Ionomycin calcium salt

Cell lines

N1E-115 cells

Preparation Method

Cells were treated with various concentrations (0, 0.2, 0.5, 1, 2, and 10 µM) and incubation times (3, 6, 12, and 24 h) of ionomycin, and cell survival was determined using the trypan blue dye exclusion assay.

Reaction Conditions

0, 0.2, 0.5, 1, 2, and 10 µM；3, 6, 12, and 24 h

Applications

Ionomycin treatment induced cell death in a concentration- and time-dependent manner.

Animal models

Adult male C57BL/6 wild type (20-25 g, 8-10 weeks old) and FXR KO mice (20-25 g, 8-10 weeks old)

Preparation Method

In order to investigate the biological function of FXR in vivo, an L-type calcium channel agonist, Bayk8644 (2 mg/kg) was given intraperitoneally at 10 min after reperfusion. Bayk8644 stock solution (1 mmol/L in dimethyl sulfoxide, DMSO) was diluted in 1× phosphate-buffered saline (PBS) with a final DMSO concentration of 4.6%. Similarly, PBS with 4.6% DMSO was given intraperitoneally in sham, wild-type vehicle, and FXR knockout vehicle groups as control. A dose range of ionomycin (50 nmol/L), GW4064 (0, 1, 1.5, 3, and 6 µmol/L), or Z-guggulsterone (Z-GS, 0, 2.5, 5, and 10 µmol/L) was given for 18 h before oxygen and glucose deprivation (OGD).

Dosage form

50 nmol/L (4.6% DMSO); 18 h; i.p.

Applications

TUNEL staining demonstrated that Z-GS treatment reduced neuronal apoptosis and addition of IM (Ionomycin) significantly increased apoptotic neurons compared to OGD group, suggesting that inhibition of FXR (Farnesoid X receptor) can reduce neuronal apoptosis through reducing calcium influx after OGD/R.

References:

Nakamura S, et al. Ionomycin-induced calcium influx induces neurite degeneration in mouse neuroblastoma cells: analysis of a time-lapse live cell imaging system. Free Radic Res. 2016;50(11):1214-1225.
Shan HM, et al. Farnesoid X receptor knockout protects brain against ischemic injury through reducing neuronal apoptosis in mice. J Neuroinflammation. 2020 May 25;17(1):164.