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Autophagy inhibitors promoted aristolochic acid I induced renal tubular epithelial cell apoptosis via mitochondrial pathway but alleviated nonapoptotic cell death in mouse acute aritolochic acid nephropathy model

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Abstract

Aristolochic acid I (AAI) can induce renal tubular epithelial cells (RTECs) autophagy, which thereby extenuates apoptosis in vitro. In this study, we aimed to determine whether the in vitro data also apply to the AAI-induced pathologic condition in vivo. BALB/c mice were treated with AAI, autophagy inhibitors [3-methyladenine (3MA) or chloroquine diphosphate salt (CQ)], and AAI plus the inhibitors for consecutive 5 days, respectively. Mice were euthanized on day 3 and 5. AAI induced RTECs autophagy was confirmed by electron microscopy and western blot. The results showed induction of apoptotic RTECs and up-regulation of mitochondrial and endoplasmic reticulum stress-related proteins in AAI-treated mice at both of the two time points. There were more apoptotic RTECs in AAI + inhibitor groups, which might be due to increased mitochondrial stress-related proteins (cytochrome C and apoptotic protease activating factor 1, APAF-1). On day 5, severe tubulointerstitial injuries induced by AAI led to a significant decline in kidney function. There were numerous autolysosomes in dying RTECs of the AAI group. Autophagy inhibitors increased AAI-induced RTECs mitochondrial apoptosis by increasing mitochondrial stress-related proteins, but they partially mitigated the AAI-induced severe renal tubulointerstitial injury. These results confirmed that AAI could induce autophagy in RTECs, which prevented apoptosis via mitochondrial pathway in vivo. However, continuous stimulation with AAI induced excess autophagy, which ultimately resulted in AAI-induced cell death. It suggested that apoptosis wasn’t the main culprit in acute aristolochic acid nephropathy mice model.

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Acknowledgments

This work was supported by Chinese National Natural Science Research Grants to Prof. Xiao Yang (Grant numbers: 81073138) and postdoctoral fellow Youjia Zeng (Grant numbers: 81202671), respectively.

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Authors and Affiliations

  1. Department of Nephrology, The First Affiliated Hospital, Sun Yat-sen University, Key Laboratory of Nephrology, Ministry of Health, Guangzhou, 510080, Guangdong, China

    Youjia Zeng, Juan Wu, Wenfang Chen, Wenxing Peng, Xueqing Yu & Xiao Yang

  2. Department of Nephrology, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, 518033, Guangdong, China

    Youjia Zeng, Shunmin Li & Huili Sun

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  1. Youjia Zeng
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  2. Shunmin Li
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  3. Juan Wu
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  6. Wenxing Peng
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Correspondence to Xiao Yang.

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Zeng, Y., Li, S., Wu, J. et al. Autophagy inhibitors promoted aristolochic acid I induced renal tubular epithelial cell apoptosis via mitochondrial pathway but alleviated nonapoptotic cell death in mouse acute aritolochic acid nephropathy model. Apoptosis 19, 1215–1224 (2014). https://doi.org/10.1007/s10495-014-0996-x

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  • DOI: https://doi.org/10.1007/s10495-014-0996-x

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