Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)
Article Title: Intestinal Gastrin/CCKBR Axis Protects against Type 2 Diabetes by Reducing Intestinal Glucose Absorption through the PI3K/Akt/eIF4B Signaling Pathway.
doi: 10.1002/advs.202410032
Figure Lengend Snippet: Figure 7. Intestinal Gastrin/CCKBR is a negative switch for SGLT1 and GLUT2 expression. A) Principal component analysis (PCA) of protein samples extracted from intestinal epithelial cell (duodenum) villi. (ND-Villin-Cckbr+/+ mice, n = 3; ND-Villin-Cckbr−/−mice, n = 5). B) Pathway mapping analysis using Kyoto Encyclopedia of Genes and Genomes (KEGG); C,D) Western blots and quantification of SGLT1, GLUT2, and GLUT5 protein expressions in the intestinal epithelial cells (duodenum) of ND-Villin-Cckbr+/+ (n = 5); ND-Villin-Cckbr−/−mice (n = 5); HFD-Villin-Cckbr+/+ (n = 5) and HFD-Villin- Cckbr−/−mice (n = 5). E) mRNA levels of Sglt1, Glut2, and Glut5 in intestinal epithelial cells (duodenum) (n = 8 per group). F) Immunofluorescence of SGLT1 (red) using rabbit monoclonal antibody in the duodenum. G) Glucose uptake (absorption) in the isolated small intestines (duodenum) of the mice (n = 5 per group). H) Glucose absorption in isolated duodenum segment (n = 8–9 per group). I,J) SGLT1, GLUT2, and GLUT5 expressions and their quantification in duodenum of ND (fed with normal diet, 10% fat), ND+Gas C57BL/6J mice (ND + 20 mg kg−1 d−1 Gastrin-SiO2 microspheres gavage), HFD (fed with high fat diet, 60% fat), and HFD+Gas C57BL/6J mice (fed with 60% fat diet + 20 mg kg−1 d−1 Gastrin-SiO2 microspheres gavage) (n = 4–6 per group). K) mRNA levels of Sglt1, Glut2, and Glut5 in the small intestines (duodenum) of ND, ND+Gas, HFD, and HFD+Gas C57BL/6J mice; L,M) SGLT1 immunofluorescence staining (DAPI, blue; SGLT1, red) and quantification in the small intestines (duodenum) of ND, ND+Gas, HFD,
Article Snippet: [25] The primary antibody used in the immunochemistry studies was against SGLT1 (203-1-AP, 1:100, Proteintech), whereas the primary antibodies used in the immunofluorescence studies were against villin, CCK, CD31, CD45, EpCAM, gastrin, GIP, GLP-1, PYY, and SGLT1.
Techniques: Expressing, Western Blot, Isolation, Staining