Department of Urology, Second Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, China.
Department of Urology, Second Affiliated Hospital of Xinjiang Medical University, Urumqi, 830036, China.
Int Urol Nephrol. 2023 Feb;55(2):263-276. doi: 10.1007/s11255-022-03398-9. Epub 2022 Nov 7.
Oxidative damage is important in calcium oxalate (CaOx) stone development but occurs via multiple pathways. Studies have shown that klotho plays an essential role in ameliorating oxidative damage. This study aims to explore the role of klotho in CaOx stones and whether the underlying mechanism is related to the regulation of Keap1-Nrf2-ARE signaling.
The levels of GSH, SOD, CAT, MDA, and ROS were examined by ELISA. The klotho, Bcl-2, caspase-3, Keap1, Nrf2, HO-1, and NQO1 mRNA levels were measured by qRT‒PCR, and their protein levels were detected by Western blotting. Renal tissue apoptosis was examined by TUNEL staining, and crystal cell adherence and apoptosis in HKC cells were assessed based on the Ca concentrations and by flow cytometry. The renal pathological changes and the adhesion of CaOx crystals in the kidneys were examined by hematoxylin-eosin and von Kossa staining, respectively.
We constructed a CaOx kidney stone model in vitro. By regulating the klotho gene, klotho overexpression inhibited the CaOx-induced promotion of crystal cell adherence and apoptosis in HKC cells, and these effects were reversed by klotho knockdown. Moreover, our in vivo assay demonstrated that klotho overexpression alleviated glyoxylate administration-induced renal oxidative damage, renal apoptosis, and crystal deposition in the kidneys of mice, and these effects were also associated with activation of the Keap1-Nrf2-ARE pathway.
Klotho protein inhibits the oxidative stress response of HKC cells through the Keap1-Nrf2-ARE signaling pathway, reduces the apoptosis of and adhesion of CaOx crystals to HKC cells, and decreases the occurrence of CaOx kidney stones.
氧化损伤在草酸钙（CaOx）结石形成中很重要，但它通过多种途径发生。研究表明 klotho 在减轻氧化损伤方面起着至关重要的作用。本研究旨在探讨 klotho 在 CaOx 结石中的作用，以及潜在机制是否与 Keap1-Nrf2-ARE 信号通路的调节有关。
通过 ELISA 检测 GSH、SOD、CAT、MDA 和 ROS 的水平。通过 qRT-PCR 测量 klotho、Bcl-2、caspase-3、Keap1、Nrf2、HO-1 和 NQO1 的 mRNA 水平，并通过 Western blot 检测其蛋白水平。通过 TUNEL 染色检测肾组织细胞凋亡，通过流式细胞术根据 Ca 浓度评估 HKC 细胞中晶体细胞黏附和凋亡。通过苏木精-伊红和 von Kossa 染色分别检查肾组织的病理变化和肾脏中 CaOx 晶体的黏附。
我们在体外构建了 CaOx 肾结石模型。通过调节 klotho 基因，klotho 过表达抑制了 CaOx 诱导的 HKC 细胞晶体细胞黏附和凋亡的促进作用，而 klotho 敲低则逆转了这些作用。此外，我们的体内实验表明，klotho 过表达减轻了甘氨酸氧化酶给药诱导的小鼠肾氧化损伤、肾细胞凋亡和肾脏中 CaOx 晶体沉积，这些作用也与 Keap1-Nrf2-ARE 通路的激活有关。
klotho 蛋白通过 Keap1-Nrf2-ARE 信号通路抑制 HKC 细胞的氧化应激反应，减少 CaOx 晶体对 HKC 细胞的凋亡和黏附，降低 CaOx 肾结石的发生。
20220304。
