The mechanism of hyperoside protection of ECV-304 cells against tert-butyl hydroperoxide-induced injury.
The aim of the present study was to investigate the mechanism of hyperoside protecting ECV-304 cells against tertbutyl hydroperoxide (TBHP)-induced injury. ECV-304 cell viability was measured by MTT assay. Cellular morphologic changes were observed using phase contrast microscopy. The genotoxic effects of TBHP and the protective ability of hyperoside were assessed by the Comet test. Lipid peroxidation was measured by HPLC method. The cellular redox status was determined from GSH/GSSG ratios. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. Western blot analysis was used to evaluate the levels of cytochrome c, p53, SIRT1, Bax and Bcl-2 expression. The results showed that 128 mumol/l hyperoside could effectively protect TBHP-treated ECV-304 cells from death, increase superoxide dismutase activity and significantly decrease malondialdehyde production. Hyperoside was effective in protecting against the induction of oxidized DNA bases and redox state alterations induced by TBHP. Furthermore, the release of proapoptotic cytochrome c from mitochondria was reduced by hyperoside, which increased the expression of antiapoptotic SIRT1 and inhibited the translocation of Bax from cytoplasm to mitochondria. Taken together, these results indicate that hyperoside is effective in protecting against the oxidative damage induced by TBHP. The mechanism of hyperoside protecting against ECV-304 cell apoptosis by TBHP is related with resuming mitochondrial function and regulating the expression of SIRT1 and Bcl-2 family members.
School of Basic Medical Sciences, Liaoning University of TCM, Shenyang, China.
Hai Bo
HB
Li
Xin
X
Yi
Jian Mei
JM
Gao
Xi Xiang
XX
Ying
Hong Quan
HQ
Guan
Jian Chun
JC
Li
D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects*; D008858:Microscopy, Phase-Contrast; D008928:Mitochondria / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000235:genetics; D011794:Quercetin / Q000031:analogs & derivatives* / Q000494:pharmacology; D056564:Sirtuin 1; D037761:Sirtuins / Q000187:drug effects / Q000235:genetics; D020122:tert-Butylhydroperoxide / Q000633:toxicity
D019253:Proto-Oncogene Proteins c-bcl-2; C021304:hyperoside; D020122:tert-Butylhydroperoxide; D011794:Quercetin; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D037761:Sirtuins