Abstract
Objective
To investigate the effect and potential mechanism of dihydromyricetin (Dmy) on H9C2 cell proliferation, apoptosis, and autophagy.
Methods
H9C2 cells were randomly divided into 7 groups, namely control, model, EV (empty pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro vector), IV (circHIPK3 interference), Dmy (50 µ mol/L), Dmy+IV, and Dmy+EV groups. Cell proliferation and apoptosis were detected by cell counting kit-8 assay and flow cytometry, respectivley. Western blot was used to evaluate the levels of light chain 3 II/I (LC3II/I), phospho-phosphoinositide 3-kinase (p-PI3K), protein kinase B (p-AKT), and phospho-mammalian target of rapamycin (p-mTOR). The level of circHIPK3 was determined using reverse transcriptase polymerase chain reaction. Electron microscopy was used to observe autophagosomes in H9C2 cells.
Results
Compared to H9C2 cells, the expression of circHIPK in H9C2 hypoxia model cells increased significantly (P<0.05). Compared to the control group, the cell apoptosis and autophagosomes increased, cell proliferation rate decreased significantly, and the expression of LC3 II/I significantly increased (all P<0.05). Compared to the model group, the rate of apoptosis and autophagosomes in IV, Dmy, and Dmy+IV group decreased, the cell proliferation rate increased, and the expression of LC3 II/I decreased significantly (all P<0.05). Compared to the control group, the expressions of p-PI3K, p-AKT, and p-mTOR in the model group significantly reduced (P<0.05), whereas after treatment with Dmy and sh-circHIPK3, the above situation was reversed (P<0.05).
Conclusion
Dmy plays a protective role in H9C2 cells by inhibiting circHIPK expression and cell apoptosis and autophagy, and the mechanism may be related to PI3K/AKT/mTOR pathway.
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Data Availability Statement
The data used to support the findings of this study can be obtained from the corresponding author upon request.
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Zhang ZY, Liu C, and Wang PX drafted the manuscript. Han YW and Zhang YW reviewed and modified the manuscript. Hao ML, Song ZX, and Zhang XY revised the manuscript. All authors contributed to the article and approved the submitted version.
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Supported by Science Foundation of Education Department of Shaanxi Provincial Government (No. 19JK0890), Natural Science Foundation of Xizang (Tibet) Autonomous Region (No. XZ202001ZR0089G), Major Training Project of Xizang Minzu University (Nos. 18MDZ03 and 20MDT03), and Funded Project of Young Scholar Cultivation Program of Xizang Minzu University (No. 21MDX04)
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Zhang, Zy., Liu, C., Wang, Px. et al. Dihydromyricetin Alleviates H9C2 Cell Apoptosis and Autophagy by Regulating CircHIPK3 Expression and PI3K/AKT/mTOR Pathway. Chin. J. Integr. Med. 29, 434–440 (2023). https://doi.org/10.1007/s11655-022-3687-4
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DOI: https://doi.org/10.1007/s11655-022-3687-4