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Cystatin C alleviates H2O2-induced H9c2 cell injury
1Department of Cardiology, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China. suqiang1983@foxmail.com.
Insights
Cystatin C (Cys C) effectively treats myocardial ischemia-reperfusion injury (MIRI) by reducing cardiomyocyte oxidative stress and apoptosis. This study demonstrates Cys C
Area of Science:
- Cardiovascular Research
- Cellular Biology
- Biochemistry
Background:
- Acute myocardial infarction incidence is rising, posing a significant mortality risk.
- Myocardial ischemia-reperfusion injury (MIRI) complicates treatment with no effective clinical solutions.
- Investigating novel therapeutic targets for MIRI is crucial.
Purpose of the Study:
- To investigate the therapeutic effects of Cystatin C (Cys C) on myocardial ischemia-reperfusion injury (MIRI).
- To elucidate the mechanisms underlying Cys C's protective effects on cardiomyocytes.
- To evaluate Cys C's efficacy in both in vitro and in vivo MIRI models.
Main Methods:
- H9c2 rat cardiomyocytes were subjected to hydrogen peroxide (H2O2)-induced injury and treated with Cys C.
- Cell viability, oxidative stress markers (LDH, SOD, MDA), and apoptosis-related molecules were assessed.
- The NF-κB signaling pathway activity was analyzed in treated and untreated cells.
- A rat MIRI model was established to validate Cys C's therapeutic potential in vivo.
Main Results:
- Cys C treatment alleviated H2O2-induced H9c2 cell injury, reducing LDH and MDA levels while increasing SOD activity.
- Cys C significantly decreased apoptosis in injured cardiomyocytes.
- Cys C inhibited the elevated NF-κB signaling pathway activity observed in H9c2 cells.
- In vivo studies confirmed Cys C's therapeutic efficacy in a rat MIRI model.
Conclusions:
- Cystatin C mitigates cardiomyocyte oxidative stress and apoptosis.
- The protective mechanism involves the inhibition of the NF-κB signaling pathway.
- Cys C demonstrates significant therapeutic potential for treating MIRI.
Objective:
At present, the incidence of acute myocardial infarction is increasing year by year, and it has become one of the diseases with the highest mortality rate in humans. Myocardial ischemia-reperfusion injury (MIRI) is a major problem in the treatment of myocardial infarction, but clinically there is no effective way to treat MIRI. This study used Cystatin C (Cys C) to treat cardiomyocytes and rats to investigate the effect of Cys C on MIRI.
Materials And Methods:
We used H2O2 to induce rat cardiomyocytes (H9c2 cells) injury and stimulated the cells with Cys C. Cell counting kit 8 (CCK8) assay was used to determine the optimal concentration of H2O2 and Cys C to stimulate H9c2 cells. We determined the effects of Cys C on oxidative stress and apoptosis levels in H9c2 cells by measuring the activity of dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA), and the expression of apoptosis-related molecules (caspase3/8/9, Bax and Bcl-2). Changes in the activity of the NF-κB signaling pathway in H9c2 cells were also detected. In addition, we made rat MIRI models by ligating the coronary arteries and used Cys C to treat rats to verify the effect of Cys C on MIRI.
Results:
According to the results of the CCK8 assay, 1000 μM of H2O2 and 15 μM of Cys C were used to stimulate H9c2 cells. Cys C alleviated H2O2-induced H9c2 cell injury, manifested as a decrease in LDH and MDA activity and an increase in SOD activity. Cys C also reduced the apoptosis level in H9c2 cells. The activity of NF-κB signaling pathway in injured H9c2 cells was increased, and stimulation of Cys C could inhibit the NF-κB signaling pathway in H9c2 cells. The application of Cys C in MIRI rats also verified its therapeutic effect on MIRI.
Conclusions:
Cys C reduced the oxidative stress and apoptosis levels of cardiomyocytes by inhibiting the activity of NF-κB signaling pathway in cardiomyocytes, thereby reducing cardiomyocyte injury and treating MIRI.
