Protocatechuic aldehyde mitigates hydrogen peroxide-triggered PC12 cell damage by down-regulating MEG3

Figures & data

Figure 1. H₂O₂ caused PC12 cells damage. (A) PC12 cells were received the diverse dosages of H₂O₂ (0–400 µM) management, CCK-8 experiment was conducted to detect cell viability. The dosage of 200 µM H₂O₂ was selected and utilized to manage PC12 cells. CCK-8, flow cytometry and western blot experiments were implemented to examine (B) cell viability, (C) apoptosis and (D,E) Bax, Pro/Cleaved-Caspase-3 and Pro/Cleaved-Caspase-9 expression. *p < .05, **p < .01, ***p < .001.

Figure 2. PA assuaged H₂O₂-triggered PC12 cell damage. (A) PC12 cells received the different dosages of PA (0–2 µM) administration, CCK-8 experiment was executed to test cell viability. (B) H₂O₂ (200 µM) and PA (0.1, 0.5 and 1 µM) were exploited to co-manage PC12 cells, CCK-8 experiment was carried out again to examine cell viability. H₂O₂ (200 µM) and PA (1 µM) were utilized to dispose PC12 cells, flow cytometry and western blot were conducted to evaluate (C) apoptosis and (D,E) Bax, Pro/Cleaved-Caspase-3 and Pro/Cleaved-Caspase-9 expression. *p < .05, **p < .01, ***p < .001.

Figure 3. PA relieved H₂O₂-triggered PC12 cell autophagy. H₂O₂ (200 µM) and PA (1 µM) were utilized to dispose PC12 cells, western blot was implemented to test (A,B) protein levels of cell autophagy-associated factors (Beclin-1, p62 and LC3-II/LC3-I) in PC12 cells. **p < .01, ***p < .001.

Figure 4. PA caused MEG3 down-regulation in H₂O₂-damaged PC12 cells. (A) PA (1 µM) was utilized to manage PC12 cells, RT-qPCR was performed to detect MEG3 expression in these disposed PC12 cells. (B) H₂O₂ (200 µM) and PA (1 µM) were exploited to manage PC12 cells, RT-qPCR was conducted to determine MEG3 expression in these disposed PC12 cells. (C) Expression vectors of pc-MEG3 and pcDNA3.1 were applied for PC12 cells transient transfection, and then RT-qPCR was performed again to determine MEG3 expression in these transfected PC12 cells. *p < .05, **p < .01.

Figure 5. PA allayed H₂O₂-triggered PC12 cells damage through prohibiting MEG3 expression. PC12 cells received pc-MEG3 and pcDNA3.1 transfection and then were managed with H₂O₂ (200 µM) and PA (1 µM), CCK-8, flow cytometry and western blot were executed to test (A) cell viability, (B) apoptosis, (C,D) Bax, Pro/Cleaved-Caspase-3 and Pro/Cleaved-Caspase-9 expression and (E,F) cell autophagy-associated factors (Beclin-1, p62 and LC3-II/LC3-I) expression. *p < .05, **p < .01, ***p < .001.

Figure 6. PA expedited Wnt/β-catenin and PTEN/PI3K/AKT activations in H₂O₂-managed PC12 cells through adjusting MEG3 expression. PC12 cells were received pc-MEG3 and pcDNA3.1 transfection meanwhile were managed with H₂O₂ (200 µM) and PA (1 µM), western blot was conducted to estimate the protein levels of (A,B) Wnt3a, β-catenin and Wnt5a as well as (C,D) PTEN, p/t-PI3K and p/t-AKT in above-managed PC12 cells. *p < .05, **p < .01, ***p < .001.