Department of Pathology, Third Xiangya Hospital, Central South University, Changsha 410013, PR China.
Medical Laboratory Center, Third Xiangya Hospital, Central South University, Changsha 410013, PR China.
Life Sci. 2020 Jul 1;252:117589. doi: 10.1016/j.lfs.2020.117589. Epub 2020 Mar 24.
Renal fibrosis is the characteristic of all kinds of chronic kidney diseases (CKDs). Fascin-1 plays an important role in tumor development, but the roles of fascin-1 in renal fibrosis have not been studied. Here, we explored the role of fascin-1 in renal fibrosis and the potential mechanisms.
Kidney unilateral ureteral obstruction (UUO) mouse model was used as an in vivo model, and proximal tubule epithelial cell lines treated with TGF-β1 were used as in vitro model of renal fibrosis. Cell transfection was performed to manipulate the expression of miR-200b/c, fascin-1 and CD44. Western blotting, qRT-PCR, immunohistochemistry or immunofluorescence assays were used to measure levels of miR-200b/c, fascin-1, CD44, and fibrosis and EMT-related markers. H&E and Masson stainings were used to examine the degree of injury and fibrosis in kidneys. Dual luciferase assay was used to examine the interaction between miR-200b/c family and fascin-1.
Fascin-1 and CD44 levels were both significantly up-regulated while miR-200b/c family was reduced in models of renal fibrosis. Furthermore, overexpression of miR-200b/c family and inhibition of fascin-1 or CD44 ameliorated renal fibrosis through suppressing EMT process. Mechanistically, miR-200b/c family directly and negatively regulated the expression of fascin-1. Overexpression of fascin-1 could reverse the effects of miR-200b/c family on renal fibrosis, and fascin-1 regulated renal fibrosis by activating CD44.
Our study is the first to show that fascin-1 plays a critical role in renal fibrosis. MiR-200b/c family could inhibit renal fibrosis through modulating EMT process by directly targeting fascin-1/CD44 axis.
肾纤维化是各种慢性肾脏病（CKD）的特征。Fascin-1 在肿瘤发展中起着重要作用，但 fascin-1 在肾纤维化中的作用尚未得到研究。在这里，我们探讨了 fascin-1 在肾纤维化中的作用及其潜在机制。
采用单侧输尿管梗阻（UUO）小鼠模型作为体内模型，采用 TGF-β1 处理的近端肾小管上皮细胞系作为肾纤维化的体外模型。通过细胞转染来操纵 miR-200b/c、fascin-1 和 CD44 的表达。Western blot、qRT-PCR、免疫组化或免疫荧光检测用于测量 miR-200b/c、fascin-1、CD44 以及纤维化和 EMT 相关标志物的水平。H&E 和 Masson 染色用于检测肾脏的损伤和纤维化程度。双荧光素酶报告实验用于检测 miR-200b/c 家族与 fascin-1 之间的相互作用。
在肾纤维化模型中，fascin-1 和 CD44 水平均显著上调，而 miR-200b/c 家族则下调。此外，miR-200b/c 家族的过表达以及 fascin-1 或 CD44 的抑制可通过抑制 EMT 过程改善肾纤维化。机制上，miR-200b/c 家族可直接负调控 fascin-1 的表达。过表达 fascin-1 可逆转 miR-200b/c 家族对肾纤维化的作用，而 fascin-1 通过激活 CD44 调节肾纤维化。
本研究首次表明 fascin-1 在肾纤维化中起着关键作用。miR-200b/c 家族可通过直接靶向 fascin-1/CD44 轴调节 EMT 过程来抑制肾纤维化。
