Sarsasapogenin

Cell experiment:

The effect of Sarsasapogenin on cell viability is measured by the MTT assay method. After exposure to Sarsasapogenin in the absence or presence of Z-VAD-FMK pretreatment for the indicated time, cells in 96-well plates are incubated with MTT. The formazan precipitate is dissolved in 200 μL of dimethyl sulfoxide, and the absorbance at 570 nm is measured using a Benchmark microplate reader.

Animal experiment:

The mice are randomly divided into 7 groups: normal control, TNBS-induced colitic control groups treated with vehicle, timosaponin AIII (5 or 10 mg/kg), Sarsasapogenin (5 or 10 mg/kg) or sulfasalazine (50 mg/kg). Each group consists of 6 mice. The colitis is induced by the intrarectal injection of 2.5% (w/v) TNBS solution (100 μL, dissolved in 50% ethanol) into the colon of anesthetized mice using a thin round-tip needle equipped with a syringe. The normal group is treated with vehicle alone instead of TNBS and test agents. If the injected TNBS solution is excreted, the mouse is excluded from the experiment. Timosaponin AIII or Sarsasapogenin (5 or 10 mg/kg) or sulfasalazine (50 mg/kg) dissolved in 2% tween 80 are orally administered once a day for 3 days after treatment with TNBS. The mice are killed 18 h after the final administration of test agents. The colon is quickly removed, opened longitudinally, and gently washed by PBS. Macroscopic evaluation of the colitis grade is scored (0, no ulcer and no inflammation; 1, no ulceration and local hyperemia; 2, ulceration with hyperemia; 3, ulceration and inflammation at one site only; 4, two or more sites of ulceration and inflammation; 5, ulceration extending more than 2 cm). The colons are stored at −80°C until used in the experiment for ELISA and immunoblotting.

References:

[1]. Shen S, et al. Sarsasapogenin induces apoptosis via the reactive oxygen species-mediated mitochondrial pathway and ER stress pathway in HeLa cells. Biochem Biophys Res Commun. 2013 Oct 26.
[2]. Feng B, et al. Sarsasapogenin reverses depressive-like behaviors and nicotinic acetylcholine receptors induced by olfactory bulbectomy. Neurosci Lett. 2017 Feb 3;639:173-178.
[3]. Lim SM, et al. Timosaponin AIII and its metabolite sarsasapogenin ameliorate colitis in mice by inhibiting NF-κB and MAPK activation and restoring Th17/Treg cell balance. Int Immunopharmacol. 2015 Apr;25(2):493-503.