Mitochondria in gastric epithelial cells are the key targets for NSAIDs-induced injury and NGF cytoprotection.
Mitochondria in gastric epithelial cells are the key targets for NSAIDs-induced injury and NGF cytoprotection.
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胃上皮细胞中的线粒体是 NSAIDs 诱导的损伤和 NGF 细胞保护的关键靶标。
DOI：
10.1002/jcb.28445
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发表时间：
2019
影响因子：
4
通讯作者：
Tarnawski,AndrzejS
中科院分区：
文献类型：
--
作者：
Ahluwalia,Amrita;Jones,MichaelK;Hoa,Neil;Tarnawski,AndrzejS
胃上皮细胞是粘膜保护的重要组成部分，也是非甾体抗炎药（NSAIDs）诱导损伤的靶点。双氯芬酸（DFN）是应用最广泛的非甾体抗炎药之一；然而，即使短期使用也会引起胃腐蚀和溃疡。神经生长因子（NGF）不仅作用于神经元细胞，而且作用于内皮细胞；然而，其对胃上皮细胞的作用尚不清楚。本研究旨在确定NGF是否可以保护胃上皮细胞免受DFN诱导的损伤，并确定其潜在的分子机制，重点关注线粒体、生存素和胰岛素样生长因子1 （IGF - 1）。将培养的正常大鼠胃粘膜上皮细胞1 （RGM1）用磷酸盐缓冲盐水（PBS；对照）、NGF （100 ng/mL）和/或DFN（0.25‐1.00 mM）处理4小时。我们检查了：(1)共聚焦显微镜下的细胞损伤；(2) Calcein AM活细胞跟踪染料细胞死亡/存活；(3) MitoTracker在活细胞中的线粒体结构和膜电位功能；(4)免疫染色法检测NGF、其受体原肌球蛋白受体激酶A （TrkA）、survivin和IGF - 1的表达。DFN处理RGM1细胞4小时导致广泛的细胞损伤、线粒体解体、细胞活力降低（从对照组的94±3%降至0.5 mM DFN处理细胞的14±4%，P< 0.001），以及survivin和IGF - 1的表达。与PBS对照组相比，NGF治疗显著增加了41%和75%的survivin和IGF - 1表达。与单独使用DFN相比，在DFN治疗前使用NGF预处理可分别减少73%和82%的线粒体损伤和细胞死亡（均p < 0.001）。该研究还表明，在RGM1细胞的质膜和线粒体中存在高亲和力的TrkA受体，这表明NGF具有新的作用。
Gastric epithelial cells are important components of mucosal protection and targets of nonsteroidal anti‐inflammatory drugs (NSAIDs)‐induced injury. Diclofenac (DFN) is one of the most widely used NSAIDs; however, even its short‐term use can induce gastric erosions and ulcers. Nerve growth factor (NGF) has been reported to act not only on neuronal cells but also on endothelial cells; however, its action on gastric epithelial cells is unknown. This study was aimed to determine, whether NGF can protect gastric epithelial cells against DFN‐induced injury, and to determine the underlying molecular mechanisms with a focus on mitochondria, survivin, and insulin‐like growth factor 1 (IGF‐1). Cultured normal rat gastric mucosal epithelial cells 1 (RGM1) were treated with phosphate‐buffered saline (PBS; control), NGF (100 ng/mL) and/or DFN (0.25‐1.00 mM) for 4 hours. We examined: (1) cell injury by confocal microscopy; (2) cell death/survival using Calcein AM live cell tracking dye; (3) mitochondrial structure and membrane potential function using MitoTracker in live cells; and (4) expression of NGF, its receptor ‐ tropomyosin receptor kinase A (TrkA), survivin and IGF‐1 by immunostaining. DFN treatment of RGM1 cells for 4 hours caused extensive cell injury, mitochondrial disintegration, reduced cell viability (from 94 ± 3% in controls to 14 ± 4% in 0.5 mM DFN‐treated cells;P< 0.001), and expression of survivin and IGF‐1. NGF treatment significantly increased survivin and IGF‐1 expression by 41% and 75%, respectively versus PBS controls. Pretreatment with NGF before DFN treatment reduced mitochondrial damage and cell death by 73% and 82%, respectively versus treatment with DFN alone (allP< 0.001). This study also showed the presence of high‐affinity TrkA receptors in the plasma membrane and mitochondria of RGM1 cells indicating novel actions of NGF.