Exploring RNA interference as a therapeutic strategy for renal disease
Exploring RNA interference as a therapeutic strategy for renal disease
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DOI：
10.1038/sj.gt.3302480
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发表时间：
2005-06-01
期刊：
影响因子：
5.1
通讯作者：
Imai, E
中科院分区：
文献类型：
--
作者：
Takabatake, Y;Isaka, Y;Imai, E
短的合成干扰RNA双链体（siRNA）可以选择性地抑制哺乳动物体细胞中的基因表达，而没有双链RNA（dsRNA）的非选择性毒性作用。然而，siRNA转移的选择性体内递送尚未在肾脏中报道。在这里，我们研究了通过肾动脉注射合成的siRNA，然后电穿孔是否可以有效和治疗性地沉默肾小球中的特定基因。我们研究了siRNA在大鼠培养的系膜细胞（MCs）中的作用，结果表明siRNA序列特异性抑制转基因表达的作用比反义寡核苷酸（ASODN）强1000倍以上。将靶向荧光素酶的siRNA转染到大鼠肾脏中显著抑制共转染的荧光素酶表达载体在体内的表达。在转基因“绿色”大鼠中递送靶向增强型绿色荧光蛋白（EGFP）的siRNA降低了内源性EGFP表达，主要是在肾小球MC中。针对TGF-β 1的RNAi可显著抑制TGF-β 1 mRNA和蛋白的表达，从而改善肾小球肾炎基质扩张的进程。此外，基于载体的RNAi在体外和体内也抑制TGF-β 1的表达。结论：siRNA介导的TGF-β 1沉默可能对纤维化疾病的预防和治疗具有重要的临床价值。
The short synthetic interfering RNA duplexes (siRNAs) can selectively suppress gene expression in somatic mammalian cells without nonselective toxic effects of double-stranded RNA ( dsRNA). However, a selective in vivo delivery of siRNA transfer has not been reported in kidney. Here, we investigated whether injection of synthetic siRNAs via renal artery followed by electroporation could be effective and therapeutic in silencing specific gene in glomerulus. We investigated the effect of siRNA in rat cultured mesangial cells (MCs) and showed that siRNA sequence-specific suppression of transgene expression was over a 1000-fold more potent than that by antisense oligodeoxynucleotide (ASODN). Transfection of siRNA targeting luciferase into rat kidneys significantly inhibited expression of a cotransfected luciferase expression vector in vivo. The delivery of siRNA targeting enhanced green fluorescent protein (EGFP) in the transgenic 'green' rat reduced endogenous EGFP expression, mainly in glomerular MCs. Furthermore, RNAi targeting against TGF-β 1 significantly suppressed TGF-β 1 mRNA and protein expression, thereby ameliorated the progression of matrix expansion in experimental glomerulonephritis. In addition, vector-based RNAi also inhibited TGF-β 1 expression in vitro and in vivo. In conclusion, siRNA-directed TGF-β 1 silencing may be of therapeutic value in the prevention and treatment of fibrotic diseases.