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Puerarin prevents calcium oxalate crystal-induced renal epithelial cell autophagy by activating the SIRT1-mediated signaling pathway

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Abstract

Calcium oxalate (CaOx) crystals can activate autophagy, causing damage to renal tubular epithelial cells (TECs). Puerarin has been shown to have protective and therapeutic effects against a variety of diseases by inhibiting autophagy activation. However, the protective effect of puerarin against CaOx crystals and the underlying molecular mechanisms are unclear. Cell Counting Kit-8 (CCK-8) assays were used to evaluate the effects of puerarin on cell viability. Intracellular reactive oxygen species (ROS) levels were measured by the cell-permeable fluorogenic probe 2,7-dichlorofluorescein diacetate (DCFH-DA). Immunofluorescence, immunohistochemistry, and western blotting were used to examine the expression of SIRT1, Beclin1, p62, and LC3, and explore the underlying molecular mechanisms in vivo and in vitro. Puerarin treatment significantly attenuated CaOx crystal-induced autophagy of TECs and CaOx cytotoxicity to TECs by altering SIRT1 expression in vitro and in vivo, whereas the SIRT1-specific inhibitor EX527 exerted contrasting effects. In addition, we found that the protective effect of puerarin was related to the SIRT1/AKT/p38 signaling pathway. The findings suggest that puerarin regulates CaOx crystal-induced autophagy by activating the SIRT1-mediated signaling pathway, and they suggest a series of potential therapeutic targets and strategies for treating nephrolithiasis.

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Funding

This work was supported by the National Natural Science Foundation of China (NFSC, No. 81370803, 81600543), the Natural Science Foundation of Chongqing (Postdoctoral Fund): cstc2021jcyj-bsh0024, the China Postdoctoral Science Foundation (2018M631049), the Heilongjiang Postdoctoral Fund (LBH-Z18122).

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  1. Guan-Hua Jing and Ya-Dong Liu have contributed equally to this work and should be considered co-first authors.

Authors and Affiliations

  1. Department of Urology, The First Affiliated Hospital of Harbin Medical University, Harbin, China

    Guan-Hua Jing, Yin-Shan Jin, Shi-Liang Yu & Rui-Hua An

  2. Institute of Ultrasound Imaging, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Ya-Dong Liu

  3. Department of Urology, Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, Chengdu, China

    Jian-Nan Liu

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G-HJ and Y-DL contributed equally to this work and should be considered co-first authors. All authors have read and approved the final manuscript.

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Jing, GH., Liu, YD., Liu, JN. et al. Puerarin prevents calcium oxalate crystal-induced renal epithelial cell autophagy by activating the SIRT1-mediated signaling pathway. Urolithiasis 50, 545–556 (2022). https://doi.org/10.1007/s00240-022-01347-w

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