RESUMO
ObjectiveTo investigate the effects of flavanomarein on the transcriptome of small intestinal organoids in insulin-resistant mice. MethodFirstly， small intestinal organoids of C57BL/6J and db/db mice were established. Ki-67 and E-cadherin expression was determined by immunofluorescence. Small intestinal organoids were divided into the following three groups： C57BL/6J mouse small intestinal organoids as the normal control group， db/db mouse small intestinal organoids as the model group （IR group）， and db/db mouse small intestinal organoids treated with flavanomarein as the administration group （FM group）. Western blot was used to detect the expression of glucagon-like peptide-1（GLP-1） protein on the small intestinal organoids of the three groups. Finally， transcriptome sequencing was performed on samples from the three groups. ResultOn the 6th day of small intestine organoids culture， a cyclic structure was formed around the lumen， and a small intestine organoids culture model was preliminarily established. Immunofluorescence detection showed that ki-67 and E-cadherin were expressed in small intestinal organoids. Western blot results showed that the expression of GLP-1 protein was increased by flavanomarein. In the results of differential expressed gene （DEG） screening， there were 1 862 DEGs in the IR group as compared with the normal control group， and 2 282 DEGs in the FM group as compared with the IR group. Through protein-protein interaction（PPI） network analysis of the DEGs of the two groups， 10 Hub genes， including Nr1i3， Cyp2c44， Ugt2b1， Gsta1， Gstm2， Ptgs1， Gstm4， Cyp2c38， Cyp4a32， and Gpx3， were obtained. These genes were highly expressed in the normal control group， and their expression was reduced in the IR group. After the intervention of flavanomarein， the expression of the above genes was reversed. ConclusionFlavanomarein may play its role in improving insulin resistance by reversing the expression levels of 10 Hub genes， including Nr1i3， Cyp2c44， Ugt2b1， Gsta1， Gstm2， Ptgs1， Gstm4， Cyp2c38， Cyp4a32， and Gpx3.
RESUMO
Objective To establish a small intestinal organoid culture system as an in vitro study model of intesti-nal epithelial cells, and to explore the relevant pathological detection techniques and provide a convenient platform for in vitro study of various intestinal diseases. Methods The mouse intestinal epithelium was isolated and cultured into or-ganoids to simulate the growth and development of intestinal epithelium in vitro. The proliferation and differentiation signals were detected by immunohistochemistry and three-dimensional immunofluorescence technique. Results The culture system of the mouse small intestine epithelium was established. Immunohistochemical staining and three-dimensional immunofluo-rescence technique were successfully used to detect the growth and development of small intestinal organoids. Conclusions The successfully established mouse small intestinal organoid culture system and application of immunoassay technology will gradually become a most favorable technical means for studies of various intestinal diseases.
RESUMO
Objective To establish a small intestinal organoid culture system as an in vitro study model of intesti-nal epithelial cells, and to explore the relevant pathological detection techniques and provide a convenient platform for in vitro study of various intestinal diseases. Methods The mouse intestinal epithelium was isolated and cultured into or-ganoids to simulate the growth and development of intestinal epithelium in vitro. The proliferation and differentiation signals were detected by immunohistochemistry and three-dimensional immunofluorescence technique. Results The culture system of the mouse small intestine epithelium was established. Immunohistochemical staining and three-dimensional immunofluo-rescence technique were successfully used to detect the growth and development of small intestinal organoids. Conclusions The successfully established mouse small intestinal organoid culture system and application of immunoassay technology will gradually become a most favorable technical means for studies of various intestinal diseases.