Ganglioside GM1 is a glycosphingolipid found on mammalian cell membranes, and it is involved in ischemic encephalopathy, spinal cord injury and neurodegenerative diseases. Fatty acids, as a structure module of GM1, have been reported to affect its physiological function and neurite growth activity. Due to the limitation of preparation methods, the function of GM1 derivatives containing different fatty acids in nerve cells has not been systematically studied. To discover novel GM1 derivatives as nerve growth-promoting agents, we developed an efficient SA_SCDase enzymatic synthetic system of GM1 derivatives, yielding twenty GM1 derivatives with unsaturated fatty acid chains in high total yields (16–67%). Subsequently, the neurite outgrowth activities of GM1 derivatives were assessed on Neuro2a Cells. Among all the GM1 derivatives, GM1 (d18:1/C16:1) induced demonstrably neurite outgrowth activity. The subsequent RNA-sequencing (RNA-seq) and Western blot analysis was then performed and indicated that the mechanism of nerve cells growth involved cholesterol biosynthesis regulation by up-regulating SREBP2 expression or ERBB4 phosphorylation to activate the PI3K-mTOR pathway.

神经节苷脂 GM1 是一种存在于哺乳动物细胞膜上的鞘糖脂，它与缺血性脑病、脊髓损伤和神经退行性疾病有关。据报道，脂肪酸作为 GM1 的结构模块，会影响其生理功能和神经突生长活性。由于制备方法的限制，含有不同脂肪酸的GM1衍生物在神经细胞中的作用尚未得到系统研究。为了发现新的 GM1 衍生物作为神经生长促进剂，我们开发了一种高效的 SA_SCDase 酶促合成 GM1 衍生物系统，以高总产率 (16-67%) 产生了 20 种具有不饱和脂肪酸链的 GM1 衍生物。随后，在 Neuro2a 细胞上评估了 GM1 衍生物的神经突生长活性。在所有 GM1 衍生物中，GM1 (d18:1/C16: 1) 诱导明显的神经突生长活动。随后进行了 RNA 测序 (RNA-seq) 和蛋白质印迹分析，表明神经细胞生长的机制涉及通过上调 SREBP2 表达或 ERBB4 磷酸化来激活 PI3K-mTOR 通路来调节胆固醇生物合成。