Lentivirus-mediated LOX-1-RNA interference attenuates oxidative stress-induced apoptosis in myocardial cells.
- Author:
Bin LIU
1
;
Jiacheng HUANG
;
Yingchun ZHOU
;
Xuegang SUN
;
Wenbi ZENG
;
Xing LI
;
Shuwen ZANG
;
Wenwen HAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis; physiology; Cells, Cultured; Genetic Vectors; genetics; Hydrogen Peroxide; Lentivirus; genetics; Myocytes, Cardiac; cytology; Oxidative Stress; RNA Interference; RNA, Small Interfering; genetics; Rats; Scavenger Receptors, Class E; genetics
- From: Journal of Southern Medical University 2012;32(2):165-168
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a lectin-like oxidized low-density lipoprotein receptor (LOX-1) RNA interference (RNAi) lentivirus and explore the role of LOX-1 in H(2)O(2)-induced apoptosis of rat myocardial cells.
METHODSLOX-1 shRNA sequence was synthesized and cloned into pLentiLox3.7 (pLL3.7) lentiviral vector to construct the lentiviral vector pLL3.7-LOX1. The lentiviral vectors (pLL3.7 and pLL3.7-LOX1) and the packaging vectors dR8.9 and VSVG were co-transfected into 293FT cells for packaging the lentivirus. H9C2 myocardial cells were infected by the lentiviruses to observe the inhibitory rate of LOX-1 on H(2)O(2)-induced apoptosis of H9C2 cells by RT-PCR, CCK-8, and Hochest33258 staining.
RESULTSDouble restriction enzyme digestion and DNA sequencing confirmed correct insertion of the sequence. Suppression of LOX-1 by the lentivirus attenuated H(2)O(2)-induced cell viability reduction and apoptosis in the myocardial cells (P<0.01).
CONCLUSIONLOX-1 activation may play an important role in H(2)O(2)-induced apoptosis of rat myocardial cells.
