RNA干扰血管紧张素原基因表达对前脂肪细胞3T3-L1分化的影响

doi:10.13481/j.1671-587x.20150619

Abstract

Abstract:

Objective To construct the angiotensinogen (AGT)-targeting RNA interfering plasmids,and to explore the effect of AGT gene on the differentiation of preadipocytes 3T3-L1 in the mice. Methods The small hairpin RNA (AGT-shRNA) interference plasmids,with a scrambled Non-shRNA as control,were constructed on the basis of the activity of U6 promoter-driven expression vector psiRNA-U6.1/Neo and transfected into the mouse preadipocytes 3T3-L1.Two cell lines were generated from stable transfections.RT-PCR and SDS-PAGE were performed to monitor the mRNA and protein expressions of AGT gene, respectively.The effect of adipose AGT on the differentiation of 3T3-L1 was investigated through microscopic observation and profiling of adipocyte differentiation markers. Results Both AGT-shRNA and Non-shRNA interference vectors were successfully constructed; the expression of AGT mRNA was inhibited(P<0.05)and the expression of AGT protein was 43.86% of control.Both adipocyte differentiation markers,intracytoplasmic lipid levels and glycerol-3-phophate dehydrogenase (GPDH) activities were increased during the differentiation of preadipocytes 3T3-L1,but the data were lower than those in control group(P<0.05). Conclusion AGT gene silencing can significantly inhibit the differentiation of preadipocytes 3T3-L1,which demonstrates that there is important relationship between adipocyte metabolism and renin-angiotensin system (RAS) in adipose tissue.

Key words: angiotensinogen, RNA interference, 3T3-L1, differentiation

CLC Number:

HUANG Zhili, ZHANG Lijun, WU Lihua, WANG Yan, YANG Rude. Influence of angiotensinogen gene silencing on differentiation of preadipocytes 3T3-L1[J].Journal of Jilin University Medicine Edition, 2015, 41(06): 1195-1200.

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Influence of angiotensinogen gene silencing on differentiation of preadipocytes 3T3-L1

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