抑制核心岩藻糖基化修饰减轻嘌呤霉素氨基核苷致小鼠足细胞损伤

Objective:To investigate the effects of α1,6-fucosyltransferase 8(FUT8) on murine podocyte injury caused by puromycin aminonucleoside(PAN). Methods: A conditionally immortalized mouse podocyte clonal cell line (mouse podocyte clone 5, MPC5) was cultured in medium containing mouse γ-interferon at 33 C in vitro,FUT8siRNA suppress the expression of FUT8 in podocytes. the expression of FUT8mRNA was detected by RT-PCR, the expression of core fucose and podocyte marker-Nephrin was detected by immunofluorescent analysis, the expression of FUT8 and Nephrin was detected by western-blotting, and the apoptosis was examined by flow cytometry. Results:1. It was confirmed by immunofluorescence that core fucose existed in MPC5. 2. immunofluorescent analysis showed that the expression of core fucose was significantly increased after PAN incubation, while Nephrin was decreased; western- blotting showed that the expression of FUT8 was significantly increased, while Nephrin was decreased after PAN incubation; the apoptosis of cells was increased after PAN incubation. Preincubation with FUT8siRNA could inhibit cells detachment from the bottom of the culture dish;the expression of core fucose and FUT8 was decreased, while Nephrin was increased; the apoptosis of cells was decreased.Conclusion: 1. The core fucosylation involved in the apoptosis and the change of structural protein in podocyte injury caused by puromycin aminonucleoside.2. Utilizing RNAi technology to silence the expression of FUT8 gene , blocking of the core fucosylation, led to inhibition of podocyte injury induced by puromycin aminonucleoside.