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Mutation of Beclin1 acetylation site at K414 alleviates high glucose-induced podocyte impairment in the early stage of diabetic nephropathy by inhibiting hyperactivated autophagy

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Abstract

Background

Our group recently reported that a mutation of the novel Beclin1 K414R acetylation site impacts the stability of Beclin1 protein, which decreases autophagy in adipocytes and further impedes adipocyte differentiation and lipolysis. This study was to explore whether Beclin1 acetylation plays a role in the early renal injury induced by high glucose and to further investigate the K414R mutation site in podocytes.

Methods

Male Sprague–Dawley rats were randomized to con (control) and diabetic nephropathy (DN) groups. The DN group was induced by a single 55 mg/kg intraperitoneal injection of streptozotocin and fed a high-fat and high-sugar diet (the con group received an equal volume of the vehicle and fed a plain diet), after 3 days of induction, blood glucose levels were measured to confirm the onset of diabetes. Then, at weeks 0 and 4, the biochemical index was assayed and renal cortex tissues were harvested. MPC5 podocytes were cultured in vitro. Beclin1 (K414R)-pLVX-ZsGreen1-N1(wild-type or mutant) lentiviral plasmids were transfected into podocytes. Western blot or immunoprecipitation was used to test proteins or the acetylation levels respectively, and immunohistochemistry was used to analyze morphological changes of podocytes. Immunofluorescence was used to detect the aggregation of LC3 puncta.

Results

The acetylation level of Beclin1 was upregulated with podocyte injury exacerbated in high glucose at 24 h and that a mutation at K414R could inhibit hyperactivated autophagy, which ameliorated podocyte impairment.

Conclusion

These findings suggest that the acetylation site at K414 is a critical molecule and drug target and that further research into this area is warranted.

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Data availability

Data are available from the corresponding author on reasonable request.

Material availability

Data are available from the corresponding author on reasonable request.

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Acknowledgements

This study was supported by grants from the National Natural Science Fund Youth Project of China (No. 81600684 and No. 81603476), as well as the Youth Fund Project of the Shangdong Natural Science Foundation (ZR2016HQ16). We thank Professor Ying Yang from the Shanghai Key Laboratory of Diabetes for scientific advice.

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Authors and Affiliations

  1. Department of Endocrinology and Metabolism, First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, 215006, Jiangsu, China

    Jun Xu & Bimin Shi

  2. Department of Endocrinology and Metabolism, The Affiliated Hospital of Qingdao University, No. 1677 Wutaishan Road, Qingdao, 266000, Shandong, China

    Yujie Deng, Ping Wang, Qing Yu & Chengqian Li

  3. Department of Geriatrics, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, No. 600, Yishan Road, Shanghai, 200233, China

    Yingying Ke & Yunxia Zhu

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  1. Jun Xu
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Contributions

All authors took responsibility for all aspects of the reliability and freedom from bias of the data presented and their discussion. BS designed the study. JX, YD, and PW analyzed the results, and YK, YZ and QY wrote and edited the manuscript. BS and CL reviewed and edited the manuscript. JX, YD and WP reviewed and revised the manuscript.

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Correspondence to Bimin Shi.

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The authors declare that they have no competing interests.

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The experimental protocol was approved by the Ethics Committee for Animal Experimentation of The Affiliated Hospital of Qingdao University.

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Xu, J., Deng, Y., Ke, Y. et al. Mutation of Beclin1 acetylation site at K414 alleviates high glucose-induced podocyte impairment in the early stage of diabetic nephropathy by inhibiting hyperactivated autophagy. Mol Biol Rep 49, 3919–3926 (2022). https://doi.org/10.1007/s11033-022-07242-2

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