Protective effects of exogenous HS to the recovery of hypoxia post-conditioning on aged H9C2 cells and the underling mechanisms / 中国应用生理学杂志
Chinese Journal of Applied Physiology
; (6): 289-293, 2018.
Article
in Zh
| WPRIM
| ID: wpr-773757
Responsible library:
WPRO
ABSTRACT
OBJECTIVE@#To investigate the recovery of protective effects of exogenous hydrogen sulfide (HS) on hypoxia post-conditioning in aged H9C2 cells and its mechanism.@*METHODS@#H9C2 cells (cardiomyocytes line) were treated with 30 μmol/L hydrogen peroxide (HO) for 2 hours, then cultured for 3 days in order to induce cellular aging. Aged H9C2 cells were randomly divided into 5 groups (=8)Control group (Control), hypoxia/reoxygenation group (H/R), H/R + NaHS group, hypoxia post-conditioning (PC) group, PC+NaHS group. H/R modelthe cells were exposed to hypoxic culture medium (serum and sugar free medium, pH=6.8) for 3 hours and then cultured at normal condition for 6 hours. PC modelat the end of hypoxia for 3 hours, the cells were exposed to normoxic culture solution for 5 minutes, then the cells were placed in hypoxic solution for 5 minutes, the cycle above-mentioned was repeated 3 times and followed by reoxygenation for 6 hours. Advanced glycation end products (AGEs) content and caspase-3 activity were detected by ELISA. The cell viability was observed by cell counting kit-8 (CCK-8). The reactive oxygen species (ROS) levels were analyzed using 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. The apoptotic rate was determined through Hoechst 33342 staining. The mRNA levels of relative gene expression were detected by real-time PCR.@*RESULTS@#Thirty μmol/L HO induced H9C2 cell senescence while did not lead to apoptosis. Compared with control group, cell viability was decreased, the apoptotic rate、levels of ROS and the mRNA of caspase-3, caspase-9 and Bcl-2 were increased in H/R and PC groups (<0.01). There were no differences in the above indexes between PC group and H/R group. Supplementation of NaHS increased cell viability and decreased apoptotic rate and oxidative stress. The effects of PC + NaHS on the above indexes were better than those of H/R+NaHS group.@*CONCLUSIONS@#Exogenous HS can restore the protective effect of PC on the aged H9C2 cells, and its mechanism is related to the inhibition of oxidative stress and apoptosis.
Key words
Full text:
1
Index:
WPRIM
Main subject:
Cell Hypoxia
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Cell Survival
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Reactive Oxygen Species
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Apoptosis
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Myocytes, Cardiac
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Hydrogen Peroxide
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Applied Physiology
Year:
2018
Type:
Article