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Intralipid Protects against Hypoxia/Reoxygenation Injury in Human Coronary Artery Endothelial Cells and Cardiomyocytes
Author(s) -
Salzman Michele Marie,
Warner Edgewood R.,
Riess Matthias L.
Publication year - 2016
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.30.1_supplement.718.8
Subject(s) - hypoxia (environmental) , andrology , cell growth , reactive oxygen species , cell , cytotoxicity , cell culture , endothelium , artery , chemistry , oxygen , biology , immunology , medicine , microbiology and biotechnology , in vitro , biochemistry , genetics , organic chemistry
Hypoxic conditions cause extensive cellular damage, particularly to mitochondria and the plasma membrane. Upon reoxygenation, incomplete reduction of oxygen (O 2 ) can produce reactive oxygen species that can cause additional reperfusion injury to damaged and viable cells. Intralipid (IL) has been demonstrated to be cardio‐protective when delivered as a pre‐ or post‐conditioning agent. We hypothesized that IL delivered during reoxygenation will protect human coronary artery endothelial cells (HCAECs) and human cardiomyocytes (HCMs) from reoxygenation (reperfusion) injury. OBJECTIVES We compared 1) hypoxic cells vs hypoxic/reoxygenated (H/R) cells vs hypoxic cells reoxygenated with differing concentrations of IL; and 2) cells on the basis of cellular “age” (passage; e.g., P7 vs P15). METHODS Cell cultures were placed into complete media/normoxic conditions (FBS & glucose; 95% O 2 , 5% CO 2 ) or hypoxic media/hypoxic conditions (0% FBS & glucose; 1% O 2 , 94% N 2 , 5% CO 2 ) at 37°C for 3 hrs. Samples were then either immediately assayed for cell proliferation, metabolic activity, and cytotoxicity, or they were incubated ± IL (0.1%, 0.5%, 1%, or 2%) in complete media/normoxic conditions for 2 hrs and then assayed for the same parameters. RESULTS Younger cells had a greater proliferative capacity than older cells in both cell types. However, there were equivalent decreases in proliferative capacity in both hypoxic and H/R exposed cells, and IL had no effect on cell proliferation, regardless of concentration. Metabolic activity was noted to be higher in older cells than younger cells in both cell types. When exposed to H/R, HCAECs metabolic activity decreased, while HCMs had slightly increased metabolic activity. In both cell types, significant increases in metabolic activity occurred notably with IL concentrations of 1–2%. LDH release was greater in older than in younger cells of both cell types, and was more extensive in hypoxic only cells compared to H/R cells. The dose‐dependent impact of IL on cytotoxicity was indeterminable. CONCLUSIONS Cells differ based on their “age” (passage), and also on what cellular function is affected by H/R. IL increased metabolic activity in both cell types, specifically at concentrations of 1–2%; however, there was no determinable effect on cell proliferation or cytotoxicity. Further studies need be conducted to determine which metabolic pathways are being activated, and whether those pathways are related to cell survival during stressful conditions such as H/R. Support or Funding Information Supported by the Department of Veterans Affairs (IK2BX001278), NIH (R01 HL095122), and institutional funds.