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Xinshuaining preparation protects H9c2 cells from H2O2-induced oxidative damage through the PI3K/Akt/Nrf-2 signaling
Mingjun Han1, Jie Lin1, Yi Yang1
1Department of Cardiology, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu Sichuan, PR China.
Insights
Xinshuaining preparation protects H9c2 cells from oxidative stress by increasing cell viability and reducing apoptosis. This effect is mediated through the PI3K/Akt/Nrf-2 signaling pathway, highlighting its therapeutic potential for cardiovascular diseases.
Area of Science:
- Cardiovascular Pharmacology
- Cellular Biology
- Oxidative Stress Research
Background:
- Cardiovascular disease (CVD) is a leading cause of mortality globally.
- Oxidative stress is a key pathological mechanism implicated in various CVDs.
- Xinshuaining preparation demonstrates therapeutic effects in heart failure, but its antioxidant role in H9c2 cells is not well-defined.
Purpose of the Study:
- To investigate the anti-oxidative stress effects of Xinshuaining preparation on H9c2 cells.
- To elucidate the underlying molecular mechanisms, particularly the involvement of the PI3K/Akt/Nrf-2 signaling pathway.
- To assess the impact of Xinshuaining on cell viability, apoptosis, and oxidative stress markers in a cellular model.
Main Methods:
- H9c2 cells were exposed to hydrogen peroxide (H2O2) to induce oxidative stress.
- Medicated serum of Xinshuaining preparation was applied, and its effects were analyzed.
- Key indicators evaluated included cell viability (CCK-8), apoptosis rate, oxidative stress markers (SOD, GSH-Px, MDA), mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) levels.
- Protein expression levels of PI3K, Akt, and Nrf-2 were assessed via Western blot, with LY294002 used to inhibit the PI3K/Akt pathway.
Main Results:
- Xinshuaining preparation significantly enhanced H2O2-impaired cell viability and mitochondrial membrane potential.
- It increased the levels of antioxidant enzymes (SOD, GSH-Px) and the expression of phosphorylated PI3K, Akt, and Nrf-2.
- Conversely, Xinshuaining reduced H2O2-induced apoptosis, MDA levels, and ROS production.
- The protective effects were partially blocked by LY294002, indicating PI3K/Akt pathway involvement.
Conclusions:
- Xinshuaining preparation exhibits significant protective effects against oxidative stress in H9c2 cells.
- The mechanism involves the activation of the PI3K/Akt/Nrf-2 signaling pathway.
- These findings support the potential of Xinshuaining preparation as a therapeutic agent for cardiovascular conditions associated with oxidative stress.
Background:
Cardiovascular disease (CVD) is the leading cause of death. Oxidative stress is an important pathological process of a variety of CVDs. Xinshuaining preparation has a therapeutic effect on the heart failure. However, the anti-oxidative stress role of Xinshuaining preparation in H9c2 cells is still unclear.
Methods:
The medicated serum of Xinshuaining preparation was acquired and utilized to hatch with H2O2-induced H9c2 cells. Main components in the Xinshuaining preparation were analyzed by liquid chromatography-mass spectrometry (LC/MS). The effect of medicated serum on the cell viability, apoptosis rate, the oxidative stress indicators (SOD, GSH-Px, and MDA), mitochondrial membrane potential (MMP), and ROS level was evaluated by CCK-8, flow cytometry, commercial biochemical detection kits, and JC-1 staining. Additionally, the associated mechanism was determined by the detection of the protein levels (PI3K, phosphorylated PI3K, Akt, phosphorylated Akt, and Nrf-2) through western blot assays, which was also further assessed with the application of LY294002.
Results:
The medicated serum of Xinshuaining preparation notably increased the H2O2-reduced, the cell viability, the concentration of SOD and GSH-Px, MMP level and the relative protein expression level of phosphorylated PI3K and Akt and Nrf-2, while dampened the H2O2-elevated the level of the cell apoptosis rate, MDA, and ROS. However, Xinshuaining preparation on the cell viability, apoptosis, and oxidative stress was notably antagonized by LY294002 pre-treatment.
Conclusions:
The medicated serum of Xinshuaining preparation increased the cell viability and suppressed apoptosis and oxidative stress via the PI3K/Akt/Nrf-2 signaling pathway.
