.
- Utilized five experimental groups: control, H/R, HRE (empty plasmid), HRP (PLV-PI3KCG), and HRPL (PLV-PI3KCG + LY294002).
Main Results:
- The HRP group showed significantly increased cardiomyocyte beat frequency and survival rate compared to H/R, HRE, and HRPL groups (P<0.05).
- LDH release was significantly decreased in the HRP group (P<0.05).
- The Bcl-2/Bax ratio was significantly lower in H/R, HRE, and HRPL groups than in the HRP group (P<0.05).
Conclusions:
- Activated PI3K/Akt signaling pathway plays a protective role in cardiomyocyte H/R injury.
- LY294002, an inhibitor, can block this protective effect.
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