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DNA hypermethylation of sFRP5 contributes to indoxyl sulfate-induced renal fibrosis

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Abstract

Renal fibrosis is the most common outcome of chronic kidney disease (CKD), while the pathogenesis of renal fibrosis is not fully understood. In this study, we first showed that the progress of renal fibrosis was positively related to serum levels of indoxyl sulfate, a typical protein-bound toxin, and that there was a close correlation between serum indoxyl sulfate levels and β-catenin expression in the kidneys (r = 0.908, p < 0.001) of CKD patients. We then demonstrated that intraperitoneal injections of indoxyl sulfate (100 mg/kg/day) for 4 weeks in uninephrectomized mice explicitly induced renal fibrosis, which was accompanied by a significant activation of Wnt/β-catenin signaling. In vitro investigations in human renal tubular HK-2 cells revealed that indoxyl sulfate exhibited a potent ability to induce Wnt/β-catenin activation through the downregulation of sFRP5, a gene that codes for an extracellular antagonist of Wnt signaling, by increasing the DNA methylation level of its promoter CpG islands. The increased expression of DNA methyltransferases following the activation of ROS/ERK1/2 signaling was responsible for the DNA hypermethylation of sFRP5 induced by indoxyl sulfate. Conversely, treatment with 5-aza-2′-deoxycytidine, an inhibitor of DNA methyltransferases, significantly reduced indoxyl sulfate-induced sFRP5 downregulation and Wnt/β-catenin activation. In vivo, intraperitoneal injections of recombinant sFRP5 protein or 5-aza-2′-deoxycytidine substantially alleviated renal fibrosis in indoxyl sulfate-treated uninephrectomized mice. Our results suggest that indoxyl sulfate promotes renal fibrosis through the induction of DNA hypermethylation of sFRP5, and thereafter the activation of Wnt/β-catenin signaling. These findings provide new insights into the pathogenesis of renal fibrosis in CKD patients.

Key messages

  • IS induces renal fibrosis by increasing ß-catenin expression in CKD mice.

  • IS-induced Wnt signaling activation is due to sFRP5 hypermethylation in HK-2 cells.

  • ROS/ERK1/2 signaling activation is involved in IS-induced sFRP5 hypermethylation.

  • sFRP5 upregulation attenuates IS-induced renal fibrosis by inhibiting Wnt signaling.

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Acknowledgments

This study was supported by research grants from the National Natural Science Foundation of China (Nos. 81500567, 81500544, 81270290, and 81400747) and the project for Chongqing basic science and advanced technology research (cstc2015jcyjBX0028).

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  1. Yanlin Yu and Xu Guan contributed equally to this work.

Authors and Affiliations

  1. Department of Nephrology, Institute of Nephrology of Chongqing and Kidney Center of PLA, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, People’s Republic of China

    Yanlin Yu, Xu Guan, Ling Nie, Yong Liu, Ting He, Jiachuan Xiong, Xinli Xu, Yan Li, Ke Yang, Yiqin Wang, Yunjian Huang, Bing Feng, Jingbo Zhang & Jinghong Zhao

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  1. Yanlin Yu
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  2. Xu Guan
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  3. Ling Nie
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Correspondence to Jinghong Zhao.

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The study protocol was approved by the Ethics Committee of Xinqiao Hospital and carried out in accordance with the Declaration of Helsinki.

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Yu, Y., Guan, X., Nie, L. et al. DNA hypermethylation of sFRP5 contributes to indoxyl sulfate-induced renal fibrosis. J Mol Med 95, 601–613 (2017). https://doi.org/10.1007/s00109-017-1538-0

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