Randall's plaque (RP) serves as a nidus on which idiopathic calcium oxalate stones form. Renal interstitial mineralization may be the cause underlying RP, and recent studies demonstrated the similarities between the interstitial mineralization and ectopic calcification. The present study aimed to investigate whether human renal interstitial fibroblasts (hRIFs) could form calcification under osteogenic conditions, and whether long non-coding RNA H19 participated in regulating osteogenic differentiation of hRIFs through Wnt-β-catenin pathway. HRIFs were isolated and induced for osteogenic differentiation under osteogenic conditions. Runx2, OCN, alkaline phosphatase (ALP) activity, and the mineralized nodule formation were used to assess the osteogenic phenotype. Molecule expressions were determined by qRT-PCR, immunofluorescence staining, and western blot. The mineralized nodules were assessed by Alizarin red staining. Compared to the normal renal papillary tissue, Runx2, OCN, and H19 were significantly upregulated in RP. After hRIFs were induced with osteogenic medium, osteogenic markers (Runx2, OCN and ALP), β-catenin and H19 were significantly upregulated, and the mineralized nodules are formed. Additionally, overexpression of H19 promoted the osteogenic phenotype of hRIFs and increased the expression of β-catenin, whereas knock-down of H19 or XAV939 (inhibitor of Wnt-β-catenin signaling pathway) significantly repressed the osteogenic phenotype of hRIFs and decreased the β-catenin. Moreover, XAV939 was shown to abolish the osteogenic differentiation of hRIFs promoted by H19. The study demonstrated that ectopic calcification partly participated in the formation of RP, and H19 promoted osteogenic differentiation of hRIFs by activating Wnt-β-catenin pathway, which shed new light on the molecular mechanism of the RP formation.

中文翻译：

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长的非编码RNA H19通过Wnt-β-catenin途径促进肾间质成纤维细胞的成骨分化。

兰德尔斑块（RP）可作为病原体，在其上形成特发性草酸钙结石。肾间质矿化可能是RP的根本原因，最近的研究表明间质矿化和异位钙化之间存在相似性。本研究旨在探讨在成骨条件下人肾间质成纤维细胞（hRIFs）是否能形成钙化，以及长的非编码RNA H19是否通过Wnt-β-catenin途径参与调节hRIFs的成骨分化。分离HRIF并在成骨条件下诱导成骨分化。使用Runx2，OCN，碱性磷酸酶（ALP）活性和矿化的结节形成来评估成骨表型。分子表达通过qRT-PCR，免疫荧光染色，和蛋白质印迹。通过茜素红染色评估矿化的结节。与正常肾乳头组织相比，Runx2，OCN和H19在RP中显着上调。用成骨培养基诱导hRIFs后，成骨标记物（Runx2，OCN和ALP），β-catenin和H19被显着上调，并形成矿化的结节。此外，H19的过表达促进了hRIF的成骨表型并增加了β-catenin的表达，而敲低H19或XAV939（Wnt-β-catenin信号通路的抑制剂）则显着抑制了hRIF的成骨表型并降低了β -catenin。此外，XAV939被证明可以消除H19促进的hRIF的成骨分化。研究表明，异位钙化部分参与了RP的形成，