Effect of genipin on hypoxia/reoxygenation induced oxidative stress injury in H9c2 cell line
- Author:
You-Qin JIANG
1
Author Information
- Publication Type:Journal Article
- Keywords: Genipin; H9c2 cell line; Hypoxia/reoxygenation; Mitochondria; Oxidative stress
- From: Chinese Pharmaceutical Journal 2016;51(1):28-34
- CountryChina
- Language:Chinese
- Abstract: OBJECTIVE: To investigate the effect of genipin on hypoxia/reoxygenation induced oxidative stress injury in H9c2 cell line. METHODS: For mimicking myocardial ischemia/reperfusion (I/R) injury, H9c2 cells were cultured in vitro and established hypoxia/reoxygenation induced oxidative stress injury model. H9c2 cells were divided into six groups: control group, H/R group, H/R+0.5 μmol·L-1 genipin group, H/R+1.25 μmol·L-1genipin group, H/R+2.5 μmol·L-1 genipin group and H/R+10 μmol·L-1 genipin group. Cell viability was detected by cell counting kit-8 assay (CCK-8). The levels of lactate dehydrogenase (LDH), intracellular total superoxide dismutase (T-SOD) and malondialdehyde (MDA) were assessed using microplate reader. Confocal laser scanning microscope was performed to examine the production of reactive oxygen species (ROS) and the level of mitochondrial calcium (m) as well as the depolarization ratio of mitochondrial membranes potential(ΔΨm). The protein expression of cytochrome-C was detected by Western-blot. RESULTS: Comparing to control group, the cell viability of H/R group was significantly decreased in a time-dependent manner(r=-0.82,P<0.01). Low concentration(1.25-40 μmol·L-1) of genipin could improve cell viability exposed to H/R treatment (P<0.05), on the contrary, high concentration(80-320 μmol·L-1) of genipin remarkably reduced the cell viability (P<0.05). In compared with H/R group, the levels of LDH release, MDA production and ROS production, the level of m, depolarization ratio of ΔΨm and the protein expression of cytochrome-c in H/R+(1.25-10 μmol·L-1) genipin groups were notably lessened, while the level of T-SOD was increased(P<0.01 or P<0.05). CONCLUSION: Genipin could reduce H/R-induced oxidative stress injury, the mechanism might be connected with balancing the oxidative stress products and anti-oxidation enzyme system as well as improving mitochondrial dysfunction.
