Protective effect of isorhamnetin on H9 C2 cell line against oxidative stress
10.3969/j.issn.1001-1978.2015.06.023
- VernacularTitle:异鼠李素对H2 O2引起的H9 C2细胞氧化应激损伤的保护作用研究
- Author:
Xi DONG
;
Guibo SUN
;
Yun LUO
;
Xiaobo SUN
;
Suhong CHEN
- Publication Type:Journal Article
- Keywords:
isorhamnetin;
myocardial cells;
oxidative stress;
apoptosis;
anti-oxidation;
atherosclerosis
- From:
Chinese Pharmacological Bulletin
2015;(6):853-859,860
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the protective effect of isorhamnetin on H9 C2 myocardial cell line and its mechanisms. Methods The toxicity and optimal pro-tective concentration of isorhamnetin were determined by MTT assay. The experimental subjects were divided into four groups:group N ( normal ) , group M ( mod-el) , group M + ISO ( model + isorhamnetin ) , and group ISO ( isorhamnetin only ) . Group M +ISO and ISO were pre-incubated with isorhamnetin for 12 hours while other groups with plain DMEM. Group M and M+ ISO were treated with 300μmol · L-1 H2 O2 for 4 hours after pre-incubation. Mitochondrial membrane potential depolarization of H9 C2 was measured by fluo-rescence microscope. Apoptotic rate and ROS produc-tion of injured myocardial cell line were detected using
flow cytometry. The oxidative indictors were measured by spectrophotometry. The expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bcl-2, Bax, Nrf2 and HO-1 were examined by Western blot. Result There was no difference in mitochondrial membrane potential depolarization, apoptotic rate, ROS produc-tion, oxidative indictors production and expressions of cytoplasmic cytochrome C, caspase-9,caspase-3, Bcl-2 , Bax between groups ISO and N ( P>0. 05 ) . Apop-totic rate, ROS production, expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bax, MDA pro-duction of group M+ISO were significantly lower than those of group M ( P < 0. 01 ) . And mitochondrial membrane potential, Bcl-2, CAT, SOD and GSH-Px of group M + ISO were increased compared to group M .
Nuclear translocation of Nrf2 and expression of HO-1 in myocardial cell line were increased with the prolonged isorhamnetin incubation time. Conclusion Isorham-netin could protect myocardial cell line against H2 O2-induced oxidative injury and apoptosis through the in-
terruption of mitochondrial dependent apoptotic path-way and activation of Nrf2/ARE pathway.
