Application of formononetin to treatment of nonalcoholic fatty liver disease
A formononetin, non-alcoholic technology, applied in the application field of formononetin in the treatment of non-alcoholic fatty liver, can solve the problem of not having formononetin and the like, and achieve the improvement of lipid metabolism disorder , improve insulin resistance
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Embodiment 1
[0057] Embodiment 1: In vivo animal model experiments
[0058] 1. Preparation of high-fat diet (HFD) diet-induced non-alcoholic fatty liver model in mice
[0059] 1.1 Test grouping
[0060] The present invention adopts the method of giving high-fat diet (HFD) to C57BL / 6 mice for 16 weeks to establish a non-alcoholic fatty liver animal model, and verifies its curative effect by intragastric administration of FMN;
[0061] Specific process: The mice were fed with a high-fat diet (HFD), and the fat in the HFD provided 60% of the total calories of the diet. HFD-induced fatty liver is recognized as one of the classic models of NAFLD. Rodents fed HFD for a long time (more than 2 months) will naturally develop hepatic steatosis, and simultaneously develop obesity, hyperlipidemia, and insulin resistance. Inflammation and fibrosis occur less frequently. Select 24 C57BL / 6J male mice, after 7 days of adaptive feeding, they were randomized into normal feeding (ND) group, normal feeding+f...
Embodiment 2
[0087] Embodiment 2: In vitro cell model experiments
[0088] 1) Select the HepG2 liver cancer cell line and primary isolated mouse hepatocytes, stimulate HepG2 cells and primary mouse hepatocytes with 1 mM FFAs for 24 hours, and successfully establish the cell steatosis model, then inject formononetin (20μM ) for 24 h, and at the same time set the BSA group (HepG2 cells were cultured in DMEM low-glucose medium containing 1 % BSA without fetal bovine serum). After the drug intervention, the cells and cell proteins were collected, and the changes in the contents of TG and lipid droplets in the cells were detected later.
[0089] 2) Lipid droplet staining: (1) Cell synchronization: Inoculate HepG2 cells and primary mouse hepatocytes in a 96-well plate (2×103 / well); culture at 37°C, 5% CO2 for 24 h, replace with DMEM Low-glucose complete medium (without fetal bovine serum), cultivated for 12 hours to synchronize the cells in the G0 phase. (2) Dosing: Include HepG2 cells and pri...
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