二氢青蒿素(dihydroartemisinin,D H A)是青蒿素的一种衍生物,在多种肿瘤中表现出明显的抗肿瘤活性,但其具体机制不详。本文探讨了D H A对肝癌细胞的毒性作用机制。利用C C K-8试剂检测D H A对肝癌细胞株活力的影响,通过荧光探针染色及...二氢青蒿素(dihydroartemisinin,D H A)是青蒿素的一种衍生物,在多种肿瘤中表现出明显的抗肿瘤活性,但其具体机制不详。本文探讨了D H A对肝癌细胞的毒性作用机制。利用C C K-8试剂检测D H A对肝癌细胞株活力的影响,通过荧光探针染色及流式细胞术分析细胞内R 0 S及脂质过氧化物水平的变化;通过谷胱甘肽测定试剂盒检测细胞内还原型谷胱甘肽含量的变化,并通过免疫印迹分析D H A作用下细胞内铁死亡通路蛋白质中G P X 4的变化。结果发现,D H A能显著抑制SMMC-7721及H uh-7细胞活力,其半数抑制浓度分别为23.74 p m o l/L及26.92 jjumol/L。在35 pm ol/L D H A处理下,SMMC-7721及H uh-7细胞内R 0 S分别升高2.6倍和2.1倍,月旨质过氧化物升高2.3倍和1.7倍。D H A可诱导细胞内G SH含量下降,并能下调铁死亡相关蛋白质G P X 4蛋白水平。通过利用小分子抑制剂进行功能恢复实验发现,R 0 S抑制剂、铁螯合剂及铁死亡抑制剂都可不同程度恢复D H A引起的细胞活力下降。进一步检测发现,铁死亡抑制剂可抑制D H A诱导的脂质过氧化,并恢复G SH含量及G P X 4蛋白水平。结果表明,在肝癌细胞中,D H A可通过诱导细胞发生铁死亡抑制肝癌细胞生长。展开更多
Rabies is caused by infection of rabies virus(RABV)and remains a serious threat to the global public health.Except for the requirement for cold chain and high cost of human rabies immune globulin,no small molecule dru...Rabies is caused by infection of rabies virus(RABV)and remains a serious threat to the global public health.Except for the requirement for cold chain and high cost of human rabies immune globulin,no small molecule drugs are currently available for clinical treatment of rabies.So,it is of great importance to identify novel compounds that can effectively inhibit RABV infection.Artesunate(ART)and dihydroartemisinin(DHA),two derivatives of artemisinin,are widely used for treatment of malaria in adults and children,showing high safety.In this study,we found that both ART and DHA were able to inhibit RABV replication in host cells at a low concentration(0.1μmol/L).The antiviral effects of ART and DHA were independent of viral strains and cell lines.Pre-treatment with ART or DHA for 2 h in vitro did not affect the viral replication in host cells,implying that ART and DHA neither reduced the viability of RABV directly nor inhibited the binding and entrance of the virus to host cells.Further studies revealed that ART and DHA inhibited RABV genomic RNA synthesis and viral gene transcription.Treatment with ART or DHA(5 mg/kg)by intramuscular injection improved,to some extent,the survival rate of RABV-challenged mice.Combination treatment with derivatives of artemisinin and mannitol significantly improved the survival rate of RABV-challenged mice.The results suggest that ART and DHA have a great potential to be explored as new anti-rabies agents for treatment of rabies.展开更多
文摘二氢青蒿素(dihydroartemisinin,D H A)是青蒿素的一种衍生物,在多种肿瘤中表现出明显的抗肿瘤活性,但其具体机制不详。本文探讨了D H A对肝癌细胞的毒性作用机制。利用C C K-8试剂检测D H A对肝癌细胞株活力的影响,通过荧光探针染色及流式细胞术分析细胞内R 0 S及脂质过氧化物水平的变化;通过谷胱甘肽测定试剂盒检测细胞内还原型谷胱甘肽含量的变化,并通过免疫印迹分析D H A作用下细胞内铁死亡通路蛋白质中G P X 4的变化。结果发现,D H A能显著抑制SMMC-7721及H uh-7细胞活力,其半数抑制浓度分别为23.74 p m o l/L及26.92 jjumol/L。在35 pm ol/L D H A处理下,SMMC-7721及H uh-7细胞内R 0 S分别升高2.6倍和2.1倍,月旨质过氧化物升高2.3倍和1.7倍。D H A可诱导细胞内G SH含量下降,并能下调铁死亡相关蛋白质G P X 4蛋白水平。通过利用小分子抑制剂进行功能恢复实验发现,R 0 S抑制剂、铁螯合剂及铁死亡抑制剂都可不同程度恢复D H A引起的细胞活力下降。进一步检测发现,铁死亡抑制剂可抑制D H A诱导的脂质过氧化,并恢复G SH含量及G P X 4蛋白水平。结果表明,在肝癌细胞中,D H A可通过诱导细胞发生铁死亡抑制肝癌细胞生长。
基金We thank Dr.Xianzhu Xia(Academy of Military Medical Sciences)for providing the CVS-11 virus.We thank Dr.Zhenfang Fu(Huazhong Agricultural University)for providing the CVS-24 virus.This work was supported by the National Key Research and Development Program of China(2016YFD0500400)National Natural Science Foundation of China(31772742)Natural Science Foundation of Guangdong(2015A03031103).
文摘Rabies is caused by infection of rabies virus(RABV)and remains a serious threat to the global public health.Except for the requirement for cold chain and high cost of human rabies immune globulin,no small molecule drugs are currently available for clinical treatment of rabies.So,it is of great importance to identify novel compounds that can effectively inhibit RABV infection.Artesunate(ART)and dihydroartemisinin(DHA),two derivatives of artemisinin,are widely used for treatment of malaria in adults and children,showing high safety.In this study,we found that both ART and DHA were able to inhibit RABV replication in host cells at a low concentration(0.1μmol/L).The antiviral effects of ART and DHA were independent of viral strains and cell lines.Pre-treatment with ART or DHA for 2 h in vitro did not affect the viral replication in host cells,implying that ART and DHA neither reduced the viability of RABV directly nor inhibited the binding and entrance of the virus to host cells.Further studies revealed that ART and DHA inhibited RABV genomic RNA synthesis and viral gene transcription.Treatment with ART or DHA(5 mg/kg)by intramuscular injection improved,to some extent,the survival rate of RABV-challenged mice.Combination treatment with derivatives of artemisinin and mannitol significantly improved the survival rate of RABV-challenged mice.The results suggest that ART and DHA have a great potential to be explored as new anti-rabies agents for treatment of rabies.
