Method for evaluating the effects of different trace components in wine on cellular autophagy levels

Abstract

The present application is used in the field of food and discloses a method for evaluating the degree of influence of trace components in wine on the level of cellular autophagy, comprising the following steps: Step 1, preparing a single cell suspension of human hepatocytes (LO2) stably expressing GFP-RFP-LC3 protein; Step 2, adding 1.5×10 4 The LO2 cells with a cell / mL were inoculated into a 6-well plate and covered with a glass for 24 hours until they adhered to the wall; Step three, 2 mL of wine containing 80 μL of pure alcohol or wine samples containing different trace components of wine were added to the cells and cultured at 37°C and 5% CO2 for 24 hours. Step four, fix in 4% paraformaldehyde for 15 minutes. Step five, wash the slides with PBS and dry them, then add an anti-fading agent containing DAPI, and place the side covered with cells on the anti-fading agent. Step six, observe the red and green fluorescence using a laser confocal microscope, and judge the effect of the trace components of the wine on the level of cellular autophagy based on the different fluorescence. The present application can quickly evaluate the degree of influence of the trace components in the wine on the level of cellular autophagy by observing the different fluorescence as mentioned above.