堇叶碎米荠富硒多肽对乙醇诱导THLE-2肝细胞损伤的保护作用

Protective effect of selenium-enriched peptide from Cardamine violifolia on ethanol-induced THLE-2 hepatocyte injury

采用堇叶碎米荠富硒多肽(selenium-enriched peptide of Cardamine violifolia,SPE)处理人源肝细胞,研究SPE能否干预由乙醇引起的肝细胞损伤。利用酒精诱导损伤THLE-2肝细胞构建氧化应激模型,通过不同浓度有机硒的SPE处理后,测定细胞损伤和氧化应激相关指标,评价其细胞保护和抗氧化能力。结果显示,乙醇浓度为1000 mmol/L,乙醇作用时间为12 h时,可对THLE-2细胞造成明显损伤(P<0.05)。SPE处理可明显提高细胞存活率(P<0.05),在12μmol/L硒浓度作用下可达78.02%。与损伤组相比较,SPE处理组上清液的谷草转氨酶、谷丙转氨酶和乳酸脱氢酶含量显著减少(P<0.05),并且,SPE处理后,THLE-2细胞内活性氧含量减少,超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶和谷胱甘肽水平显著上升(P<0.05),最高分别为34.68 U/mg、974.75 nmol/g、1.76 U/mg和55.58μmol/g,其中,谷胱甘肽过氧化物酶高度表达,约为损伤组的5.6倍,可能因为SPE为谷胱甘肽过氧化物酶合成提供硒离子。另外,SPE处理组的细胞凋亡率降低,线粒体膜电位升高。综上所述,堇叶碎米荠富硒多肽可以通过调控氧化应激相关指标来达到缓解酒精性肝损伤的目的。

Human hepatocytes were treated with a selenium-enriched peptide from Cardamine violifolia(SPE)to study whether SPE could interfere with ethanol-induced hepatocyte injury.An oxidative stress model was constructed via the alcohol-induced injury of THLE-2 hepatocytes.After treatment with SPE at different concentrations of organic selenium,the cell injury and oxidative stress related indicators were measured to evaluate the cell protection and antioxidant capacity.The results showed that the concentration of ethanol of 1000 mmol/L and the action time of ethanol of 12 h led to significant damage of THLE-2 cells(P<0.05).SPE treatment significantly increased the cell viability(P<0.05),which reached 78.02%at selenium concentration of 12μmol/L.Compared with the injury group,the contents of aspartate aminotransferase,alanine aminotransferase,and lactate dehydrogenase in the supernatant of SPE treatment group was significantly reduced(P<0.05).Moreover,after SPE treatment,the content of reactive oxygen species in THLE-2 cells was decreased,and the levels of superoxide dismutase,glutathione peroxidase,catalase,and glutathione were significantly increased(P<0.05),with the highest values of 34.68 U/mg,974.75 nmol/g,1.76 U/mg,and 55.58μmol/g,respectively;among them,glutathione peroxidase was highly expressed,at about 5.6 times that of the injury group,probably because SPE provided selenium ions for glutathione peroxidase synthesis.In addition,the apoptosis rate was decreased,and the mitochondrial membrane potential was increased in the SPE treatment group.Above all,the selenium-enriched polypeptide of Cardamine violifolia alleviated alcoholic liver injury by regulating oxidative stress related indicators.