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Inhibition of RhoA expression reduces the secretion of extracellular matrix components in hepatic stellate cells
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 19
- DOI:
- 10.3969/j.issn.1009-3079.2010.19.017
- Key Word:
- RhoA;反义寡核苷酸;小干扰RNA;肝纤维化;基因治疗
Abstract： 目的:观察并比较两种方法抑制RhoA的表达对细胞外基质成分,如Ⅰ型胶原、透明质酸以及层粘连蛋白合成的影响,为肝纤维化的基因治疗寻求新的途径.方法:设计并合成针对RhoA相同靶点的反义寡核苷酸和小干扰RNA.分别转染肝星状细胞株HSC-T6,逆转录PCR技术检测细胞中RhoA和Ⅰ型胶原mRNA的表达:Western blot检测细胞中RhoA蛋白质的表达;ELISA检测培养上清中透明质酸(HA)、层黏蛋白(LN)及PIIIP的含量.结果:转染RhoA反义寡核苷酸和小干扰RNA Ratl质粒后.HSC-T6中RhoA mRNA的表达分别由0.892±0.051、0.937±0.044降为0.113±0.024、0.212±0.042;蛋白质表达水平均明显下调;细胞内Ⅰ型胶原mRNA的表达分别由0.709±0.067、0.695±0.087降为0.436±0.037、0.201±0.044;细胞外基质成分如HA、LN及PIIIP的表达水平明显下降.两者相比,小干扰RNA具有更强的生物学效应.结论:靶向抑制细胞因子RhoA的表达可明显减少肝星状细胞株HSC-T6细胞外基质的合成,且小干扰RNA的效果明显优于针对同一靶点的反义寡核苷酸,显示RNAi技术应用于肝纤维化基因治疗的良好前景.
- [1]王波,王天才,梁扩寰.细胞因子RhoA在实验性大鼠肝纤维化形成过程中的作用机制[J].2004,(7).
- [2]王艳芬,田德安,王波,等.RhoA特异性RNA干扰对肝星状细胞T6细胞外基质表达的影响[J].2007,(7).
- [3]Jin,Y.,Liu,S.,Yu,B.,etc.Targeted delivery of antisense oligodeoxynucleotide by transferrin conjugated pH-sensitive lipopolyplex nanoparticles: A novel oligonucleotide- based therapeutic strategy in acute myeloid leukemia[J].2010,7(1).
- [4]Rayburn ER,Zhang R.Antisense, RNAi, and gene silencing strategies for therapy: Mission possible or impossible?[J].2008,13(11).
- [5]Valdes JJ,Bentley WE,Hebert CG.Beyond silencing--engineering applications of RNA interference and antisense technology for altering cellular phenotype[J].2008,19(5).
- [6]Rockey DC,Piantedosi R,Vogel S,etc.An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro.[J].2000,41(6).
- [7]Friedman SL.Mechanisms of hepatic fibrogenesis.[J].2008,134(6).
- [8]Nowotny, Marcin,Yang, Wei.Structural and functional modules in RNA interference[J].2009,19(3).
- [9]Du,L.,Gatti,R.A..Progress toward therapy with antisense-mediated splicing modulation[J].2009,11(2).