Kawase Kengo, Hamamoto Shuzo, Unno Rei, Taguchi Kazumi, Okada Atsushi, Yasui Takahiro
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences, 1, Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, Japan.
Urolithiasis. 2024 Dec 26;53(1):16. doi: 10.1007/s00240-024-01677-x.
The early stages of kidney crystal formation involve inflammation and hypoxia-induced cell injury; however, the role of the hypoxic response in kidney crystal formation remains unclear. This study investigated the effects of a prolyl hydroxylase domain inhibitor (roxadustat) on renal calcium oxalate (CaOx) crystal formation through in vitro and in vivo approaches. In the in vitro experiment, murine renal tubular cells (RTCs) were exposed to varying roxadustat concentrations and CaOx crystals. CaOx monohydrate (COM) crystal adhesion was evaluated using fluorescent labels, whereas western blotting was used to examine protein expression. Quantitative real-time polymerase chain reaction was used to analyze gene expression changes. Macrophage responses were investigated by co-culturing them with RTCs treated with COM. In the in vivo experiment, C57BL/6J mice were injected with roxadustat or saline for 2 days, followed by glyoxylate for 6 days to induce renal crystal deposition. Biochemical measurements recorded plasma erythropoietin, urinary data, and pH levels. Roxadustat suppressed the adhesion of COM crystals to RTCs and the expression of proinflammatory genes, such as chemokine (C-C motif) ligand 2 (Ccl2) and secreted phosphoprotein 1 (Spp1). Roxadustat decreased the expression levels of Ccl2, tumor necrosis factor (Tnf), and interleukin 6 (Il6) in co-cultured macrophages. In the in vivo experiment, the amount of renal CaOx crystal deposits was significantly lower in the roxadustat-treated group than in the vehicle group. Roxadustat treatment decreased Ccl2, Tnf, and adheision G protein-coupled receptor E1 (Adgre1) expression in the kidneys. Roxadustat reduced kidney inflammation and CaOx crystal deposition, suggesting its potential as a therapeutic option for kidney stone prevention.
肾脏晶体形成的早期阶段涉及炎症和缺氧诱导的细胞损伤;然而,缺氧反应在肾脏晶体形成中的作用仍不清楚。本研究通过体外和体内方法研究了脯氨酰羟化酶结构域抑制剂(罗沙司他)对肾草酸钙(CaOx)晶体形成的影响。在体外实验中,将小鼠肾小管细胞(RTCs)暴露于不同浓度的罗沙司他和CaOx晶体中。使用荧光标记评估一水合草酸钙(COM)晶体的粘附情况,而蛋白质印迹法用于检测蛋白质表达。定量实时聚合酶链反应用于分析基因表达变化。通过将巨噬细胞与经COM处理的RTCs共培养来研究巨噬细胞反应。在体内实验中,给C57BL/6J小鼠注射罗沙司他或生理盐水2天,随后注射乙醛酸6天以诱导肾脏晶体沉积。生化测量记录血浆促红细胞生成素、尿液数据和pH水平。罗沙司他抑制COM晶体与RTCs的粘附以及促炎基因的表达,如趋化因子(C-C基序)配体2(Ccl2)和分泌磷蛋白1(Spp1)。罗沙司他降低了共培养巨噬细胞中Ccl2、肿瘤坏死因子(Tnf)和白细胞介素6(Il6)的表达水平。在体内实验中,罗沙司他治疗组的肾CaOx晶体沉积物量显著低于载体组。罗沙司他治疗降低了肾脏中Ccl2、Tnf和粘附G蛋白偶联受体E1(Adgre1)的表达。罗沙司他减轻了肾脏炎症和CaOx晶体沉积,表明其作为预防肾结石治疗选择的潜力。