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A new in vitro model of sepsis‐induced renal epithelial cell injury
Author(s) -
Pathak Elina,
Mayeux Philip R
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.754.2
Subject(s) - sepsis , lactate dehydrogenase , kidney , acute kidney injury , in vitro , medicine , nitric oxide , pharmacology , endocrinology , chemistry , immunology , biochemistry , enzyme
Previous studies by our laboratory have shown that sepsis produces a rapid inflammatory response in the kidney that leads to tubular injury. There are no in vitro models that adequately replicate the tubular microenvironment during sepsis. To address this need, we have established a new model of sepsis‐induced renal epithelial injury using serum from septic mice and a mouse renal epithelial cell line. mIMCD‐3 cells (epithelial cells from mouse inner medullary collecting duct) were treated with media containing 5% serum collected from mice at 4h that have undergone sham surgery (Sham) or cecal ligation and puncture (CLP) to induce sepsis. We chose 4h CLP serum because cytokines are elevated at this time. Real‐time PCR analysis showed a time‐dependent induction in iNOS mRNA in cells treated with CLP compared to Sham (1.4 ± 0.5 fold at 6h and 5.3 ± 0.7 fold at 18h; P < .05 compared to Sham). This was accompanied by a two‐fold increase in nitric oxide generation at 6h as measured by total nitrate/nitrite levels (P < .05 compared to Sham). We also measured lactate dehydrogenase release (LDH) to assess toxicity and found a two‐fold increase in LDH at 18h in CLP‐treated cells (P < .05 compared to Sham). These data suggest that exposure of tubular epithelial cells to serum from septic mice is a unique, highly relevant in vitro model with which to study the mechanism of sepsis‐induced kidney injury. Supported by NIH DK075991.