Department of Gastroenterology, JCHO Hoshigaoka Medical Center, 4-8-1, Hoshigaoka, Hirakata, 573-8511, Japan.
Department of Internal Medicine, Higashisumiyoshi Morimoto Hospital, 3-2-66, Takaai, Higashi-Sumiyoshi-Ku, Osaka, 546-0014, Japan.
J Pharmacol Sci. 2023 Apr;151(4):163-170. doi: 10.1016/j.jphs.2023.02.001. Epub 2023 Feb 7.
Mast cell-derived tryptase causes neuronal elongation/sensitization leading to visceral hypersensitivity. However, effects of tryptase on enteric glial cells (EGCs) and subsequent interaction between EGCs and neurons remain unknown.
We evaluated proteins and mRNA expressions in EGC (CRL-2690, ATCC) after tryptase stimulation: nerve growth factor (NGF), netrin-1, and glial cell-derived neurotrophic factor (GDNF). We examined morphological changes in neurons (PC12 cells, CRL-1721.1) by co-incubation with the conditioned medium of EGCs after tryptase stimulation.
EGC was activated by tryptase, and proliferated (by 1.8-fold) with cytoplasmic expansion and process elongation. Intercellular connections of EGC were more complexed. Tryptase induced mRNA expression (2.5-fold) and protein expression of NGF. Netrin-1 (3-fold) and GDNF (3-fold) mRNA expressions were increased at 30 min. Increase in netrin-1 continued until 6 h, whereas the latter decreased by 3 h. The conditioned medium of EGC after tryptase stimulation expanded neuronal cytoplasm (round or ramified shapes) and neurite outgrowth with elongation of cytoskeletal filaments in time-dependent and dose-dependent manners. These changes were similar to those after NGF stimulation. Growth cone proteins of neurons were also increased by the conditioned medium.
EGC activated by tryptase changes neuronal morphology (process elongation and cytoplasm expansion) possibly via the stimuli-associated mediators.
肥大细胞衍生的类胰蛋白酶导致神经元伸长/致敏，从而引起内脏敏感性。然而，类胰蛋白酶对肠神经胶质细胞（EGC）的影响以及随后 EGC 与神经元之间的相互作用仍不清楚。
我们评估了类胰蛋白酶刺激后 EGC（CRL-2690，ATCC）中的蛋白质和 mRNA 表达：神经生长因子（NGF）、轴突导向因子 1（netrin-1）和胶质细胞源性神经营养因子（GDNF）。我们通过与类胰蛋白酶刺激后 EGC 的条件培养基共孵育，观察神经元（PC12 细胞，CRL-1721.1）的形态变化。
类胰蛋白酶激活 EGC，引起细胞增殖（增加 1.8 倍），伴有细胞质扩张和突起伸长。EGC 之间的细胞间连接更加复杂。类胰蛋白酶诱导 NGF 的 mRNA 表达（增加 2.5 倍）和蛋白表达。Netrin-1（增加 3 倍）和 GDNF（增加 3 倍）的 mRNA 表达在 30 分钟时增加。Netrin-1 的增加持续到 6 小时，而后者在 3 小时时减少。类胰蛋白酶刺激后的 EGC 条件培养基以时间和剂量依赖的方式使神经元细胞质（圆形或分支形状）扩张和突起生长，细胞骨架丝伸长。这些变化与 NGF 刺激后的变化相似。神经元的生长锥蛋白也被条件培养基增加。
类胰蛋白酶激活的 EGC 通过与刺激相关的介质改变神经元形态（突起伸长和细胞质扩张）。
