Cepharanthine and Curcumin inhibited mitochondrial apoptosis induced by PCV2

Figures

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  Fig. 1. Dose-response curves of all tested compounds in cytotoxicity assay. 8 of gradient dilutions of each compound was prepared by 2-fold serial dilution and incubated with cells for 60 h. Dose-response curve of the tested compounds used in PK-15 shown the more cytopathic effect were observed with the increased concentration of compounds

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  Fig. 2. Anti-PCV2 activities of Paeonol, Cepharanthine or Curcumin detected by qPCR and Western blot. The anti-PCV2 effect of compounds was evaluated by qPCR and Western blot. Cells were incubated with 104.4 TCID50 of PCV2 for 2 h, then tested compounds were added respectively and cultured for 48 h. Cells were collected and relative assay perform. a The expression of the Cap gene was detected by qPCR; b The expression of the Cap protein was detected by Western blot. GraphPad Prism 5.0 was used for the statistical analysis. All data were presented in Mean ± SEM. Different letters to indicate significant differences between groups (p<0.05)

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  Fig. 3. Inhibition of PCV2-induced cell apoptosis by Cepharanthine or Curcumin. a and b After cells were infected with 104.4 TCID50 of PCV2 for 2 h, cells were treated with Cepharanthine, Curcumin or Ribavirin for 48 h. The apoptosis rate was evaluated by Annexin V/PI staining followed by flow cytometry. The data on the graph indicated the total number of cells at the late apoptosis of the right upper quadrant and the early apoptosis of the right lower quadrant. The results indicated apoptosis rates were significantly decreased in the treated group with Cepharanthine, Curcumin or Ribavirin Compared to the PCV2-infected group except the group of 0.005 mg/mL Curcumin (P < 0.05)

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  Fig. 4. Inhibition of PCV2-induced cell apoptosis by Cepharanthine or Curcumin via the mitochondria pathway. a and b The changes of MMP in each group after being staining with JC-1 kit were detected by flow cytometry. R2 represents the value of MMP. The results indicated MMP values were significantly decreased in the treated group with Cepharanthine, Curcumin or Ribavirin Compared to the PCV2-infected group (P < 0.05)

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  Fig. 5. The mechanism of inhibition of PCV2-induced mitochondrial apoptosis by Cepharanthine or Curcumin. After cells were infected with 104.4 TCID50 of PCV2 for 2 h, cells were treated with Cepharanthine, Curcumin or Ribavirin for 48 h. a-h The expression of the apoptin were analyzed by Western blot and grayscale analysis. h, i The results of flow cytometry showed that ROS values were significantly decreased in the treated group with Cepharanthine at 0.003 and Curcumin at 0.02, 0.01 and 0.005 mg/mL Compared to the PCV2-infected group (P < 0.05)

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  Fig. 6. The PCV2-induced Mitochondrial apoptosis was mainly remitted by Cepharanthine and Curcumin. The “red arrow” represents the promoting effect and the red “T-shape” indicates the inhibiting effect. The PCV2 replication is inhibited by Cepharanthine and Curcumin, selected from 13 natural compounds, through mitochondrial apoptosis pathway

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  Fig. 7. The chemical structure of 13 natural compounds