酪蛋白水解物分别与维生素A、维生素C和维生素E协同保护乙醇氧化损伤的肝细胞

Synergistic Protective Effects of Casein Hydrolysate and Vitamins A, C, or E against Ethanol-induced Damage in Hepatocyte HHL-5 Cells

摘要:

酪蛋白水解物可以治疗和修复乙醇氧化损伤的肝细胞。本文的研究目的在于评价维生素A、维生素C和维生素E分别与酪蛋白水解物（Casein Hydrolysate，CH）协同保护乙醇氧化损伤的肝细胞。已有研究表明，酪蛋白水解物对肝细胞HHL-5没有明显的毒性，甚至有着良好的促进增殖的结果。结果表明，300 mmoL/L的乙醇对细胞有着明显的损伤作用；酪蛋白水解物显著提高了乙醇损伤肝细胞HHL-5的细胞存活率。分别在维生素A（0.0344 mg/mL）、维生素C（0.0881 mg/mL）和维生素E（0.1077 mg/mL）的浓度下与酪蛋白水解物协同作用显著提高了乙醇损伤细胞的细胞存活率。通过CCK-8法和培养基内LDH含量的测定，选取最佳CH的协同浓度。通过胞内丙二醛含量的测定和胞内ROS含量的测定进行再次验证。2 mg/mL的酪蛋白水解物单独作用乙醇氧化损伤的细胞有着一定的修复作用，且该浓度下分别与维生素A、维生素C和维生素E协同保护作用显著增强（p<0.05）。

Abstract:

Casein hydrolysate (CH) can be used to treat and repair hepatocytes damaged by ethanol-induced oxidative stress. The aim of this study was to evaluate the synergistic effects of vitamin A, vitamin C, or vitamin E with CH to protect the liver cells from oxidative damage in vitro. Previous studies have shown that CH has no obvious toxicity to liver cells (HHL-5), and can even promote proliferation. The results showed that 300 mmol/L ethanol induced obviously oxidative damage to HHL-5 cells, and CH significantly enhanced the viability of these cells. Combining vitamin A (0.0344 mg/mL), vitamin C (0.0881 mg/mL), or vitamin E (0.1077 mg/mL) with CH significantly enhanced the viability of liver cells damaged by ethanol-induced oxidation. The concentration of CH for the optimum synergistic effect was selected using a cell counting kit-8 (CCK-8) assay and determination of lactate dehydrogenase (LDH) content in the culture medium. This concentration was verified through the determination of intracellular content of malondialdehyde (MDA) and reactive oxygen species (ROS). CH at 2 mg/mL alone exhibited a certain restorative effect on cells damaged by ethanol-induced oxidation, and the synergistic protective effects of combining CH at this concentration with vitamin A, vitamin C, or vitamin E were significantly higher (p <0.05).