Department of Urology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China (mainland).
Department of Urology, The First People's Hospital of Taicang, Taicang, Jiangsu, China (mainland).
Ann Transplant. 2021 Mar 19;26:e930066. doi: 10.12659/AOT.930066.
BACKGROUND Klotho deficiency has been implicated in various kidney diseases and has been associated with renal fibrosis. However, the role of Klotho in renal allograft fibrosis still remains undetermined. MATERIAL AND METHODS A 24-week-old rat renal transplant model with chronic allograft dysfunction (CAD) was carried out by orthotopic kidney transplantation using F344 donor rats to Lewis recipient rats. Successful establishment of the model was verified by HE and Masson staining and renal allograft function assessment. HK-2 cells were cultured and treated with TGF-ß1 and/or siRNA-Klotho at various time points. Total proteins and RNA were extracted from the cultured cells and kidney tissues. Western blot assay and quantitative RT-PCR were used to analyze the expression of Klotho, fibronectin, and ß-catenin pathways. RESULTS We successfully established and identified a 24-week-old rat renal transplant model with CAD. Loss of Klotho was identified to be associated with epithelial-to-mesenchymal transition (EMT), renal allograft fibrosis, and CAD. In HK-2 cells, a significant decrease of Klotho protein was observed in the renal fibrosis induced by TGF-ß1 in a time-dependent manner. Moreover, intervention of siRNA-Klotho remarkably promoted the progression of renal fibrosis and activation of the Wnt/ß-catenin signaling pathway. CONCLUSIONS Our results show that Klotho has a significant protective role against EMT, renal allograft fibrosis, and CAD following kidney transplantation, which is mediated by inhibition of the Wnt/ß-catenin signaling pathway.
Klotho 缺乏与各种肾脏疾病有关，并与肾纤维化有关。然而，Klotho 在肾移植纤维化中的作用仍未确定。
通过使用 F344 供体大鼠对 Lewis 受体大鼠进行原位肾脏移植，建立了 24 周龄大鼠慢性移植物功能障碍（CAD）肾移植模型。通过 HE 和 Masson 染色以及肾移植功能评估验证模型的成功建立。HK-2 细胞在不同时间点用 TGF-β1 和/或 siRNA-Klotho 处理，并从培养的细胞和肾脏组织中提取总蛋白和 RNA。Western blot 检测和定量 RT-PCR 用于分析 Klotho、纤连蛋白和 β-连环蛋白通路的表达。
我们成功建立并鉴定了 24 周龄大鼠 CAD 肾移植模型。发现 Klotho 的丢失与上皮间质转化（EMT）、肾移植纤维化和 CAD 有关。在 HK-2 细胞中，TGF-β1 在时间依赖性方式诱导的肾纤维化中观察到 Klotho 蛋白的显著减少。此外，siRNA-Klotho 的干预显著促进了肾纤维化的进展和 Wnt/β-连环蛋白信号通路的激活。
我们的结果表明，Klotho 在肾移植后对 EMT、肾移植纤维化和 CAD 具有显著的保护作用，这是通过抑制 Wnt/β-连环蛋白信号通路介导的。
