Slomiany A, Slomiany B L
Research Center University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA.
J Physiol Pharmacol. 2003 Dec;54(4):533-51.
Nuclear transcriptome initiates specific proteome that facilitates metabolic events culminating in restitution of cell components and reproduction of the discrete cellular function, but the magnitude of various genes induction and following proteomic, lipidomic, and glycomic processes provide distinctness to the final product and its function. In homeostasis, the challenged cell responds to stimuli in defined and predictable mode but in the disease such as ulcerative erosions the ablation of cell survival signals and cell apoptosis is enhanced. Therefore, to uncover the discreteness and dissimilarity of the pathological processes induced by elicobacter pylori (H. pylori) lipopolysaccharide (LPS), not only measurement of the genomic events is crucial, but a complete cycle of events reproducing the cell specific proteins, lipids, and cell-specific environment created in situ require thorough investigation.
An impact of H. pylori LPS-induced processes on posttranslational lipidomic activity in endoplasmic reticulum (ER), Golgi and apical membrane was evaluated in the in vitro paradigm assembled with components of the rat gastric mucosal epithelial cells.
In ER, the signals commanding synthesis of biomembrane in the presence of control, the LPS-derived or LPS-admixed cytosol was identical. The assembled vesicles contained the same amount of apoprotein and had the same lipid composition. Their biomembrane contained the same amount of sphingolipids in form of ceramide, which is determining factor of the ER-transport vesicle completion. The transport of apoprotein in ER vesicles to Golgi was also not changed. In Golgi, LPS-derived cytosol affected two distinct and concurrent with assembly of Golgi transport vesicles processes. The LPS-derived cytosol affected formation of Golgi transport vesicles destined to apical membrane and the incorporation (fusion) of Golgi vesicles with apical epithelial membrane. The LPS-derived cytosol decreased the production of Golgi vesicles by 15% and their fusion with the apical epithelial membrane by 83%. In contrast with wortmannin, the LPS-derived cytosol had no impact on Golgi transport vesicles association with the epithelial membrane.
We concluded that LPS interferes with MAPK-dependent activation of cytosolic PLA(2) since MAPKs immunoprecipitate added to the LPS-cytosol restored activation of cytosolic PLA(2)-specific fusion of the Golgi transport vesicles with apical mucosal cell membrane. On the other hand, wortmannin that inhibited the association of Golgi transport vesicles with apical membrane, interferes with cytosolic activity that controls association of PI3K-containing Golgi vesicles with the apical membrane. Together, our studies present evidence that allow to conclude that LPS affects MAPK-specific phosphorylation and PLA(2)-assisted membranes' fusion, whereas wortmannin affects association of PI3K- and PI3P-containing Golgi-derived transport vesicles with the membrane. In the final outcome, both actions result in a diminished or inhibited restitution of apical membrane.
核转录组启动特定的蛋白质组，促进代谢事件，最终恢复细胞成分并重现离散的细胞功能，但各种基因诱导的程度以及随后的蛋白质组学、脂质组学和糖组学过程赋予最终产物及其功能独特性。在稳态下，受挑战的细胞以明确且可预测的方式对刺激作出反应，但在诸如溃疡性糜烂等疾病中，细胞存活信号的缺失和细胞凋亡会增强。因此，为了揭示幽门螺杆菌（H. pylori）脂多糖（LPS）诱导的病理过程的离散性和差异性，不仅基因组事件的测量至关重要，而且完整的事件循环，即重现原位产生的细胞特异性蛋白质、脂质和细胞特异性环境，需要深入研究。
在由大鼠胃黏膜上皮细胞成分组装而成的体外模型中，评估幽门螺杆菌LPS诱导的过程对内质网（ER）、高尔基体和顶端膜中翻译后脂质组学活性的影响。
在内质网中，在对照、LPS衍生的或LPS混合的胞质溶胶存在下，指挥生物膜合成的信号是相同的。组装的囊泡含有相同量的载脂蛋白，并且具有相同的脂质组成。它们的生物膜含有相同量的神经酰胺形式的鞘脂，这是内质网运输囊泡完成的决定因素。内质网囊泡中载脂蛋白向高尔基体的运输也未改变。在高尔基体中，LPS衍生的胞质溶胶影响两个不同且与高尔基体运输囊泡组装同时发生的过程。LPS衍生的胞质溶胶影响 destined to apical membrane高尔基体运输囊泡的形成以及高尔基体囊泡与顶端上皮膜的融合。LPS衍生的胞质溶胶使高尔基体囊泡的产生减少了15%，其与顶端上皮膜的融合减少了83%。与渥曼青霉素不同，LPS衍生的胞质溶胶对高尔基体运输囊泡与上皮膜的结合没有影响。
我们得出结论，LPS干扰了丝裂原活化蛋白激酶（MAPK）依赖性的胞质磷脂酶A2（cytosolic PLA(2)）的激活，因为添加到LPS - 胞质溶胶中的MAPKs免疫沉淀物恢复了胞质溶胶PLA(2) - 高尔基体运输囊泡与顶端黏膜细胞膜特异性融合的激活。另一方面，抑制高尔基体运输囊泡与顶端膜结合的渥曼青霉素，干扰了控制含PI3K的高尔基体囊泡与顶端膜结合的胞质活性。总之，我们的研究提供的证据表明，可以得出结论，LPS影响MAPK特异性磷酸化和PLA(2)辅助的膜融合，而渥曼青霉素影响含PI3K和PI3P的高尔基体衍生运输囊泡与膜的结合。最终结果是，这两种作用都导致顶端膜的恢复减少或受到抑制。
