Department of Nephrology, University of Chinese Academy of Sciences Shenzhen Hospital, Shenzhen, China.
Department of Nephrology, Wuxi No. 2 People's Hospital, Nanjing Medical University, Wuxi, China.
J Cell Physiol. 2019 Jun;234(6):9673-9686. doi: 10.1002/jcp.27654. Epub 2018 Dec 17.
Uremia can affect hepatic metabolism of drugs by regulating the clearance of drugs, but it has not been clarified whether gene silencing could modulate the epithelial-mesenchymal transition (EMT) process in uremia. Hence, we investigated the effect of WISP1 gene silencing on the renal tubular EMT in uremia through the wnt/β-catenin signaling pathway. Initially, microarray-based gene expression profiling of uremia was used to identify differentially expressed genes. Following the establishment of uremia rat model, serum creatinine, and urea nitrogen of rats were detected. Renal tubular epithelial cells (TECs) were transfected with shRNA-WISP1 lentivirus interference vectors and LiCI (the wnt/β-catenin signaling pathway activator) to explore the regulatory mechanism of WISP1 in uremia in relation to the wnt/β-catenin signaling pathway. Then, expression of WISP1, wnt2b, E-cadherin, α-SMA, c-myc, Cyclin D1, MMP-2, and MMP-9 was determined. Furthermore, TEC migration and invasion were evaluated. Results suggested that WISP1 and the wnt/β-catenin signaling pathway were associated with uremia. Uremic rats exhibited increased serum creatinine and urea nitrogen levels, upregulated WISPl, and activated wnt/β-catenin signaling pathway. Subsequently, WISP1 silencing decreased wnt2b, c-myc, Cyclin D1, α-SMA, MMP-2, and MMP-9 expression but increased E-cadherin expression, whereas LiCI treatment exhibited the opposite trends. In addition, WISP1 silencing suppressed TEC migration and invasion, whereas LiCI treatment promoted TEC migration and invasion. The findings indicate that WISP1 gene silencing suppresses the activation of the wnt/β-catenin signaling pathway, thus reducing EMT of renal TECs in uremic rats.
尿毒症可通过调节药物清除率影响药物的肝脏代谢，但尚未阐明基因沉默是否能调节尿毒症中的上皮-间充质转化（EMT）过程。因此，我们通过 wnt/β-catenin 信号通路研究了 WISP1 基因沉默对尿毒症肾小管 EMT 的影响。首先，我们使用基于微阵列的基因表达谱分析鉴定了尿毒症的差异表达基因。在建立尿毒症大鼠模型后，检测了大鼠的血清肌酐和尿素氮。用 shRNA-WISP1 慢病毒干扰载体和 LiCI（wnt/β-catenin 信号通路激活剂）转染肾小管上皮细胞（TEC），以探讨 WISP1 在尿毒症中与 wnt/β-catenin 信号通路的关系的调节机制。然后，测定 WISP1、wnt2b、E-钙黏蛋白、α-SMA、c-myc、Cyclin D1、MMP-2 和 MMP-9 的表达。此外，评估了 TEC 的迁移和侵袭。结果表明，WISP1 和 wnt/β-catenin 信号通路与尿毒症有关。尿毒症大鼠表现出血清肌酐和尿素氮水平升高，WISPl 上调，wnt/β-catenin 信号通路激活。随后，WISP1 沉默降低了 wnt2b、c-myc、Cyclin D1、α-SMA、MMP-2 和 MMP-9 的表达，但增加了 E-钙黏蛋白的表达，而 LiCI 处理则呈现相反的趋势。此外，WISP1 沉默抑制了 TEC 的迁移和侵袭，而 LiCI 处理促进了 TEC 的迁移和侵袭。这些发现表明，WISP1 基因沉默抑制了 wnt/β-catenin 信号通路的激活，从而减少了尿毒症大鼠肾 TEC 中的 EMT。
