Effects of antiatherosclerosis in carotid artery by RNAi-mediated silencing of MCP-1 expression.
BACKGROUND
Our objective was to identify the effects of MCP-1 siRNA in vivo transfection in an atherosclerosis model on local expression of MCP-1 and pathogenesis of atherosclerosis.


METHODS
Carotid atherosclerosis was induced in 28 New Zealand white rabbits. Rabbits were divided into three groups randomly: RNAi group, model group, and blank plasmid group. siRNA-expressing vector was transfected to blood vessels by liposomes. The carotid arteries were processed for morphological evaluation. Local expression of MCP-1 was detected by immunohistochemistry, RT-PCR, and Western blot.


RESULTS
On hematoxylin and eosin-stained sections, partial endothelial cells detached while intimae were less thickened in the RNAi group compared to the model and blank plasmid groups; the I:M ratio was significantly reduced to 1.46 in the RNAi group compared to the model and blank plasmid groups (5.55 and 5.27, respectively). The results of immunohistochemistry showed that MCP-1 expression was less colorized and less positive in the RNAi group. RT-PCR and Western blot showed reduced expression in the RNAi group than in the model and blank plasmid groups. There were highly positive correlations between semiquantitative RT-PCR and the I:M ratio (r = 0.968).


CONCLUSIONS
Expression of MCP-1 was successfully inhibited by transfecting MCP-1 siRNA expression plasmid to the carotid artery, and the progression of atherosclerosis was restricted by RNAi-mediated silencing of MCP-1 expression.
Hu
Zuo-jun
ZJ
Department of Vascular Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China. zuojunhu@hotmail.com
Liao
Yi
Y
Hu
Ling-yu
LY
Wang
Jia-li
JL
Li
Jie
J
Lu
Wei-ming
WM
Wang
Shen-ming
SM
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002339:Carotid Arteries / Q000378:metabolism* / Q000473:pathology; D002340:Carotid Artery Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D018450:Disease Progression; D015536:Down-Regulation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D018932:Chemokine CCL2; D034741:RNA, Small Interfering