Cargando…

17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway

BACKGROUND: Tumor necrosis factor-α (TNF-α) has been widely known to induce degeneration of nucleus pulposus cells (NPCs). 17β-estradiol (17β-E2) has been broadly proven for its function of suppressing cell apoptosis. The aim of this study is to explore whether 17β-E2 protects apoptosis of human NPC...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Tao, Yang, Si-Dong, Liu, Sen, Wang, Hui, Liu, Huan, Ding, Wen-Yuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Scientific Literature, Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5115218/
https://www.ncbi.nlm.nih.gov/pubmed/27847386
http://dx.doi.org/10.12659/MSM.900310
_version_ 1782468487426342912
author Wang, Tao
Yang, Si-Dong
Liu, Sen
Wang, Hui
Liu, Huan
Ding, Wen-Yuan
author_facet Wang, Tao
Yang, Si-Dong
Liu, Sen
Wang, Hui
Liu, Huan
Ding, Wen-Yuan
author_sort Wang, Tao
collection PubMed
description BACKGROUND: Tumor necrosis factor-α (TNF-α) has been widely known to induce degeneration of nucleus pulposus cells (NPCs). 17β-estradiol (17β-E2) has been broadly proven for its function of suppressing cell apoptosis. The aim of this study is to explore whether 17β-E2 protects apoptosis of human NPCs induced by TNF-α via the PI3K/AKT pathway. MATERIAL/METHODS: NPCs were divided into four groups: control, TNF-α (100 ng/mL), TNF-α (100 ng/mL) with pretreated 17β-E2 (10 um/L), TNF-α (100 ng/mL) with pretreated 17β-E2 (10 um/L) and MK2206 (10 um/L, inhibitor of the PI3K/AKT pathway). Flow cytometry was used to measure the apoptotic incidence. Inverted phase-contrast microscopy was used to accomplish the morphological observation for apoptosis of treated cells. Additionally, Cell Counting Kit 8 (CCK-8) assay was used to detected cell proliferation. Western blot and quantitative real-time PCR (qRT-PCR) were applied to explore the expression of pro-caspase-3, caspase-3/p17, cleaved PARP, PARP, Akt, and phospho-Akt (p-Akt). RESULTS: First, inverted phase-contrast microscopy, CCK-8, and flow cytometry showed that TNF-α induced marked apoptosis, which was abolished by 17β-E2. Furthermore, Western blot and qRT-PCR showed that 17β-E2 protects TNF-α which can induced apoptosis by upregulating p-Akt, whereas Akt was essentially constant. Our data revealed that p-Akt expression peaked at 24 hours in a time-dependent manner (0–48 hours) after treating with TNF-α; and the p-Akt expression generally increased in a time-dependent manner (0–48 hours) after treating with TNF-α and 17β-E2. CONCLUSIONS: 17β-E2 is shown to protect NPCs against TNF-α induced apoptosis by upregulating p-Akt in the PI3K/AKT pathway. 17β-E2 generally increases expression of p-Akt.
format Online
Article
Text
id pubmed-5115218
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher International Scientific Literature, Inc.
record_format MEDLINE/PubMed
spelling pubmed-51152182016-11-23 17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway Wang, Tao Yang, Si-Dong Liu, Sen Wang, Hui Liu, Huan Ding, Wen-Yuan Med Sci Monit Lab/In Vitro Research BACKGROUND: Tumor necrosis factor-α (TNF-α) has been widely known to induce degeneration of nucleus pulposus cells (NPCs). 17β-estradiol (17β-E2) has been broadly proven for its function of suppressing cell apoptosis. The aim of this study is to explore whether 17β-E2 protects apoptosis of human NPCs induced by TNF-α via the PI3K/AKT pathway. MATERIAL/METHODS: NPCs were divided into four groups: control, TNF-α (100 ng/mL), TNF-α (100 ng/mL) with pretreated 17β-E2 (10 um/L), TNF-α (100 ng/mL) with pretreated 17β-E2 (10 um/L) and MK2206 (10 um/L, inhibitor of the PI3K/AKT pathway). Flow cytometry was used to measure the apoptotic incidence. Inverted phase-contrast microscopy was used to accomplish the morphological observation for apoptosis of treated cells. Additionally, Cell Counting Kit 8 (CCK-8) assay was used to detected cell proliferation. Western blot and quantitative real-time PCR (qRT-PCR) were applied to explore the expression of pro-caspase-3, caspase-3/p17, cleaved PARP, PARP, Akt, and phospho-Akt (p-Akt). RESULTS: First, inverted phase-contrast microscopy, CCK-8, and flow cytometry showed that TNF-α induced marked apoptosis, which was abolished by 17β-E2. Furthermore, Western blot and qRT-PCR showed that 17β-E2 protects TNF-α which can induced apoptosis by upregulating p-Akt, whereas Akt was essentially constant. Our data revealed that p-Akt expression peaked at 24 hours in a time-dependent manner (0–48 hours) after treating with TNF-α; and the p-Akt expression generally increased in a time-dependent manner (0–48 hours) after treating with TNF-α and 17β-E2. CONCLUSIONS: 17β-E2 is shown to protect NPCs against TNF-α induced apoptosis by upregulating p-Akt in the PI3K/AKT pathway. 17β-E2 generally increases expression of p-Akt. International Scientific Literature, Inc. 2016-11-12 /pmc/articles/PMC5115218/ /pubmed/27847386 http://dx.doi.org/10.12659/MSM.900310 Text en © Med Sci Monit, 2016 This work is licensed under Creative Common Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
spellingShingle Lab/In Vitro Research
Wang, Tao
Yang, Si-Dong
Liu, Sen
Wang, Hui
Liu, Huan
Ding, Wen-Yuan
17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
title 17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
title_full 17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
title_fullStr 17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
title_full_unstemmed 17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
title_short 17β-Estradiol Inhibites Tumor Necrosis Factor-α Induced Apoptosis of Human Nucleus Pulposus Cells via the PI3K/Akt Pathway
title_sort 17β-estradiol inhibites tumor necrosis factor-α induced apoptosis of human nucleus pulposus cells via the pi3k/akt pathway
topic Lab/In Vitro Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5115218/
https://www.ncbi.nlm.nih.gov/pubmed/27847386
http://dx.doi.org/10.12659/MSM.900310
work_keys_str_mv AT wangtao 17bestradiolinhibitestumornecrosisfactorainducedapoptosisofhumannucleuspulposuscellsviathepi3kaktpathway
AT yangsidong 17bestradiolinhibitestumornecrosisfactorainducedapoptosisofhumannucleuspulposuscellsviathepi3kaktpathway
AT liusen 17bestradiolinhibitestumornecrosisfactorainducedapoptosisofhumannucleuspulposuscellsviathepi3kaktpathway
AT wanghui 17bestradiolinhibitestumornecrosisfactorainducedapoptosisofhumannucleuspulposuscellsviathepi3kaktpathway
AT liuhuan 17bestradiolinhibitestumornecrosisfactorainducedapoptosisofhumannucleuspulposuscellsviathepi3kaktpathway
AT dingwenyuan 17bestradiolinhibitestumornecrosisfactorainducedapoptosisofhumannucleuspulposuscellsviathepi3kaktpathway