The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells

To determine the effect of hyperosmolarity on cell survival/apoptosis of conjunctival epithelial cells and evaluate the possible role of IL6, Wong-Kilbourne derivative of Chang conjunctival cell line (WKD) was used in this study. Confluent cells were incubated under different osmolarity (290 mOsm an...

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Main Authors: Hee-Jung Ju, Yong-Soo Byun, Jee-Won Mok, Choun-Ki Joo
Format: Article
Language:English
Published: Wiley 2016-01-01
Series:Journal of Ophthalmology
Online Access:http://dx.doi.org/10.1155/2016/8350134
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author Hee-Jung Ju
Yong-Soo Byun
Jee-Won Mok
Choun-Ki Joo
author_facet Hee-Jung Ju
Yong-Soo Byun
Jee-Won Mok
Choun-Ki Joo
author_sort Hee-Jung Ju
collection DOAJ
description To determine the effect of hyperosmolarity on cell survival/apoptosis of conjunctival epithelial cells and evaluate the possible role of IL6, Wong-Kilbourne derivative of Chang conjunctival cell line (WKD) was used in this study. Confluent cells were incubated under different osmolarity (290 mOsm and 500 mOsm) with or without neutralizing IL6 antibody (50 ng/mL). The expression of IL6 level was measured in the supernatant of each conditioned medium. Cell viability/apoptosis assay was performed using Annexin V/Propidium Iodide (PI) and Cell Counting Kit-8 (CCK-8). Western blot was conducted to measure the abundance of apoptotic markers and IL6 related downstream signaling pathway. The concentration of IL6 showed time-dependent increase in cells treated with 500 mOsm. Although apoptosis of WKD cell is increased in treated 500 mOsm for 24 h, apoptosis reduced in WKD cell treated 500 mOsm with anti-IL6 for 24 h. Anti-IL6 inhibited the activation of JAK-STAT signaling pathway, which was induced by hyperosmolarity. Hyperosmolar condition induced apoptosis in conjunctival epithelial cells, along with increase of IL6 production. IL6 neutralizing antibody inhibited apoptosis and JAK-STAT signaling in hyperosmolar condition. These findings suggested that IL6 may be involved in apoptotic change and in hyperosmolarity.
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spelling doaj-art-a32750ea225c4d6b8ca4a6cff8b71bb62025-02-03T01:00:56ZengWileyJournal of Ophthalmology2090-004X2090-00582016-01-01201610.1155/2016/83501348350134The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial CellsHee-Jung Ju0Yong-Soo Byun1Jee-Won Mok2Choun-Ki Joo3Catholic Institute for Visual Science, Seoul St. Mary’s Hospital, Seoul 06591, Republic of KoreaCatholic Institute for Visual Science, Seoul St. Mary’s Hospital, Seoul 06591, Republic of KoreaCatholic Institute for Visual Science, Seoul St. Mary’s Hospital, Seoul 06591, Republic of KoreaCatholic Institute for Visual Science, Seoul St. Mary’s Hospital, Seoul 06591, Republic of KoreaTo determine the effect of hyperosmolarity on cell survival/apoptosis of conjunctival epithelial cells and evaluate the possible role of IL6, Wong-Kilbourne derivative of Chang conjunctival cell line (WKD) was used in this study. Confluent cells were incubated under different osmolarity (290 mOsm and 500 mOsm) with or without neutralizing IL6 antibody (50 ng/mL). The expression of IL6 level was measured in the supernatant of each conditioned medium. Cell viability/apoptosis assay was performed using Annexin V/Propidium Iodide (PI) and Cell Counting Kit-8 (CCK-8). Western blot was conducted to measure the abundance of apoptotic markers and IL6 related downstream signaling pathway. The concentration of IL6 showed time-dependent increase in cells treated with 500 mOsm. Although apoptosis of WKD cell is increased in treated 500 mOsm for 24 h, apoptosis reduced in WKD cell treated 500 mOsm with anti-IL6 for 24 h. Anti-IL6 inhibited the activation of JAK-STAT signaling pathway, which was induced by hyperosmolarity. Hyperosmolar condition induced apoptosis in conjunctival epithelial cells, along with increase of IL6 production. IL6 neutralizing antibody inhibited apoptosis and JAK-STAT signaling in hyperosmolar condition. These findings suggested that IL6 may be involved in apoptotic change and in hyperosmolarity.http://dx.doi.org/10.1155/2016/8350134
spellingShingle Hee-Jung Ju
Yong-Soo Byun
Jee-Won Mok
Choun-Ki Joo
The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells
Journal of Ophthalmology
title The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells
title_full The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells
title_fullStr The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells
title_full_unstemmed The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells
title_short The Blockade of IL6 Counterparts the Osmolar Stress-Induced Apoptosis in Human Conjunctival Epithelial Cells
title_sort blockade of il6 counterparts the osmolar stress induced apoptosis in human conjunctival epithelial cells
url http://dx.doi.org/10.1155/2016/8350134
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