Antiviral activity of shikonin ester derivative PMM-034 against enterovirus 71 in vitro

Figures

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  Figure 1.. Chemical structure of PMM-034.

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  Figure 2.. Cytotoxicity of shikonin ester derivative PMM-034 on rhabdomyosarcoma cell viability. PMM-034 was serially diluted to 0, 2.5, 5, 10, 20, 50, 100, 200, and 400 μg/mL in DMEM containing 2.5% FBS. Subsequently, the cytotoxicity of PMM-034 on RD cells was determined by the WST-1 assay. Data are reported as means ± standard deviation (SD) from at least three independent experiments. The red dot indicates the PMM-034 concentration under 50% cell survival rate.

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  Figure 3.. Dose- and time-dependent effects of PMM-034 on human enterovirus 71 (EV71) replication in rhabdomyosarcoma (RD) cells. A, RD cells were infected with EV71 for 1 h, then treated with PMM-034 or DMSO for 23 h. Infected cells and culture supernatants were collected 48 h post-infection. B, RD cells were infected with EV71 for 1 h, and 10 μg/mL PMM-034 was added at 0, 2, 4, 8, 12, 16, and 24 h after infection. Virus titers were measured by the plaque reduction assay. Data are reported as means±SD from at least 3 independent experiments. *P<0.05; **P<0.01, Student’s t-test.

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  Figure 4.. Effects of PMM-034 on human enterovirus 71 (EV71/VP1) mRNA and protein expressions in rhabdomyosarcoma (RD) cells. RD cells were infected with EV71 for 1 h and then treated with 10 μg/mL PMM-034 or DMSO for 23 h. A, The EV71/VP1 mRNA level was detected by real-time RT-PCR. GAPDH was used as the internal control. Data are reported as means±SD of at least 3 independent experiments, **P<0.01, Student’s t-test. B, Expression levels of EV71/VP1 protein were detected by western blotting. β-actin was used as the internal control.

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  Figure 5.. Effects of PMM-034 on the mRNA levels of pro-inflammatory cytokines and phospho-p65 protein expression in rhabdomyosarcoma (RD) cells. RD cells were infected with human enterovirus 71 (EV71) for 1 h, then treated with 10 μg/mL PMM-034 or DMSO for 23 h. A, mRNA levels of IL-1β, IL-6, IL-8, and TNF-α were detected by real-time RT-PCR. GAPDH was used as the internal control. Data are reported as means±SD of at least 3 independent experiments. **P<0.01, Student’s t-test. B, Phospho-p65 protein expression was detected by western blotting. β-actin was used as the internal control.