Cell lines

SGBS preadipocytes

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while.Stock solution can be stored below -20°C for several months.

Reaction Conditions

10 μg/ml, 9 hours

Applications

Addition of CHX enhanced CD95-induced cleavage of caspase-8 into p43/p41 intermediate and p18 active fragments as well as proteolytic turnover of the proenzyme form of caspase-8 after 1, 3, 6, and 9 h. In addition, CHX increased cleavage of caspase-3 into the active p20/17 fragment at these time points. At later time points (24, 48, and 72 h), a decrease in the p55 pro-form of caspase-8 and the p35 pro-form of caspase-3 was observed. Interestingly, α-APO-1 alone induced caspase-8 and caspase-3 cleavage (3, 6, 9 h) although there is no induction of cell death after 24 h. Involvement of caspase-cleavage was confirmed by the use of the caspase inhibitor Z-VAD.fmk, which reduced CD95- and CHX-induced apoptosis. Apoptosis was rescued by ~50% pointing to a potential role of caspase-independent cell death in SGBS preadipocytes.

Animal models

Sprague Dawley rat pups

Dosage form

Intraperitoneal injection, 0.6 mg/kg, 0, 6, 12 or 24 hr

Applications

The hypoxia-ischemia model was set up using the rat pups. The hypoxia-ischemia control group (HI) and hypoxia-ischemia were treated with cycloheximide treatment group at 0, 6, 12, 24 and 24 hr after HI (HI_0, 6, 12, 24), respectively. Infarct volume, as measured by morphometric analysis of infarct areas with TTC, was significantly reduced by 92% and 61% when cycloheximide was given 0 or 6 hr after HI respectively, but showed an insignificant trend in infarct reduction if cycloheximide was administered 12 hr after HI compared to the HI control group, and no protective effect was observed when administration was delayed until 24 hr after HI.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1] Fischer-Posovszky P, Keuper M, Nagel S, et al. Downregulation of FLIP by cycloheximide sensitizes human fat cells to CD95-induced apoptosis. Experimental cell research, 2011, 317(15): 2200-2209.

[2] Park W S, Sung D K, Kang S, et al. Therapeutic window for cycloheximide treatment after hypoxic-ischemic brain injury in neonatal rats. Journal of Korean medical science, 2006, 21(3): 490-494.