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Protective Effects of Neutrophil Gelatinase-Associated Lipocalin on Hypoxia/Reoxygenation Injury of HK-2 Cells.
MedLine Citation:
PMID:  22172816     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
BACKGROUND: Neutrophil gelatinase-associated lipocalin (NAGL) was first extracted from neutrophil granules. Our previous study showed that the expression of NGAL mRNA and protein can be induced by hypoxia/reoxygenation. This study was designed to investigate the relationship between NGAL and hypoxia/reoxygenation injury pathologies in HK-2 cells.
METHODS: The effect of NGAL on the proliferation of HK-2 cell lines was analyzed with a MTT colorimetric assay. Cell-cycle distribution and measurement of the percentage of apoptotic cells were performed by flow cytometry after stained with propidium iodide and annexin V-fluorescein isothiocyanate. The expression of genes for apoptotic proteins Bcl-2, Bax, and caspase-3 was analyzed with real-time reverse-transcription polymerase chain reaction (RT-PCR). The expression of NGAL mRNA and protein was analyzed with real-time RT-PCR or Western blot, respectively.
RESULTS: HK-2 cells were treated with hypoxia/reoxygenation. HK-2 cells exhibited an increase in the number of cells in the G0/G1 phase and a decrease in the number of cells in the S and G2/M phases. The proliferation index is decreased. When HK-2 cells were treated with 200 ng/mL recombinant NGAL and hypoxia/reoxygenation, there were no effects on cell-cycle distribution. The ratio of early apoptotic cells in the control and hypoxia/reoxygenation groups were 1.1% and 26.5%, respectively. After the addition of 200 ng/mL recombinant NGAL, the ratio of early apoptotic cells in the hypoxia/reoxygenation group dropped to 19.6%. The expression of Bax/Bcl-2 ratio and caspase-3 were significantly higher in the hypoxia/reoxygenation compared with the control group. After the addition of 200 ng/mL recombinant NGAL, the levels of Bax/Bcl-2 ratio and caspase-3 decreased significantly compared with the control group.
CONCLUSIONS: The action of NGAL against hypoxia/reoxygenation injury was due to inhibiting the apoptosis via inhibition of the expression of the genes of proapoptotic proteins Bax, Bcl-2, and caspase-3.
Authors:
L Y Cui; S Yang; J Zhang
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Transplantation proceedings     Volume:  43     ISSN:  1873-2623     ISO Abbreviation:  Transplant. Proc.     Publication Date:  2011 Dec 
Date Detail:
Created Date:  2011-12-16     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0243532     Medline TA:  Transplant Proc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3622-7     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Elsevier Inc. All rights reserved.
Affiliation:
Department of Laboratory Medicine, Peking University Third Hospital, Beijing, China.
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