Glp 1

For CML (MyBioSource, San Diego, USA, Catalogue No.: MBS390113) stimulating, cells (5×103) in 100 μl DMEM were added to 96-well plates with different concentrations of CML (0, 12.5, 25, 50, 75, and 100μg/ml) for 6, 12 and 24 hours. GLP-1 (Bioss, Beijing, China, Catalogue No.: bs-0038P) treatment was performed with GLP-1 at concentrations of 0nM, 50nM, 100nM, 200nM, and 500nM in the medium with cells stimulated by 50 μg/ml CML for 24 hours. GW9662 (MedChemExpress LLC, Shanghai, China, Catalogue No.: 22978-25-2) blocking administration was conducted by 0, 1, 5, 10, 20, and 50 μM GW9662 in PC12 cells with 50 μg/ml CML and 100nM GLP-1 for 24 hours. After CML stimulating, GLP-1 treatment and GW9662 blocking assay, CCK-8 assay was performed according to the protocol of manufacturer (Jiangsu KeyGEN BioTECH Corp., Ltd, Nanjing, China, Catalogue No.: KGA317-2).

DON (purity >98%) was obtained from Sigma-Aldrich (St. Louis, MA, USA). DON was dissolved in sterile phosphate buffered saline (PBS) containing 1% dimethyl sulfoxide (DMSO) and dissolved to 0.1, 0.5, 1, 2.5, and 5 mg/kg·bw. GIP (MedChemExpress; Trenton, NJ, USA), PYY, and CCK (Sigma-Aldrich; St. Louis, MO, USA) were respectively diluted in PBS, at a dose of 0.1 mg/kg·bw. GLP-1 (MedChemExpress) was made into a 0.01 mg/kg·bw solution with PBS. The PYY receptor antagonist JNJ-31020028, the CCK receptor antagonist Proglumide (Sigma-Aldrich), and GLP-1 receptor antagonist Exendin(9-39) were respectively dissolved in 100 μL sterile PBS, so that their final doses were 10, 200, or 0.1 mg/kg·bw. GIP receptor inhibition GIP(3-30)NH₂ was synthesized by the Syn peptide (Shanghai, China), diluted with normal saline to 50 nm/kg·bw, and injected through the tail vein. According to the instructions of the reagent supplier, previous animal research and thesis data, the dosages of various pharmacological agents were selected [27 (link),31 (link),34 (link),35 (link)].