Lipopolysaccharide effects on the proliferation of NRK52E cells via alternations in gap-junction function

Ziqing Hei, Ailan Zhang, Jing Wei, Xiaoliang Gan, Yanling Wang, Gangjian Luo, Xiaoyun Li

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)67-72
Number of pages6
JournalJournal of Trauma and Acute Care Surgery
Volume73
Issue number1
DOIs
StatePublished - Jul 1 2012

Keywords

  • Lipopolysaccharide; gap junction; connexin; renal injury

ASJC Scopus subject areas

  • Surgery
  • Critical Care and Intensive Care Medicine

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