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Interleukin-10 Down-Regulates Cathepsin B Expression in Fetal Rat Alveolar Type II Cells Exposed to Hyperoxia
Yonsei Medical Journal ; : 445-452, 2013.
Article in English | WPRIM | ID: wpr-89563
ABSTRACT

PURPOSE:

Hyperoxia has the chief biological effect of cell death. We have previously reported that cathepsin B (CB) is related to fetal alveolar type II cell (FATIIC) death and pretreatment of recombinant IL-10 (rIL-10) attenuates type II cell death during 65%-hyperoixa. In this study, we investigated what kinds of changes of CB expression are induced in FATIICs at different concentrations of hyperoxia (65%- and 85%-hyperoxia) and whether pretreatment with rIL-10 reduces the expression of CB in FATIICs during hyperoxia. MATERIALS AND

METHODS:

Isolated embryonic day 19 fetal rat alveolar type II cells were cultured and exposed to 65%- and 85%-hyperoxia for 12 h and 24 h. Cells in room air were used as controls. Cytotoxicity was assessed by lactate dehydrogenase (LDH) released into the supernatant. Expression of CB was analyzed by fluorescence-based assay upon cell lysis and western blotting, and LDH-release was re-analyzed after preincubation of cathepsin B-inhibitor (CBI). IL-10 production was analyzed by ELISA, and LDH-release was re-assessed after preincubation with rIL-10 and CB expression was re-analyzed by western blotting and real-time PCR.

RESULTS:

LDH-release and CB expression in FATIICs were enhanced significantly in an oxygen-concentration-dependent manner during hyperoxia, whereas caspase-3 was not activated. Preincubation of FATIICs with CBI significantly reduced LDH-release during hyperoxia. IL-10-release decreased in an oxygen-concentration-dependent fashion, and preincubation of the cells with rIL-10 significantly reduced cellular necrosis and expression of CB in FATIICs which were exposed to 65%- and 85%-hyperoxia.

CONCLUSION:

Our study suggests that CB is enhanced in an oxygen-concentration-dependent manner, and IL-10 has an inhibitory effect on CB expression in FATIICs during hyperoxia.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Oxygen / Cathepsin B / Down-Regulation / Gene Expression Regulation / Interleukin-10 / Hyperoxia / L-Lactate Dehydrogenase / Necrosis Limits: Animals Language: English Journal: Yonsei Medical Journal Year: 2013 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Oxygen / Cathepsin B / Down-Regulation / Gene Expression Regulation / Interleukin-10 / Hyperoxia / L-Lactate Dehydrogenase / Necrosis Limits: Animals Language: English Journal: Yonsei Medical Journal Year: 2013 Type: Article