Method for detecting anthocyanin-induced rat islet beta cell autophagy effect

Abstract

The invention belongs to the technical field of cytobiology detection and discloses a method for detecting an anthocyanin-induced rat islet beta cell autophagy effect. The method comprises the following steps: preparing an anthocyanin solution, culturing a rat islet beta cell, screening a concentration range of anthocyanin upon the toxicity of the rat islet beta cell, establishing a high-sugar ratislet beta cell apoptosis model by using a flow cytometry technique, performing an active oxygen free radical level experiment and a western blotting method so as to verify the high-sugar rat islet beta cell apoptosis model, detecting the anthocyanin-induced rat islet beta cell autophagy effect by using a cell immumofluorescence method, establishing an RFP-GFP-RIN (red fluorescence protein-greenfluorescent protein-rat islet beta cell) stable cell line to detect the anthocyanin-induced rat islet beta cell autophagy effect, generating autophagosome from the anthocyanin-induced rat islet beta cell, and performing autophagy flux effect judgment. A high-flux screening method is established for the anthocyanin-induced rat islet beta cell autophagy effect, and theoretic bases are provided for preventing diabetes according to the anthocyanin-induced rat islet beta cell autophagy effect.

Description

technical field [0001] The invention belongs to the technical field of cell biology detection, and in particular relates to a method for detecting the autophagy effect of rat islet beta cells induced by anthocyanins. Background technique [0002] At present, the closest existing technology: According to the World Health Organization's forecast, the global diabetes patients will exceed 600 million in 2025, with a growth rate of up to 55%. Globally, diagnosing and treating diabetes costs $82.5 billion annually. Diabetes is mainly divided into type 1, type 2 and gestational diabetes, of which type 2 diabetes accounts for more than 90%. Insulin is secreted in two phases in the body. In people with type 2 diabetes, the insulin in the first phase is higher than normal or normal, and the peak in the second phase is delayed. The decrease in the number of islet β cells and the disorder of their function are closely related to factors such as glucotoxicity and lipotoxicity. Short-t...

AI Technical Summary

Problems solved by technology

[0005] In summary, the existing problems in the prior art are: the ability of different anthocyanin monomers to induce autophagosomes in rat islet β cells in normal and apoptotic models, and the relationship between autophagy ability and structure-activity relationship has not yet been reported.

At present, there is no effective technology to detect the autophagy effect of anthocyanin-induced rat islet β cells, and then evaluate the role of anthocyanin in the prevention of diabetes based on the autophagy effect of anthocyanin-induced rat islet β cells

Images