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. 2022 Sep 21:2022:2483669.
doi: 10.1155/2022/2483669. eCollection 2022.

Protein Kinase N2 Reduces Hydrogen Peroxide-inducedDamage and Apoptosis in PC12 Cells by AntiOxidative Stress and Activation of the mTOR Pathway

Lin Wang  1 , Lin Zhang  2
Affiliations

Protein Kinase N2 Reduces Hydrogen Peroxide-inducedDamage and Apoptosis in PC12 Cells by AntiOxidative Stress and Activation of the mTOR Pathway

Lin Wang et al. Evid Based Complement Alternat Med. .

Abstract

Objective: To investigate the role and mechanism of protein kinase N2 (PKN2) in hydrogen peroxide (H2O2)-induced injury of PC12 cells.

Method: s. PC12 cells were transfected with lentivirus to knock down or overexpress PKN2 and then were treated with 300 μM H2O2 to establish a cell model of oxidative stress injury. The cell viability of PC12 cells in each group was determined by the CCK-8 method. Biochemical assays were used to measure reactive oxygen species (ROS), malondialdehyde (MDA) levels, and superoxide dismutase (SOD) activity. Western blot was used to detect the protein expressions of PKN2, caspase-3, cleaved-caspase-3, PARP, cleaved-PARP, p-mTOR, and mTOR in PC12 cells in each group.

Results: H2O2 treatment could significantly reduce PC12 cell viability and promote cell apoptosis and oxidative stress. PKN2 overexpression inhibited H2O2-induced apoptosis and oxidation damage by increasing PC12 cell viability, SOD activity, and p-mTOR protein expression, reducing intracellular ROS and MDA levels, and cleaved-caspase-3 and cleaved-PARP protein expression.

Conclusion: PKN2 overexpression can alleviate H2O2-induced oxidative stress injury and apoptosis in PC12 cells by activating the mTOR pathway.

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Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
PKN2 overexpression has a protective effect on H2O2-induced PC12 cells. (a/b) PKN2 protein expression in PC12 cells after transfection were detected by western blot. (c) Effects of PKN2 knockdown or overexpression on H2O2-induced PC12 cell viability. ∗∗P < 0.01vs. control, siNC + H2O2, and vector + H2O2.
Figure 2
Figure 2
PKN2 overexpression reduces H2O2-induced oxidative damage in PC12 cells. (a–c) Quantitative analysis of intracellular levels of ROS, (a) MDA, and (b) relative activity of SOD in each group. ∗∗P < 0.01vs. control, siNC + H2O2, and vector + H2O2.
Figure 3
Figure 3
PKN2 overexpression prevents H2O2-induced apoptosis of PC12 cells. (a) The protein expression levels of PARP, cleaved PARP, caspase-3, and cleaved caspase-3 in cells of each group were detected by western blot. (b–e) Image-Pro Plus software to analyze the gray values of PARP, cleaved PARP, caspase-3, and cleaved-caspase-3 proteins in each group of cells ∗∗P < 0.01vs. control, siNC + H2O2, and vector + H2O2.
Figure 4
Figure 4
PKN2 overexpression inhibits H2O2-induced apoptosis in PC12 cells by activating the mTOR pathway. (a–b) Western blot detection of mTOR and p-mTOR protein expression and p-mTOR/mTOR ratio in cells of each group. ∗∗P < 0.01vs. control, siNC + H2O2, and vector + H2O2.
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