Institute of Life Science, Swansea University Medical School, Swansea University, Swansea, Wales, United Kingdom.
PLoS One. 2020 Mar 12;15(3):e0219275. doi: 10.1371/journal.pone.0219275. eCollection 2020.
Pathogenic bacteria often damage tissues by secreting toxins that form pores in cell membranes, and the most common pore-forming toxins are cholesterol-dependent cytolysins. During bacterial infections, glutamine becomes a conditionally essential amino acid, and glutamine is an important nutrient for immune cells. However, the role of glutamine in protecting tissue cells against pore-forming toxins is unclear. Here we tested the hypothesis that glutamine supports the protection of tissue cells against the damage caused by cholesterol-dependent cytolysins. Stromal and epithelial cells were sensitive to damage by the cholesterol-dependent cytolysins, pyolysin and streptolysin O, as determined by leakage of potassium and lactate dehydrogenase from cells, and reduced cell viability. However, glutamine deprivation increased the leakage of lactate dehydrogenase and reduced the viability of cells challenged with cholesterol-dependent cytolysins. Without glutamine, stromal cells challenged with pyolysin leaked lactate dehydrogenase (control vs. pyolysin, 2.6 ± 0.6 vs. 34.4 ± 4.5 AU, n = 12), which was more than three-fold the leakage from cells supplied with 2 mM glutamine (control vs. pyolysin, 2.2 ± 0.3 vs. 9.4 ± 1.0 AU). Glutamine cytoprotection did not depend on glutaminolysis, replenishing the Krebs cycle via succinate, changes in cellular cholesterol, or regulators of cell metabolism (AMPK and mTOR). In conclusion, although the mechanism remains elusive, we found that glutamine supports the protection of tissue cells against the damage caused by cholesterol-dependent cytolysins from pathogenic bacteria.
致病细菌通常通过分泌在细胞膜中形成孔的毒素来损伤组织,而最常见的孔形成毒素是胆固醇依赖性细胞溶素。在细菌感染期间,谷氨酰胺成为条件必需氨基酸,并且是免疫细胞的重要营养物质。然而,谷氨酰胺在保护组织细胞免受孔形成毒素损害中的作用尚不清楚。在这里,我们检验了这样一个假设,即谷氨酰胺支持组织细胞抵抗胆固醇依赖性细胞溶素引起的损伤。基质细胞和上皮细胞对胆固醇依赖性细胞溶素(如 pyolysin 和 streptolysin O)敏感,这表现在细胞内钾和乳酸脱氢酶的漏出以及细胞活力降低。然而,谷氨酰胺剥夺增加了乳酸脱氢酶的漏出,并降低了受到胆固醇依赖性细胞溶素挑战的细胞活力。没有谷氨酰胺,基质细胞受到 pyolysin 的攻击会漏出乳酸脱氢酶(对照与 pyolysin,2.6 ± 0.6 对 34.4 ± 4.5 AU,n = 12),是用 2 mM 谷氨酰胺供应的细胞漏出的三倍多(对照与 pyolysin,2.2 ± 0.3 对 9.4 ± 1.0 AU)。谷氨酰胺细胞保护作用不依赖于谷氨酰胺分解代谢,通过琥珀酸补充克雷布斯循环,细胞胆固醇变化或细胞代谢调节剂(AMPK 和 mTOR)都不依赖于谷氨酰胺分解代谢。总之,尽管机制尚不清楚,但我们发现谷氨酰胺支持组织细胞抵抗致病性细菌产生的胆固醇依赖性细胞溶素引起的损伤。