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10 December 2016 : Laboratory Research  

Chloral Hydrate Treatment Induced Apoptosis of Macrophages via Fas Signaling Pathway

Jun Cai1BE, Yanxia Peng1B, Ting Chen1BC, Huanjin Liao2BE, Lifang Zhang2B, Qiuhua Chen3B, Yiming He3B, Ping Wu2A, Tong Xie3ACD, Qingjun Pan1ACDEG*

DOI: 10.12659/MSM.901772

Med Sci Monit 2016; 22:4836-4843

Abstract

BACKGROUND: There are recent reports on several anesthetics that have anti-inflammatory and anti-infective effects apart from their uses for pain relief and muscle relaxation. Chloral hydrate is a clinical anesthetic drug and sedative that has also been reported to attenuate inflammatory response, but the mechanisms are not clearly understood.

MATERIAL AND METHODS: This study investigated the effect of chloral hydrate treatment on the apoptosis of macrophages and explored the underlying mechanisms. RAW264.7 macrophages were treated with various concentrations of chloral hydrate for various lengths of time. Morphological changes were observed under a light microscope and apoptosis was detected with annexin-V-FITC/PI double-staining assay, Hochest 33258 and DNA ladder assay, the expression of Fas/FasL was detected with a flow cytometer, and the Fas signaling pathway was assessed by Western blotting.

RESULTS: The results showed that chloral hydrate treatment induced the morphology of RAW264.7 macrophages to change shape from typical fusiform to round in a concentration- and time-dependent manner, and was finally suspended in the supernatant. For the induction of apoptosis, chloral hydrate treatment induced the apoptosis of RAW264.7 macrophages from early-to-late stage apoptosis in a concentration- and time-dependent manner. For the mechanism, chloral hydrate treatment induced higher expression of Fas on RAW264.7 macrophages, and was also associated with changes in the expression of proteins involved in Fas signaling pathways.

CONCLUSIONS: Chloral hydrate treatment can induce the apoptosis of RAW264.7 macrophages through the Fas signaling pathway, which may provide new options for adjunctive treatment of acute inflammation.

Keywords: Apoptosis, Chloral Hydrate, Fas Ligand Protein, Macrophages

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750