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Preventive Effect of Monoammonium Glycyrrhizinate on the Injury Induced by Sodium Oleate in Hepatocytes of Cows in vitro
Authors:ZHANG Cai  SHAO Qi  XU Wenhao  JIA Zhe  MA Lu  TIAN Chao  YANG Zijun
Institution:1. College of Animal Science&Technology, Henan University of Science and Technology, Luoyang 471023, China;2. Henan International Joint Laboratory of Animal Welfare and Health Breeding, Luoyang 471023, China;3. West Anhui University, Liuan 237000, China;4. Henan Vocational College of Agriculture, Zhengzhou 451450, China
Abstract:Hepatocytes injury is the main pathological feature of fatty liver disease in cows during the perinatal period. The aim of this study was to investigate the preventive effect of monoammonium glycyrrhizinate (MAG) on cell damage induced by sodium oleate in hepatocytes of cows. Primary hepatocytes of cows cultured in vitro were divided into 3 groups according to different treatments:control group (untreated primary hepatocytes) and 2 experimental groups (model group, MAG group:add 0 mg·L-1, 0.25 mg·L-1 MAG in culture media respectively, for 12 h and then use sodium oleate at a concentration of 0.25 mmol·L-1 to induce hepatocytes steatosis), and then the activity of hepatocytes were detected by CCK-8, the cell apoptosis rate was calculated by DAPI fluorescence staining, the content of ALT and AST in the culture solution were detected by ultraviolet colorimetry, the levels of intracellular fat deposition were analyzed by oil red O staining and imageJ. Real-time fluorescence quantification (RT-qPCR) was used to detect the mRNA expression of lipid metabolism-related genes PPARα,SREBP-1c,ChREBP,CPT1,CPT2,MTP and inflammation-related genes TNF-α,NF-κB,IL-1β,IL-6,IL-8. The experimental results showed that hepatocytes preconditioning with 0.25 mg·L-1 MAG could improve the activity of model cells (P<0.05), and inhibit the apoptosis rate of model cells effectively (P<0.05). The release levels of ALT and AST in the cell culture supernatant of the MAG group were lower than those of the model group (P<0.05). At the same time, compared with the model group, the number of intracellular lipid droplets and the average lipid droplet area of hepatocytes treated with MAG were significantly reduced (P<0.05). And then the levels of lipid metabolism genes ChREBP, PPARα, MTP and inflammation related genes TNF-α, IL-1β, NF-κB, IL-6, IL-8 mRNA were significantly down-regulated in MAG group than the model group (P<0.05). The results showed that MAG could alleviate the impact and relieve lipid deposition induced by sodium oleate in hepatocytes by inhibiting the expression of genes related to lipid metabolism and inflammatory cytokines, which indicated that it could provide protection for the liver.
Keywords:monoammonium glycyrrhizinate  hepatocytes  sodium oleate  inflammation  lipid metabolism  
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