Hwang Meeyul, Kim Hye-Jin, Noh Hey-Jeong, Chang Young-Chae, Chae Young-Mi, Kim Kyung-Hyun, Jeon Jae-Pil, Lee Tae-Sung, Oh Hoon-Kyu, Lee Yun-Sik, Park Kwan-Kyu
Department of Pathology, Catholic University of Daegu School of Medicine, Daegu, Republic of Korea.
Exp Mol Pathol. 2006 Aug;81(1):48-54. doi: 10.1016/j.yexmp.2005.11.005. Epub 2006 Jan 27.
TGF-beta1 has been known as an important factor in tubulointerstitial fibrosis which is a common process in most progressive renal diseases. We hypothesized that the interstitial fibrosis could be prevented by abolishing TGF-beta1 function in unilateral ureteral obstruction (UUO)-induced renal fibrosis. shRNA vectors were generated to suppress TGF-beta1 expression at a high glucose concentration which allowed the maximal induction of TGF-beta1 in primary rat mesangial cells. An shRNA vector, designated shTB1d, significantly suppressed TGF-beta1 in both transcriptional and translational levels in vitro cultured cells and in vivo fibrosis-induced mouse kidney, accompanied by the suppression of target genes (e.g., type I collagen and PAI-1) of TGF-beta1. Furthermore, the shTB1d suppressed the expression of TGF-beta1 and type I collagen in tubulointerstitial cells until day 7 after UUO-induced fibrosis, but none- or vector-treated mice maintained their expression, suggesting that the TGF-beta1 shRNA delays the process of renal fibrosis in UUO mouse model. This work would provide a valuable tool to prevent tubulointerstitial fibrosis using RNA interference strategy.
转化生长因子β1（TGF-β1）是肾小管间质纤维化的一个重要因素，而肾小管间质纤维化是大多数进行性肾脏疾病中的常见过程。我们推测，通过消除单侧输尿管梗阻（UUO）诱导的肾纤维化中TGF-β1的功能，可以预防间质纤维化。构建了短发夹RNA（shRNA）载体，以在高葡萄糖浓度下抑制TGF-β1的表达，高葡萄糖浓度可使原代大鼠系膜细胞中TGF-β1的诱导达到最大程度。一种名为shTB1d的shRNA载体在体外培养细胞和体内纤维化诱导的小鼠肾脏中，在转录和翻译水平上均显著抑制了TGF-β1，并伴随着TGF-β1靶基因（如I型胶原和纤溶酶原激活物抑制剂-1）的抑制。此外，在UUO诱导纤维化后直到第7天，shTB1d抑制了肾小管间质细胞中TGF-β1和I型胶原的表达，但未处理或载体处理的小鼠维持了它们的表达，这表明TGF-β1 shRNA延缓了UUO小鼠模型中肾纤维化的进程。这项工作将为使用RNA干扰策略预防肾小管间质纤维化提供一个有价值的工具。
