The iron chelators can be utilized in target cells to improve 5-aminolaevulinic acid （ALA）-based photodynamic therapy （PDT）. The purpose of this study is to compare the effect of two kinds of iron chelators, desfe...The iron chelators can be utilized in target cells to improve 5-aminolaevulinic acid （ALA）-based photodynamic therapy （PDT）. The purpose of this study is to compare the effect of two kinds of iron chelators, desferrioxamine （DFO） and ethylenediaminetetraacetic acid （EDTA） on the enhancement of ALA-PDT. HaCat cells were cultured in medium containing 2.0 mmol/L of ALA and 0.5 mmol/L of DFO or EDTA. After 3-h incubation in the dark, the concentration of cellular pro-toporphyrin Ⅸ （PpⅨ） was detected by high performance liquid chromatography （HPLC）, and the fluorescence of PpⅨ was observed at 630 nm emission under confocal laser scanning microscope. For PDT, HaCat cells were irradiated using 632.8 nm laser, and the fractions of apoptotic and necrotic cells were flow cytometrically assayed. Related differences in morphology and ultrastructure of Ha-Cat cells were observed using optical microscope or transmission electron microscope. Compared to incubation with ALA alone, the addition of DFO or EDTA increased the concentration of cellular PpⅨ and the fluorescent density of PpⅨ, and also increased cell death ratio after PDT. PDT using ALA plus DFO produced the highest cellular PpⅨ level, greatest cell death ratio and most severe structural damage to the cells. It was concluded that both DFO and EDTA could enhance ALA-based PpⅨ production and PDT. Compared to the non-specific iron chelator of EDTA, the specific chelator, DFO, showed more potential for the enhancement.展开更多
The SARS-CoV-2 infection is spreading rapidly worldwide.Efficacious antiviral therapeutics against SARSCo V-2 is urgently needed.Here,we discovered that protoporphyrin IX(Pp IX)and verteporfin,two Food and Drug Admini...The SARS-CoV-2 infection is spreading rapidly worldwide.Efficacious antiviral therapeutics against SARSCo V-2 is urgently needed.Here,we discovered that protoporphyrin IX(Pp IX)and verteporfin,two Food and Drug Administration(FDA)-approved drugs,completely inhibited the cytopathic effect produced by SARS-CoV-2 infection at 1.25 lmol/L and 0.31 lmol/L,respectively,and their EC50 values of reduction of viral RNA were at nanomolar concentrations.The selectivity indices of Pp IX and verteporfin were 952.74 and 368.93,respectively,suggesting a broad margin of safety.Importantly,Pp IX and verteporfin prevented SARS-CoV-2 infection in mice adenovirally transduced with human angiotensin-converting enzyme 2(ACE2).The compounds,sharing a porphyrin ring structure,were shown to bind viral receptor ACE2 and interfere with the interaction between ACE2 and the receptor-binding domain of viral S protein.Our study suggests that Pp IX and verteporfin are potent antiviral agents against SARS-CoV-2 infection and sheds new light on developing novel chemoprophylaxis and chemotherapy against SARS-CoV-2.展开更多
文摘The iron chelators can be utilized in target cells to improve 5-aminolaevulinic acid （ALA）-based photodynamic therapy （PDT）. The purpose of this study is to compare the effect of two kinds of iron chelators, desferrioxamine （DFO） and ethylenediaminetetraacetic acid （EDTA） on the enhancement of ALA-PDT. HaCat cells were cultured in medium containing 2.0 mmol/L of ALA and 0.5 mmol/L of DFO or EDTA. After 3-h incubation in the dark, the concentration of cellular pro-toporphyrin Ⅸ （PpⅨ） was detected by high performance liquid chromatography （HPLC）, and the fluorescence of PpⅨ was observed at 630 nm emission under confocal laser scanning microscope. For PDT, HaCat cells were irradiated using 632.8 nm laser, and the fractions of apoptotic and necrotic cells were flow cytometrically assayed. Related differences in morphology and ultrastructure of Ha-Cat cells were observed using optical microscope or transmission electron microscope. Compared to incubation with ALA alone, the addition of DFO or EDTA increased the concentration of cellular PpⅨ and the fluorescent density of PpⅨ, and also increased cell death ratio after PDT. PDT using ALA plus DFO produced the highest cellular PpⅨ level, greatest cell death ratio and most severe structural damage to the cells. It was concluded that both DFO and EDTA could enhance ALA-based PpⅨ production and PDT. Compared to the non-specific iron chelator of EDTA, the specific chelator, DFO, showed more potential for the enhancement.
基金supported by the National Science and Technology Major Project(NSTMP)for the Prevention and Treatment of Infectious Diseases(2018ZX10734401,2018ZX10301208)the NSTMP for the Development of Novel Drugs(2019ZX09721001)the Project of Novel Coronavirus Research of Fudan University,China Postdoctoral Science Foundation(2020T130016ZX)。
文摘The SARS-CoV-2 infection is spreading rapidly worldwide.Efficacious antiviral therapeutics against SARSCo V-2 is urgently needed.Here,we discovered that protoporphyrin IX(Pp IX)and verteporfin,two Food and Drug Administration(FDA)-approved drugs,completely inhibited the cytopathic effect produced by SARS-CoV-2 infection at 1.25 lmol/L and 0.31 lmol/L,respectively,and their EC50 values of reduction of viral RNA were at nanomolar concentrations.The selectivity indices of Pp IX and verteporfin were 952.74 and 368.93,respectively,suggesting a broad margin of safety.Importantly,Pp IX and verteporfin prevented SARS-CoV-2 infection in mice adenovirally transduced with human angiotensin-converting enzyme 2(ACE2).The compounds,sharing a porphyrin ring structure,were shown to bind viral receptor ACE2 and interfere with the interaction between ACE2 and the receptor-binding domain of viral S protein.Our study suggests that Pp IX and verteporfin are potent antiviral agents against SARS-CoV-2 infection and sheds new light on developing novel chemoprophylaxis and chemotherapy against SARS-CoV-2.
