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Published on: October 14, 2016
Etomidate induces cytotoxic effects and gene expression in a murine leukemia macrophage cell line (RAW264.7)
Rick Sai-Chuen Wu1, King-Chuen Wu, Jai-Sing Yang
1Department of Anesthesiology, China Medical University Hospital, and Department of Pharmacology, School of Pharmacy, China Medical University, No 91, Hsueh-Shih Road, Taichung 404, Taiwan, ROC.
Abstract:
Etomidate is an important tool in the arsenal of the emergency physician, and it has been used in a variety of scenarios for both intubation and procedural sedation. In the present study, we investigated the cytotoxicity of etomidate including induction of apoptosis, and levels of protein and gene expressions associated with apoptotic cell death in murine leukemia RAW264.7 cells in vitro. Cytotoxic and apoptotic responses to etomidate of RAW264.7 cells, including cell morphological changes and cell viability were examined and measured by phase-contrast microscopy and flow cytometric assay, respectively. Results indicated that etomidate increased apoptotic cell morphological changes and reduced cell viability in RAW264.7 cells. 4',6-Diamidino-2-phenylindole (DAPI) staining also showed that etomidate induced the formation of apoptotic bodies, a characteristic of apoptosis. Results from Western blotting indicated that etomidate enhanced the levels of cytochrome c, apoptosis-inducing factor (AIF), endonuclease G (Endo G), caspase-9, caspase-3 active form and Bax proteins, but it inhibited the expression of Bcl-xl, leading to apoptosis. DNA microarray assay indicated that etomidate increased the expression of 17 genes (LOC676175; Gm14636; 2810021G02Rik; Iltifb; Olfr1167; Ttc30b; Olfr766; Gas5; Rgs1; LOC280487; V1rd4; Hist1h2bc; V1rj3; Gm10366; Olfr192; Gm10002 and Cspp1) and reduced the expression of 15 genes: (Gm10152; Gm5334; Olfr216; Lcn9; Gm10683; Gm5100; Tdgf1; Cypt2; Gm5595; 1700018F24Rik; Gm10417; Maml2; Olfr591; Trdn and Apol7c). In conclusion, etomidate induced cytotoxic and apoptotic effects the in murine leukemia RAW264.7 cells in vitro.
Insights
Etomidate, a common anesthetic, was found to induce cell death and apoptosis in murine leukemia cells. This study investigated etomidate
Area of Science:
- Cell Biology
- Pharmacology
- Toxicology
Background:
- Etomidate is widely used by emergency physicians for intubation and procedural sedation.
- Understanding the cellular mechanisms of etomidate is crucial for its safe application.
Purpose of the Study:
- To investigate the in vitro cytotoxicity of etomidate on murine leukemia RAW264.7 cells.
- To examine etomidate's role in inducing apoptosis and altering protein and gene expression related to cell death.
Main Methods:
- RAW264.7 cells were treated with etomidate.
- Cytotoxicity and apoptosis were assessed using phase-contrast microscopy and flow cytometry.
- Protein expression was analyzed by Western blotting, and gene expression by DNA microarray.
Main Results:
- Etomidate significantly increased apoptotic morphological changes and decreased cell viability in RAW264.7 cells.
- Western blotting revealed increased levels of pro-apoptotic proteins (cytochrome c, AIF, Endo G, caspase-9, caspase-3, Bax) and decreased Bcl-xl.
- DNA microarray identified 17 upregulated and 15 downregulated genes following etomidate treatment.
Conclusions:
- Etomidate induces significant cytotoxic and apoptotic effects in murine leukemia RAW264.7 cells in vitro.
- The drug modulates key proteins and genes involved in the apoptotic pathway.
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