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Resveratrol exhibits a strong cytotoxic activity in cultured cells and has an antiviral action against polyomavirus: potential clinical use.

Berardi V, Ricci F, Castelli M, Galati G, Risuleo G - J. Exp. Clin. Cancer Res. (2009)

Bottom Line: This compound has a strong bioactivity and its cytoprotective action has been demonstrated, however at high concentrations the drug exhibits also an effective anti-proliferative action.We recently showed its ability to abolish the effects of oxidative stress in cultured cells.The results presented here show an evident toxic activity of the drug at high concentrations, on the other hand at sub-cytotoxic concentrations, resveratrol can effectively inhibit the synthesis of polyomavirus DNA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Genetica e Biologia Molecolare, Sapienza Università di Roma, Roma, Italy. valerio.berardi@uniroma1.it

ABSTRACT

Background: Resveratrol is a non flavonoid polyphenol compound present in many plants and fruits and, at especially high concentrations, in the grape berries of Vitis vinifera. This compound has a strong bioactivity and its cytoprotective action has been demonstrated, however at high concentrations the drug exhibits also an effective anti-proliferative action. We recently showed its ability to abolish the effects of oxidative stress in cultured cells. In this work we assayed the bioactivity of resveratrol as antiproliferative and antiviral drug in cultured fibroblasts. Studies by other Authors showed that this natural compound inhibits the proliferation of different viruses such as herpes simplex, varicella-zoster and influenza A. The results presented here show an evident toxic activity of the drug at high concentrations, on the other hand at sub-cytotoxic concentrations, resveratrol can effectively inhibit the synthesis of polyomavirus DNA. A possible interpretation is that, due to the damage caused by resveratrol to the plasma membrane, the transfer of the virus from the endoplasmic reticulum to the nucleus, may be hindered thus inhibiting the production of viral DNA.

Methods: The mouse fibroblast line 3T6 and the human tumor line HL60 were used throughout the work. Cell viability and vital cell count were assessed respectively, by the MTT assay and Trypan Blue staining. Cytotoxic properties and evaluation of viral DNA production by agarose gel electrophoresis were performed according to standard protocols.

Results: Our results show a clear dose dependent both cytotoxic and antiviral effect of resveratrol respectively at high and low concentrations. The cytotoxic action is exerted towards a stabilized cell-line (3T6) as well as a tumor-line (HL60). Furthermore the antiviral action is evident after the phase of virion entry, therefore data suggest that the drug acts during the synthesis of the viral progeny DNA.

Conclusion: Resveratrol is cytotoxic and inhibits, in a dose dependent fashion, the synthesis of polyomavirus DNA in the infected cell. Furthermore, this inhibition is observed at non cytotoxic concentrations of the drug. Our data imply that cyto-toxicity may be attributed to the membrane damage caused by the drug and that the transfer of polyomavirus from the endoplasmic reticulum to the cytoplasm may be hindered. In conclusion, the cytotoxic and antiviral properties of resveratrol make it a potential candidate for the clinical control of proliferative as well as viral pathologies.

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Related in: MedlinePlus

Decrease of viral DNA as a function of the duration of the exposure to resveratrol. Left panel: Progeny viral DNA (form I) is indicated by the arrow. In this case, the culture medium was changed to fresh drug-free medium at the following times post-infection. The incubation was continued for 24 hours. Lane 1: Mock infected cells; Lane 2: Untreated control cells; Lane 3 through 6: 4, 8, 12 and 16 hours, respectively; Lane 7: The medium was not changed and infection was carried permanently in the presence of RV (20 μM). Right panel: Quantification of the fluorescence bands reported in the left panel. The yield of the viral DNA is normalized to the amount obtained in untreated control cells (Bar 1). Withdrawal of RV is reported in the legend to left panel of this figure.
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Figure 3: Decrease of viral DNA as a function of the duration of the exposure to resveratrol. Left panel: Progeny viral DNA (form I) is indicated by the arrow. In this case, the culture medium was changed to fresh drug-free medium at the following times post-infection. The incubation was continued for 24 hours. Lane 1: Mock infected cells; Lane 2: Untreated control cells; Lane 3 through 6: 4, 8, 12 and 16 hours, respectively; Lane 7: The medium was not changed and infection was carried permanently in the presence of RV (20 μM). Right panel: Quantification of the fluorescence bands reported in the left panel. The yield of the viral DNA is normalized to the amount obtained in untreated control cells (Bar 1). Withdrawal of RV is reported in the legend to left panel of this figure.

Mentions: To assess whether the continuous presence of RV is necessary to inhibit the viral replication we removed the drug at different time points after the viral penetration into the cell (Figure 3). Therefore, the infection was carried out in 20 μM RV but the culture medium was changed to a drug-free fresh medium after different times of treatment and the incubation was continued for 24 hours. Results show that removal of RV after four hour incubation has little or no effect on the yield of viral progeny DNA (lane 2). The drug must be present for the whole infection time to be effective and to cause the complete inhibition of the viral replication (lanes 6 and 7).


