Deoxycholic acid sodium salt (Sodium deoxycholate)|302-95-4|安捷凯

Background

Deoxycholic acid (DCA) is a secondary bile acid that is formed via microbial transformation of cholic acid in the colon.¹ It can be conjugated to glycine or taurine to produce glycodeoxycholic acid or taurodeoxycholic acid , respectively, in hepatocytes.^1,2,3 DCA (0.2% v/v) inhibits spore germination induced by taurocholic acid in seven C. difficile strains, as well as inhibits growth and decreases the cytotoxicity of C. difficile culture supernatants to Vero cells when used at a concentration of 0.02% v/v.¹ It inhibits ionizing radiation-induced p53-dependent transcription in a reporter assay using HCT116 cells when used at a concentration of 200 ?M.⁴ Fecal and intestinal tissue levels of DCA are increased in a rat model of high-fat diet-induced obesity compared with rats fed a normal diet.⁵ Increased serum DCA levels have been found in patients with colorectal cancer.⁶

1.Thanissery, R., Winston, J.A., and Theriot, C.M.Inhibition of spore germination, growth, and toxin activity of clinically relevant C. difficile strains by gut microbiota derived secondary bile acidsAnaerobe4586-100(2017) 2.Schmid, A., Neumann, H., Karrasch, T., et al.Bile acid metabolome after an oral lipid tolerance test by liquid chromatography-tandem mass spectrometry (LC-MS/MS)PLoS One11(2)e0148869(2016) 3.?arenac, T.M., and Mikov, M.Bile acid synthesis: From nature to the chemical modification and synthesis and their applications as drugs and nutrientsFront. Pharmacol.9939(2018) 4.Qiao, D., Gaitonde, S.V., Qi, W., et al.Deoxycholic acid suppresses p53 by stimulating proteasome-mediated p53 protein degradationCarcinogenesis22(6)957-964(2001) 5.Lin, H., An, Y., Tang, H., et al.Alterations of bile acids and gut microbiota in obesity induced by high fat diet in rat modelJ. Agric. Food Chem.67(13)3624-3632(2019) 6.Bayerd?rffer, E., Mannes, G.A., Richter, W.O., et al.Increased serum deoxycholic acid levels in men with colorectal adenomasGastroenterology104(1)145-151(1993)

Protocol

Cell experiment:

MGC803 cells are cultured in Roswell Park Memorial Institute media supplemented with 10% fetal calf serum and 100 U/mL Penicillin and 100 mg/mL Streptomycin. To generate MGC803-resistant cells, the pH value of the MGC803 culture medium is adjusted to the experimental conditions using the hydrochloric acid (A). The bile acids GCDA and Deoxycholic acid are diluted to optimal working concentrations of 100 μM (B) with culture medium, and the overall pH (A+B) is adjusted to pH 5.5, simulating the gastric environment. Initially, MGC803 cells are chronically exposed to acidified medium with bile acids (A+B) for 10 min every 24 h. The experimental time and conditions are optimized in our preliminary experiments, which show that 10 min is enough and does not result in cell damage. This procedure is repeated and it takes 60 weeks for the MGC803 cells to survive and proliferate under the exposure of A+B for 120 min. Control untreated cells are cultured in neutral RPMI medium at pH 7.4 in parallel to the resistant cells for 60 weeks. The morphological changes in MGC803 cells exposed to acidified bile acids (A+B) are documented at 30 and 60 weeks[2].

参考文献:

[1]. Somm E, et al. β-Klotho deficiency protects against obesity through a crosstalk between liver, microbiota, and brown adipose tissue. JCI Insight. 2017 Apr 20;2(8). pii: 91809.
[2]. Wang X, et al. Acidified bile acids enhance tumor progression and telomerase activity of gastric cancer in micedependent on c-Myc expression. Cancer Med. 2017 Apr;6(4):788-797.