Suppression of renal tubulointerstitial fibrosis by small interfering RNA targeting heat shock protein 47
Suppression of renal tubulointerstitial fibrosis by small interfering RNA targeting heat shock protein 47
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DOI：
10.1159/000108759
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发表时间：
2008-01-01
影响因子：
4.2
通讯作者：
Kohno, Shigeru
中科院分区：
文献类型：
--
作者：
Xia, Zhiyin;Abe, Katsushige;Kohno, Shigeru
背景/目的：单侧输尿管梗阻（UUO）是一种成熟的肾小管间质纤维化模型。在肾小管间质纤维化的进展过程中，在肾小管间质区域观察到胶原合成的上调和随后的胶原积累。热休克蛋白47（HSP 47）是一种胶原特异性分子伴侣，在调节胶原合成中起重要作用。我们设计了针对HSP 47 mRNA的小干扰RNA（siRNA）序列，以检测HSP 47是否参与小鼠UUO模型中肾小管间质纤维化的进展。方法：在制备UUO时经输尿管一次性注射HSP 47 siRNA。我们还应用了一种新的基因传递系统的siRNA使用阳离子化明胶微球。分别于术后7天和14天取肾。HSP 47和I型，III型和IV型胶原蛋白的表达水平进行了分析，通过免疫组化和蛋白质印迹。结果如下：UUO后7天，HSP 47和I、III、IV型胶原的表达水平在梗阻肾或绿色荧光蛋白siRNA处理的梗阻肾中显著上调。注射HSP 47 siRNA显著降低了蛋白表达水平，并显著减轻了伴随的间质纤维化。此外，阳离子化明胶微球作为一个传递系统增强和延长HSP 47 siRNA的抗纤维化作用。结论：我们的研究结果表明，热休克蛋白47是一个候选的目标，用于预防肾小管间质纤维化和选择性阻断热休克蛋白47的表达，通过使用siRNA可能是一个潜在的有用的治疗方法，肾脏疾病患者。版权所有（c）2007 S. Karger AG，巴塞尔。
Background/Aim: Unilateral ureteral obstruction (UUO) is a well-established model for tubulointerstitial fibrosis. During the progression of tubulointerstitial fibrosis, upregulation of collagen synthesis and subsequent accumulation of collagen were observed in the tubulointerstitial area. Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone and plays an essential role in regulating collagen synthesis. We designed small interfering RNA ( siRNA) sequences for HSP47 mRNA to examine whether HSP47 is involved in the progression of renal tubulointerstitial fibrosis in a mouse UUO model. Methods: The HSP47 siRNA was injected once via the ureter at the time of UUO preparation. We also applied a new gene delivery system for siRNA using cationized gelatin microspheres. The kidneys were harvested 7 and 14 days after UUO. The HSP47 and type I, III, and IV collagen expression levels were analyzed by immunohistochemistry and Western blotting. Results: Seven days after UUO, the expression levels of HSP47 and type I, III, and IV collagens were markedly upregulated in obstructed kidneys or green fluorescent protein siRNA treated obstructed kidneys. HSP47 siRNA injection significantly reduced the protein expression levels and significantly diminished the accompanying interstitial fibrosis. Moreover, cationized gelatin microspheres as a delivery system enhanced and lengthened the antifibrotic effect of HSP47 siRNA. Conclusions: Our results indicate that HSP47 is a candidate target for the prevention of tubuloin terstitial fibrosis and that selective blockade of the HSP47 expression by using siRNA could be a potentially useful therapeutic approach for patients with renal disease. Copyright (c) 2007 S. Karger AG, Basel.