Document Detail

Lipopolysaccharide effects on the proliferation of NRK52E cells via alternations in gap-junction function.
MedLine Citation:
PMID:  22743374     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Gap junctions regulate proper kidney function by facilitating intercellular communication, vascular conduction, and tubular purinergic signaling. However, no clear relationship has been described between gap-junction function and acute kidney injury induced by the endotoxin lipopolysaccharide (LPS).
METHODS: Normal rat kidney epithelial cells (NRK52E cells) were seeded at high and low densities to promote or impede gap-junction formation, respectively, and establish distinctive levels of intercellular communication in culture. Cells were then challenged with LPS at various concentrations (10-1,000 ng/mL). LPS-induced formation and function of gap junctions were assessed by measuring changes in cell proliferation and colony-forming rates, fluorescent dye transmission to adjacent cells, expression levels of connexin43, and repositioning of confluent cells in response to the gap junction inhibitor oleamide or agonist retinoic acid.
RESULTS: The cell proliferation rate and colony-forming rate of high- and low-density NRK52E cells were decreased upon LPS challenge, in a dose-dependent manner. The colony-forming rate of confluent high-density cells was significantly lower than that of low-density cells. Oleamide treatment raised the LPS-induced colony-forming rate of high-density cells, whereas retinoic acid decreased the rate. Neither oleamide nor retinoic acid significantly affected the LPS-induced colony-forming rate of low-density cells. Fluorescence transmission of high-density cells was reduced by LPS challenge, in a dose-dependent manner, but inclusion of retinoic acid increased the LPS-induced transmission of fluorescence. LPS challenge of either high- or low-density NRK52E cells resulted in down-regulated connexin43 expression.
CONCLUSION: Gap-junction function plays an important role in concentration-dependent cytotoxic effect of LPS on normal rat kidney cells in vitro.
Ziqing Hei; Ailan Zhang; Jing Wei; Xiaoliang Gan; Yanling Wang; Gangjian Luo; Xiaoyun Li
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The journal of trauma and acute care surgery     Volume:  73     ISSN:  2163-0763     ISO Abbreviation:  J Trauma Acute Care Surg     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-06-29     Completed Date:  2012-09-17     Revised Date:  2013-09-25    
Medline Journal Info:
Nlm Unique ID:  101570622     Medline TA:  J Trauma Acute Care Surg     Country:  United States    
Other Details:
Languages:  eng     Pagination:  67-72     Citation Subset:  AIM; IM    
Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
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MeSH Terms
Blotting, Western
Cell Line
Cell Proliferation / drug effects*
Connexin 43 / physiology
Dose-Response Relationship, Drug
Endotoxins / pharmacology
Gap Junctions / drug effects,  physiology*
Kidney / cytology
Lipopolysaccharides / pharmacology*
Oleic Acids / pharmacology
Stem Cells / drug effects,  physiology
Tretinoin / pharmacology
Reg. No./Substance:
0/Connexin 43; 0/Endotoxins; 0/Lipopolysaccharides; 0/Oleic Acids; 302-79-4/Tretinoin; 67924-63-4/endotoxin, Escherichia coli; 7L25QK8BWO/oleylamide

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