Resveratrol exhibits a strong cytotoxic activity in cultured cells and has an antiviral action against polyomavirus: potential clinical use.

Berardi V, Ricci F, Castelli M, Galati G, Risuleo G - J. Exp. Clin. Cancer Res. (2009)

Decrease of viral DNA as a function of the duration of the exposure to resveratrol. Left panel: Progeny viral DNA (form I) is indicated by the arrow. In this case, the culture medium was changed to fresh drug-free medium at the following times post-infection. The incubation was continued for 24 hours. Lane 1: Mock infected cells; Lane 2: Untreated control cells; Lane 3 through 6: 4, 8, 12 and 16 hours, respectively; Lane 7: The medium was not changed and infection was carried permanently in the presence of RV (20 μM). Right panel: Quantification of the fluorescence bands reported in the left panel. The yield of the viral DNA is normalized to the amount obtained in untreated control cells (Bar 1). Withdrawal of RV is reported in the legend to left panel of this figure.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2710315&req=5

Figure 3: Decrease of viral DNA as a function of the duration of the exposure to resveratrol. Left panel: Progeny viral DNA (form I) is indicated by the arrow. In this case, the culture medium was changed to fresh drug-free medium at the following times post-infection. The incubation was continued for 24 hours. Lane 1: Mock infected cells; Lane 2: Untreated control cells; Lane 3 through 6: 4, 8, 12 and 16 hours, respectively; Lane 7: The medium was not changed and infection was carried permanently in the presence of RV (20 μM). Right panel: Quantification of the fluorescence bands reported in the left panel. The yield of the viral DNA is normalized to the amount obtained in untreated control cells (Bar 1). Withdrawal of RV is reported in the legend to left panel of this figure.
Mentions: To assess whether the continuous presence of RV is necessary to inhibit the viral replication we removed the drug at different time points after the viral penetration into the cell (Figure 3). Therefore, the infection was carried out in 20 μM RV but the culture medium was changed to a drug-free fresh medium after different times of treatment and the incubation was continued for 24 hours. Results show that removal of RV after four hour incubation has little or no effect on the yield of viral progeny DNA (lane 2). The drug must be present for the whole infection time to be effective and to cause the complete inhibition of the viral replication (lanes 6 and 7).

Bottom Line: This compound has a strong bioactivity and its cytoprotective action has been demonstrated, however at high concentrations the drug exhibits also an effective anti-proliferative action.We recently showed its ability to abolish the effects of oxidative stress in cultured cells.The results presented here show an evident toxic activity of the drug at high concentrations, on the other hand at sub-cytotoxic concentrations, resveratrol can effectively inhibit the synthesis of polyomavirus DNA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Dipartimento di Genetica e Biologia Molecolare, Sapienza Università di Roma, Roma, Italy. valerio.berardi@uniroma1.it

ABSTRACT

Background: Resveratrol is a non flavonoid polyphenol compound present in many plants and fruits and, at especially high concentrations, in the grape berries of Vitis vinifera. This compound has a strong bioactivity and its cytoprotective action has been demonstrated, however at high concentrations the drug exhibits also an effective anti-proliferative action. We recently showed its ability to abolish the effects of oxidative stress in cultured cells. In this work we assayed the bioactivity of resveratrol as antiproliferative and antiviral drug in cultured fibroblasts. Studies by other Authors showed that this natural compound inhibits the proliferation of different viruses such as herpes simplex, varicella-zoster and influenza A. The results presented here show an evident toxic activity of the drug at high concentrations, on the other hand at sub-cytotoxic concentrations, resveratrol can effectively inhibit the synthesis of polyomavirus DNA. A possible interpretation is that, due to the damage caused by resveratrol to the plasma membrane, the transfer of the virus from the endoplasmic reticulum to the nucleus, may be hindered thus inhibiting the production of viral DNA.

Methods: The mouse fibroblast line 3T6 and the human tumor line HL60 were used throughout the work. Cell viability and vital cell count were assessed respectively, by the MTT assay and Trypan Blue staining. Cytotoxic properties and evaluation of viral DNA production by agarose gel electrophoresis were performed according to standard protocols.

Results: Our results show a clear dose dependent both cytotoxic and antiviral effect of resveratrol respectively at high and low concentrations. The cytotoxic action is exerted towards a stabilized cell-line (3T6) as well as a tumor-line (HL60). Furthermore the antiviral action is evident after the phase of virion entry, therefore data suggest that the drug acts during the synthesis of the viral progeny DNA.

Conclusion: Resveratrol is cytotoxic and inhibits, in a dose dependent fashion, the synthesis of polyomavirus DNA in the infected cell. Furthermore, this inhibition is observed at non cytotoxic concentrations of the drug. Our data imply that cyto-toxicity may be attributed to the membrane damage caused by the drug and that the transfer of polyomavirus from the endoplasmic reticulum to the cytoplasm may be hindered. In conclusion, the cytotoxic and antiviral properties of resveratrol make it a potential candidate for the clinical control of proliferative as well as viral pathologies.

Show MeSH
Related in: MedlinePlus