摘要
目的探讨乌司他丁(UTI)预处理对HepG2细胞氧化应激损伤的保护作用。方法将HepG2细胞常规培养传代,随机分为H2O2组,UTI100+H2O2组、UTI500+H2O2组、UTI1000+H2O2组及对照组,其中对照组不作任何处理,H2O2组,UTI100+H2O2组、UTI500+H2O2组、UTI1000+H2O2组分别加入浓度为0μmol/L、100μmol/L、500μmol/L、1000μmol/LUTI预处理,24h后加入300μmol/LH2O2处理4h,体外模拟肝细胞损伤。CCK-8试剂盒和LDH试剂盒检测LDH变化评价细胞损伤;TUNEL检测细胞凋亡率;Westernblot检测HepG2细胞中LC3-Ⅱ、Cleavedcaspase-3的表达变化情况。结果与H2O2组相比较,CCK-8和LDH检测结果提示,各TUI预处理组能改善细胞生存环境,减轻H2O2对HepG2细胞的应激损伤,提高细胞活力(P<0.05);TUNEL检测提示预处理可降低氧化应激导致的凋亡率(P<0.05);Westernblot结果提示,与H2O2组比较,UTI可上调LC3-Ⅱ的表达(P<0.05),促进细胞自噬的发生,降低Cleaved caspase-3表达,抑制细胞凋亡(P<0.05)。结论UTI可以通过激活LC3-Ⅱ表达,促进细胞自噬,发挥保护作用,拮抗氧化应激导致的细胞凋亡。
Objective To investigate the protective effect of ulinastatin (UTI) preconditioning on oxidative stress injury in HepG2 cells.Methods HepG2 cells were routinely cultured and randomly divided into H2O2 group, UTI100+ H2O2 group, UTI500+ H2O2 group, UTI1000+ H2O2 group and control group. The control group did not do any treatment. H2O2 group, UTI100+ H2O2 group, UTI500+ H2O2 group,UTI1000+ H2O2 group was pretreated with 0 μmol/L, 100 μmol/L, 500 μmol/L, and 1000 μmol/L UTI. After 24 h, 300 μmol/L H2O2 was added for 4 h to simulate hepatocytes in vitro. damage. CCK-8 kit and LDH kit were used to detect LDH changes to evaluate cell damage;TUNEL was used to detect apoptosis rate;Western blot was used to detect the expression of LC3-II and Cleaved caspase-3 in HepG2 cells. Results Compared with the H2O2 group, the results of CCK-8 and LDH showed that each TUI pretreatment group could improve the cell survival environment, reduce the stress damage of H2O2 on HepG2 cells, and increase cell viability (P<0.05). TUNEL detection suggested pretreatment. The apoptosis rate induced by oxidative stress was reduced (P<0.05).Western blot results showed that compared with H2O2 group, UTI could up-regulate the expression of LC3-II (P<0.05), promote the occurrence of autophagy, decrease the expression of Cleaved caspase-3, and inhibit cell apoptosis (P<0.05). Conclusion UTI can promote the autophagy of cells by activating LC3-II expression, and play a protective role in antagonizing apoptosis induced by oxidative stress.
作者
杨巧侠
王成果
王元欣
YANG Qiao-xia;WANG Cheng-guo;WANG Yuan-xin(Department of Gastroenterology,Fufeng County People's Hospital,Baoji 722200,Shaanxi,China;Department of genealsurgery,Tangdu Hospital, Air Force Military Medical University,Xi’an,710038,Shaanxi,China;Department of Neurosurgery,Fufeng County People's Hospital,Baoji 722200,Shaanxi,China)
出处
《医学信息》
2019年第13期72-76,共5页
Journal of Medical Information
基金
陕西省重点研发计划(编号:2018SF-154)
关键词
乌司他丁
自噬
凋亡
氧化应激
Ulinastatin
Autophagy
Apoptosis
Oxidative stress
