青岛大学学报（医学版）2023，Vol.59Issue(6)：867-873,7.DOI:10.11712/jms.2096-5532.2023.59.200
岩藻多糖对高尿酸诱导Hep G2细胞凋亡影响及机制
EFFECT OF FUCOIDAN ON HEPG2 CELL APOPTOSIS INDUCED BY HIGH URIC ACID AND ITS MECHANISM
摘要
Abstract
Objective To investigate the effect of fucoidan on mitochondrial apoptosis in human hepatoma(HepG2)cells induced by high concentration of uric acid(UA)and its mechanism.Methods HepG2 cells were randomly divided into control group(added with drug-free culture solution),UA model group(added with 0.2 g/L UA solution),resveratrol group(Res group,pretreated with 1 μmol/L resveratrol solution for 24 h,and then added with 0.2 g/L UA solution),F1 group(added with 0.2 g/L UA + 20 mg/L fucoidan),F2 group(added with 0.2 g/L UA + 40 mg/L fucoidan),F1+EX527 group(added with 0.2 g/L UA + 20 mg/L fucoidan + 1μmol/L EX52),and F2+EX527 group(added with 0.2 g/L UA + 40 mg/L fucoidan + 1μmol/L EX527).After 24 h treatment with corresponding drugs,Cell Counting Kit-8 assay was used to determine the viability of HepG2 cells;enzyme-linked immunosorbent assay was used to measure alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels in the cell supernatant,as well as intracellular superoxide dismutase(SOD),reduced glutathione(GSH),and ma-londialdehyde(MDA)levels;fluorescence microscopy was used to measure reactive oxygen species(ROS)production;flow cy-tometry was used to determine the cell apoptosis rate,and Western blot was used to measure the expression of Sirt1.Results Compared with the control group,the UA model group had significantly lower HepG2 cell viability(F=295.200,P<0.01),SOD and GSH levels(F=47.880,8.261;P<0.01),and Sirt1 protein expression(F=64.520,P<0.01),and significantly higher ALT and AST levels in cell supernatant(F=204.300,9.511;P<0.01),MDA content(F = 132.400,P<0.01),ROS production(F = 23.720,P<0.05),and cell apoptosis rate(F=19.200,P<0.01).Compared with the UA model group,the Res group,F1 group,and F2 group had significantly higher HepG2 cell survival rates and SOD and GSH levels(all P<0.01),and significantly lower ALT and AST levels and MDA content(all P<0.01).Compared with the UA model group,the F2 group had significantly higher Sirt1 expression,and significantly lower intracellular ROS production,mi-tochondrial membrane potential,and cell apoptosis rate(P<0.05).The addition of Sirt1 inhibitor EX527 significantly reversed the antioxidant and anti-apoptotic effects of fucoidan(P<0.05).Conclusion Fucoidan can inhibit mitochondrial oxidative stress and reverse UA-induced mitochondrial apoptosis in HepG2 cells by up-regulating Sirt1 expression.关键词
尿酸/岩藻多糖/癌,肝细胞/氧化性应激/细胞凋亡Key words
uric acid/fucoidan/carcinoma,hepatocellular/oxidative stress/apoptosis分类
医药卫生引用本文复制引用
裴忠仟,薛美兰,杨佳,张楠,高海琪..岩藻多糖对高尿酸诱导Hep G2细胞凋亡影响及机制[J].青岛大学学报（医学版）,2023,59(6):867-873,7.基金项目
国家自然科学基金资助项目(81872605) （81872605）
山东省自然科学基金资助项目(ZR2020MH215) （ZR2020MH215）
