Abstract: SA-PO305

Inhibiting Post-Translational Core Fucosylation Protects against Albumin-Induced Proximal Tubular Epithelial Cell Injury

Session Information

Category: Cell Biology

  • 204 Extracellular Matrix Biology, Fibrosis, Cell Adhesion

Author

  • Dapeng, Wang, The First Affiliated Hospital of Dalian Medical University, Dalian, China
Background

Albuminuria is an independent risk factor for renal interstitial fibrosis (RIF). Glomerular-filtered albumin may cause injury to proximal tubular epithelial cells (PTECs) in endocytic and non-endocytic pathways via megalin and TGFβRII respectively. As megalin and TGFβRII are both modified by post-translational core fucosylation which plays a critical role in RIF, we identified whether or not core fucosylation is a potential target for reducing albumin-induced injury to PTECs.

Methods

We constructed a human PTEC-derived cell line of HK-2 cells and a bovine serum albumin (BSA) injury model in vitro. RNAi was used to inhibit expression of megalin, TGFβRII and FUT8. Western blotting, immunostaining, enzyme-linked immunosorbent assay, lectin blotting and fluorescence-activated cell sorting were used to determine BSA-induced endocytic and non-endocytic damages in HK-2 cells. FUT8 is a core fucosylation-related gene, the expression of FUT8 was significantly increased after incubation with BSA in HK-2 cells.

Results

FUT8siRNA significantly reduced core fucosyaltion of megalin and TGFβRII. Meanwhile, it could also inhibit activation of the TGFβ/TGFβRII/Smad2/3 signaling pathway. Furthermore, FUT8siRNA could reduce monocyte chemotactic protein-1; reactive oxygen species and apoptosis; and significantly decrease fibronectin and collagen I levels in BSA-overloaded HK-2 cells. Notably, inhibiting core fucosylation is more effective than either inhibiting megalin or inhibiting TGFβRII in the prevention of albumin-induced injury to PTECs.

Conclusion

Inhibition of core fucosylation could effectively alleviate albumin-induced endocytic and non-endocytic injury to PTECs, our study provides a potential therapeutic target in albuminuria-induced injury.

Funding

  • Government Support - Non-U.S.