Hyperosmolarity-Induced Apoptosis in Human Corneal Epithelial Cells Is Mediated by Cytochrome c and MAPK Pathways

细胞色素c 细胞凋亡 MAPK/ERK通路 激酶 分子生物学 生物 细胞生物学 蛋白激酶A 细胞外 化学 生物化学
作者
Lihui Luo,De‐Quan Li,Stephen C. Pflugfelder
出处
期刊:Cornea [Ovid Technologies (Wolters Kluwer)]
卷期号:26 (4): 452-460 被引量:158
标识
DOI:10.1097/ico.0b013e318030d259
摘要

Purpose: To study whether hyperosmolarity induces apoptosis in human corneal epithelial cells through cytochrome c-mediated death pathways and by activation of mitogen-activated protein kinases (MAPKs). Methods: Primary human corneal epithelial cells cultured in normal osmolar media (312 mOsM) were switched to hyperosmolar media (450, 500, and 550 mOsM) by adding 70, 90, and 120 mM NaCl, respectively, with or without the c-jun N-terminal kinase (JNK) inhibitor SB202190 or the extracellular-regulated kinase (ERK) inhibitor PD98059. Apoptosis was assessed by the ApopTag In Situ Oligo Ligation (ISOL) assay. Confocal microscopy was used to detect cytochrome c and active caspase-3. Total RNA was extracted and subjected to reverse transcriptase-polymerase chain reaction for apoptosis-associated genes. Western blots were performed on cell extracts for the apoptogenic molecules cytochrome c and Smac/DIABLO, and phospho-JNK and ERK. Results: ISOL-positive apoptotic cells significantly increased from 3.3 ± 1.6% in control medium to 11.4 ± 5.8%, 18.9 ± 4.8%, and 43.9 ± 8.8% in 70, 90, and 120 mM NaCl added media, respectively. The 90 mM NaCl high saline medium notably increased release of cytochrome c and Smac/DIABLO from mitochondria; activated caspase-3, JNK and ERK; stimulated mRNA expression of interleukin-1-converting enzyme and Bax; and reduced Bcl2 expression. SB202190 and PD98059 significantly suppressed hyperosmolarity-induced JNK/ERK activation and ISOL-positive cells. In addition, PD98059 inhibited the release of cytochrome c and Smac/DIABLO from mitochondria. Conclusions: These findings show that hyperosmolarity induces apoptosis of human corneal epithelial cells through a cytochrome c-mediated death pathway, which may be mediated by JNK and ERK MAPK signaling pathways.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
不配.应助思思采纳,获得30
1秒前
2秒前
ding应助呵呵呵悦采纳,获得10
2秒前
聪明的宛菡完成签到,获得积分10
2秒前
3秒前
野性的采枫完成签到,获得积分10
4秒前
lkl完成签到,获得积分10
4秒前
Ron完成签到,获得积分10
5秒前
wuwa完成签到,获得积分10
5秒前
5秒前
秋海棠完成签到,获得积分10
5秒前
114514完成签到 ,获得积分10
5秒前
6秒前
科研小白完成签到,获得积分10
6秒前
杪夏二八完成签到 ,获得积分10
7秒前
流浪发布了新的文献求助10
7秒前
大气乐儿完成签到,获得积分10
7秒前
111发布了新的文献求助10
8秒前
8秒前
CodeCraft应助崛起之邦采纳,获得30
8秒前
南桥完成签到 ,获得积分10
9秒前
zhuge完成签到,获得积分10
9秒前
陆陆完成签到,获得积分10
10秒前
空溟fever完成签到,获得积分10
10秒前
crrrr发布了新的文献求助10
10秒前
切奇莉亚发布了新的文献求助10
10秒前
ALMT完成签到,获得积分10
11秒前
zzr真真97完成签到,获得积分10
12秒前
bcsunny2022发布了新的文献求助10
13秒前
cdragon完成签到,获得积分10
13秒前
鳗鱼思卉完成签到,获得积分10
13秒前
东郭谷雪发布了新的文献求助10
14秒前
15秒前
小明完成签到,获得积分10
16秒前
Silence完成签到,获得积分10
16秒前
16秒前
16秒前
大地完成签到,获得积分10
17秒前
鲤鱼问雁完成签到,获得积分10
17秒前
高分求助中
Evolution 10000
Sustainability in Tides Chemistry 2800
The Young builders of New china : the visit of the delegation of the WFDY to the Chinese People's Republic 1000
юрские динозавры восточного забайкалья 800
English Wealden Fossils 700
Foreign Policy of the French Second Empire: A Bibliography 500
Chen Hansheng: China’s Last Romantic Revolutionary 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3147003
求助须知:如何正确求助?哪些是违规求助? 2798336
关于积分的说明 7827807
捐赠科研通 2454956
什么是DOI,文献DOI怎么找? 1306492
科研通“疑难数据库(出版商)”最低求助积分说明 627808
版权声明 601565