Oncology Center, Department of Urology, Renmin Hospital of Wuhan University and State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, China.
BJU Int. 2011 Mar;107(6):1002-8. doi: 10.1111/j.1464-410X.2010.09520.x. Epub 2010 Aug 25.
• To compare the expressions of common fibrosis-relevant genes in hydronephrosis-induced fibrotic renal tissues and normal human renal tissues, thereby providing insights into the cellular and molecular mechanisms of renal fibrosis resulting from hydronephrosis.
• A total of 12 extensively fibrotic renal tissue samples from patients with hydronephrosis (H-group) and six normal renal tissue samples from patients who underwent nephrectomy for renal cell carcinoma (N-group), along with their clinical data, were collected at Renmin Hospital of Wuhan University in China between October 2005 and August 2007. • These tissue samples were compared for their transforming growth factor-β (TGF-β)/bone morphogenetic protein (BMP) pathway-related gene profiles using a real-time polymerase chain reaction (PCR) microarray. • Subsequently, reverse transcriptase-PCR assays were used to validate the expression changes of left-right determination factor (LEFTY), a gene of interest, at the mRNA level. • The different expression of LEFTY at the protein level was confirmed by western blotting and immunohistochemistry assays.
• The results showed that 49 genes were differently expressed in fibrotic renal tissues relative to normal control tissues. Among these genes, 25 were up-regulated and 24 were down-regulated. • LEFTY-B, one of the most markedly altered genes, was down-regulated 13.55-fold compared with N-group tissues. • RT-PCR showed that the LEFTY-A (6.05-fold down-regulated, P < 0.001) and LEFTY-B (12.5-fold down-regulated, P < 0.001) genes, two members of the LEFTY family in human tissues, were both significantly down-regulated in H-group tissues. • Similarly, down-regulations of LEFTY-A (0.25-fold vs N-group, P < 0.001) and LEFTY-B (0.20-fold vs N-group, P < 0.001) proteins were detected by western blotting (P < 0.001). • Immunohistochemical staining showed different distributions of LEFTY in the two tissue samples, and quantitative image analyses confirmed that LEFTY protein expression was lower in H-group tissues than in N-group tissues (P < 0.001).
• The gene and protein expressions of LEFTY were found to be down-regulated in extensively fibrotic renal tissues induced by hydronephrosis. • LEFTY may represent an ideal candidate for a therapeutic target for renal fibrosis.
• 比较肾盂积水诱导的纤维化肾组织和正常人类肾组织中常见纤维化相关基因的表达，从而深入了解肾盂积水引起的肾纤维化的细胞和分子机制。
• 2005 年 10 月至 2007 年 8 月，在中国武汉大学人民医院收集了 12 例广泛纤维化肾组织样本（H 组）和 6 例因肾细胞癌行肾切除术的正常肾组织样本（N 组），并记录了患者的临床资料。• 使用实时聚合酶链反应（PCR）微阵列比较这些组织样本中转化生长因子-β（TGF-β）/骨形态发生蛋白（BMP）通路相关基因谱。• 随后，使用逆转录-PCR 检测验证左向右决定因子（LEFTY）感兴趣基因在 mRNA 水平的表达变化。• 通过 Western 印迹和免疫组织化学检测证实 LEFTY 在蛋白质水平的不同表达。
• 结果显示，纤维化肾组织中与正常对照组织相比，有 49 个基因表达不同。在这些基因中，25 个上调，24 个下调。• LEFTY-B 是变化最显著的基因之一，与 N 组组织相比下调 13.55 倍。• RT-PCR 显示 LEFTY-A（6.05 倍下调，P < 0.001）和 LEFTY-B（12.5 倍下调，P < 0.001）基因，组织中 LEFTY 家族的两个成员，在 H 组组织中均显著下调。• 同样，通过 Western 印迹（P < 0.001）检测到 LEFTY-A（与 N 组相比 0.25 倍下调，P < 0.001）和 LEFTY-B（与 N 组相比 0.20 倍下调，P < 0.001）蛋白的下调。• 免疫组织化学染色显示 LEFTY 在两种组织样本中的分布不同，定量图像分析证实 LEFTY 蛋白表达在 H 组组织中低于 N 组组织（P < 0.001）。
• 肾盂积水诱导的广泛纤维化肾组织中 LEFTY 的基因和蛋白表达下调。• LEFTY 可能是肾纤维化治疗靶点的理想候选者。
