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TMBIM6 promotes diabetic tubular epithelial cell survival and albumin endocytosis by inhibiting the endoplasmic reticulum stress sensor, IRE1α

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Abstract

Aim

Reduced albumin reabsorption in proximal tubular epithelial cells (PTECs), resulting from decreased megalin plasma membrane (PM) localization due to prolonged endoplasmic reticulum (ER) stress, potentially contributes to albuminuria in early diabetic kidney disease (DKD). To examine this possibility, we investigated the cytoprotective effect of TMBIM6 in promoting diabetic PTEC survival and albumin endocytosis by attenuating ER stress with an IRE1α inhibitor, KIRA6.

Methods and results

Renal TMBIM6 distribution and expression were determined by immunohistochemistry, western blotting, and qPCR, whereas tubular injury was evaluated in db/db mice. High-glucose (HG)-treated HK-2 cells were either treated with KIRA6 or transduced with a lentiviral vector for TMBIM6 overexpression. ER stress was measured by western blotting and ER-Tracker Red staining, whereas apoptosis was determined by performing TUNEL assays. Megalin expression was measured by immunofluorescence, and albumin endocytosis was evaluated after incubating cells with FITC-labeled albumin. Tubular injury and TMBIM6 downregulation occurred in db/db mouse renal cortical tissues. Both KIRA6 treatment and TMBIM6 overexpression inhibited ER stress by decreasing the levels of phosphorylated IRE1α, XBP1s, GRP78, and CHOP, and stabilizing ER expansion in HG-treated HK-2 cells. TUNEL assays performed with KIRA6-treated or TMBIM6-overexpressing cells showed a significant decrease in apoptosis, consistent with the significant downregulation of BAX and upregulation of BCL-2, as measured by immunoblotting. Both KIRA6 and TMBIM6 overexpression promoted megalin PM localization and restored albumin endocytosis in HG-treated HK-2 cells.

Conclusion

TMBIM6 promoted diabetic PTEC survival and albumin endocytosis by negatively regulating the IRE1α branch of ER stress.

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Data Availability

The data used to support the findings of this study are available from the corresponding author upon request.

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Acknowledgements

We extend special thanks to Dr. Wei Huili for inspiring this study and for her patience and timely help during the study.

Funding

This study was funded by National Natural Science Foundation of China (grant numbers 81774273 and 82004275).

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Authors and Affiliations

  1. Department of Nephrology and Endocrinology (A), Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China

    Huidi Xie, Yang Shi, Ying Zhou & Hongfang Liu

  2. Dongzhimen Hospital, Renal Research Institute of Beijing University of Chinese Medicine, Beijing University of Chinese Medicine, No. 5, Haiyuncang Alley, Dongcheng District, 100700, Beijing, China

    Hongfang Liu

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Xie, H., Shi, Y., Zhou, Y. et al. TMBIM6 promotes diabetic tubular epithelial cell survival and albumin endocytosis by inhibiting the endoplasmic reticulum stress sensor, IRE1α. Mol Biol Rep 49, 9181–9194 (2022). https://doi.org/10.1007/s11033-022-07744-z

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