1.Lentivirus-mediated LOX-1-RNA interference attenuates oxidative stress-induced apoptosis in myocardial cells.
Bin LIU ; Jiacheng HUANG ; Yingchun ZHOU ; Xuegang SUN ; Wenbi ZENG ; Xing LI ; Shuwen ZANG ; Wenwen HAO
Journal of Southern Medical University 2012;32(2):165-168
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.
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