Influence of angiotensinogen gene silencing on differentiation of preadipocytes 3T3-L1
10.13481/j.1671-587x.20150619
- VernacularTitle: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
- Author:
Zhili HUANG
;
Lijun ZHANG
;
Lihua WU
;
Yan WANG
;
Rude YANG
- Publication Type:Journal Article
- Keywords:
angiotensinogen;
RNA interference;
3T3-L1;
differentiation
- From:
Journal of Jilin University(Medicine Edition)
2015;(6):1195-1200
- CountryChina
- Language:Chinese
-
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.
