Institute of Biological Chemistry, Academia Sinica, No. 128, Academia Road Section 2, Nan-Kang, Taipei, 11529, Taiwan.
Department of Applied Chemistry, National Yang Ming Chiao Tung University, 1001, University Road, Eastern District, Hsinchu, 300093, Taiwan.
J Biomed Sci. 2024 Apr 29;31(1):44. doi: 10.1186/s12929-024-01031-8.
Helicobacter pylori, the main cause of various gastric diseases, infects approximately half of the human population. This pathogen is auxotrophic for cholesterol which it converts to various cholesteryl α-glucoside derivatives, including cholesteryl 6'-acyl α-glucoside (CAG). Since the related biosynthetic enzymes can be translocated to the host cells, the acyl chain of CAG likely comes from its precursor phosphatidylethanolamine (PE) in the host membranes. This work aims at examining how the acyl chain of CAG and PE inhibits the membrane functions, especially bacterial adhesion.
Eleven CAGs that differ in acyl chains were used to study the membrane properties of human gastric adenocarcinoma cells (AGS cells), including lipid rafts clustering (monitored by immunofluorescence with confocal microscopy) and lateral membrane fluidity (by the fluorescence recovery after photobleaching). Cell-based and mouse models were employed to study the degree of bacterial adhesion, the analyses of which were conducted by using flow cytometry and immunofluorescence staining, respectively. The lipidomes of H. pylori, AGS cells and H. pylori-AGS co-cultures were analyzed by Ultraperformance Liquid Chromatography-Tandem Mass Spectroscopy (UPLC-MS/MS) to examine the effect of PE(10:0), PE(18:0), PE(18:3), or PE(22:6) treatments.
CAG10:0, CAG18:3 and CAG22:6 were found to cause the most adverse effect on the bacterial adhesion. Further LC-MS analysis indicated that the treatment of PE(10:0) resulted in dual effects to inhibit the bacterial adhesion, including the generation of CAG10:0 and significant changes in the membrane compositions. The initial (1 h) lipidome changes involved in the incorporation of 10:0 acyl chains into dihydro- and phytosphingosine derivatives and ceramides. In contrast, after 16 h, glycerophospholipids displayed obvious increase in their very long chain fatty acids, monounsaturated and polyunsaturated fatty acids that are considered to enhance membrane fluidity.
The PE(10:0) treatment significantly reduced bacterial adhesion in both AGS cells and mouse models. Our approach of membrane remodeling has thus shown great promise as a new anti-H. pylori therapy.
幽门螺杆菌是各种胃病的主要病因，它感染了大约一半的人类。这种病原体是胆固醇营养缺陷型的，它将胆固醇转化为各种胆甾基α-葡糖苷衍生物，包括胆甾基 6′-酰基α-葡糖苷（CAG）。由于相关的生物合成酶可以转移到宿主细胞中，CAG 的酰基链很可能来自宿主膜中的其前体磷脂酰乙醇胺（PE）。本工作旨在研究 CAG 和 PE 的酰基链如何抑制膜功能，特别是细菌黏附。
使用 11 种不同酰基链的 CAG 研究人胃腺癌细胞（AGS 细胞）的膜性质，包括脂质筏聚集（通过共聚焦显微镜的免疫荧光监测）和侧向膜流动性（荧光恢复后光漂白）。细胞模型和小鼠模型用于研究细菌黏附程度，分别通过流式细胞术和免疫荧光染色进行分析。通过超高效液相色谱-串联质谱（UPLC-MS/MS）分析 H. pylori、AGS 细胞和 H. pylori-AGS 共培养物的脂质组，以检查 PE(10:0)、PE(18:0)、PE(18:3)或 PE(22:6)处理的影响。
发现 CAG10:0、CAG18:3 和 CAG22:6 对细菌黏附的影响最不利。进一步的 LC-MS 分析表明，PE(10:0)处理具有双重抑制细菌黏附的作用，包括生成 CAG10:0 和显著改变膜组成。最初（1 小时）脂质组变化涉及将 10:0 酰基链掺入二氢和植物鞘氨醇衍生物和神经酰胺中。相比之下，16 小时后，甘油磷脂显示出其非常长链脂肪酸、单不饱和和多不饱和脂肪酸的明显增加，这些脂肪酸被认为可以增强膜流动性。
PE(10:0)处理在 AGS 细胞和小鼠模型中均显著降低了细菌黏附。我们的膜重塑方法因此显示出作为一种新的抗 H. pylori 治疗方法的巨大潜力。
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