动物实验,脱氧胆酸 z-bo 2020-08-28 466

　　Objective: To establish a mouse small intestinal organoid culture system and study the effect of deoxycholic acid (DCA) on the growth of small intestinal organoids.

　　Method: Take the terminal ileum of 8 weeks old C57BL/6 mice, separate them with EDTA method, collect the crypts, embed them in Matrigel Matrigel, and culture them in complete medium. A short-term intervention with 100μmol/L high absolute DCA for 2 days and a long-term intervention with 10μmol/LDCA for 10 days were used as a control between absolute alcohol and normal intestinal organs. DCA was taken out and cultured for a long time for 10 days, and the growth and appearance of organoids were observed.

　　Results: Short-term intervention of high-concentration DCA on organoids, the formation rate of globular structure of intestinal organoids, the structure of intestinal organoids [formation rate, evolution rate of organoids and bud number were significantly reduced (P\u003c0.05) ), the short-term clearance index of DCA intravenous infusion still decreased (P \u003cu\→ u0c0.05). After long-term removal of DCA, only intestinal organoid structure formation rate and bud number decreased (P\u003c0.05). Low concentration of DCA interferes with the short-term formation rate of intestinal organoid structure and reduces the number of buds (P\u003c0.05), long-term intervention in intestinal organoids, the formation rate of globular organoids, the structure formation rate of intestinal organoids and The number of buds was significantly reduced (P\u003c0), and after removing DCA, only the structure formation rate and the number of buds of intestinal organoids were lower than usual (P\u003c0). .05).

　　Conclusion: This study successfully established a mouse small intestine organoid culture technique. DCA destroys the small intestine organoids and affects their growth. Long-term deletion of DCA can partially restore its functions.