Apolipoprotein L1 (APOL1) wild type (G0) plays a role in the metabolism of sphingolipids, glycosphingolipids, sphingomyelin and ceramide, which constitute bioactive components of the lipid rafts (DRM). We asked whether APOL1 variants (APOL1-Vs) G1 and G2 carry the potential to alter the metabolism of sphingolipids in human podocytes. The sphingolipid pattern in HPs overexpressing either APOL1G0 or APOL1-Vs was analysed by using a thin mono- and bi-dimensional layer chromatography, mass-spectrometry and metabolic labelling with [1-³H]sphingosine. HP G0 and G1/G2-Vs exhibit a comparable decrease in lactosylceramide and an increase in the globotriaosylceramide content. An analysis of the main glycohydrolases activity involved in glycosphingolipid catabolism showed an overall decrease in the activeness of the tested enzymes, irrespective of the type of APOL1-Vs expression. Similarly, the high throughput cell live-based assay showed a comparable increased action of the plasma membrane glycosphingolipid-glycohydrolases in living cells independent of the genetic APOL1 expression profile. Importantly, the most significative modification of the sphingolipid pattern induced by APOL1-Vs occurred in DRM resulted with a drastic reduction of radioactivity associated with sphingolipids. G1/G2-Vs present a decrease amount of globotriaosylceramide and globopentaosylceramide compared to G0. Additionally, ceramide at the DRM site and lactosylceramide in general, showed a greatest fall in G1/G2 in comparison with G0. Additionally, the levels of glucosylceramide decreased only in the DRM of human podocytes overexpressing G1/G2-Vs. These findings suggest that altered sphingolipidsprofiles may contribute to the deranged functionality of the plasma membrane in APOL1 risk milieu.

载脂蛋白 L1 (APOL1) 野生型 (G0) 在鞘脂、鞘糖脂、鞘磷脂和神经酰胺的代谢中起作用，它们构成了脂筏 (DRM) 的生物活性成分。我们询问 APOL1 变体 (APOL1-Vs) G1 和 G2 是否具有改变人类足细胞中鞘脂代谢的潜力。在惠普鞘脂图案过表达或者APOL1G0或APOL1-VS通过使用薄的单-和双维层色谱法，质谱分析法和代谢标记与[1-分析³H]鞘氨醇。HP G0 和 G1/G2-Vs 表现出可比的乳糖神经酰胺减少和球三糖神经酰胺含量的增加。对参与鞘糖脂分解代谢的主要糖水解酶活性的分析表明，受测酶的活性总体下降，而与 APOL1-Vs 表达的类型无关。类似地，基于高通量细胞活体的测定显示了与遗传 APOL1 表达谱无关的活细胞中质膜鞘糖脂-糖水解酶的可比增加的作用。重要的是，由 APOL1-Vs 诱导的鞘脂模式最显着的改变发生在 DRM 中，导致与鞘脂相关的放射性急剧下降。与 G0 相比，G1/G2-Vs 的 globotriaosylceramide 和 globopentaosylceramide 的量减少。此外，与 G0 相比，DRM 位点的神经酰胺和乳糖神经酰胺在 G1/G2 中的下降幅度最大。此外，葡萄糖神经酰胺的水平仅在过表达 G1/G2-Vs 的人足细胞的 DRM 中降低。这些发现表明，改变的鞘脂谱可能导致 APOL1 风险环境中质膜功能紊乱。