Inhibitory activities of 3-O-β-chacotriosyl pentacyclic triterpenoids against the entry of H5 N1 influenza viruses in vitro / 中国药理学通报

Aim To study the inhibitory activities of potential new anti-influenza virus agents,3-O-β-chaco-triosyl pentacyclic triterpenoids against the entry of H5N1influenza viruses.Methods Three target com-pounds were designed and synthesized structurally re-lated to the lead compound 3-O-β-chacotriosyl dioscin derivative (1 )with inhibitory activities against H5N1 influenza viruses.The inhibitory activities of these tar-get compounds were tested at a cellular level pseudo vi-rus system targeting H5N1 influenza viruse entry.Re-sults All the compounds 1 a,1 b and 1 c showed po-tent inhibitory activities against the entry of A/Thai-land/Kan353/2004 pseudo virus into the target cells, of which compound 1 b showed the best inhibitory activ-ity with an IC50 value of (1.25 ±0.22)μmol·L-1. Conclusion The SARs analysis of these compounds indicated that replacement of the aglycone moiety of compound 1 with pentacyclic triterpenoids could in-crease antiviral activity.Different types of pentacyclic triterpen as aglycone residue had the significant influ-ence on the inhibitory activity (1 b >1 c >1 a),sug-gesting ursane type of triterpenes was superior to the two other kinds of triterpenes as aglycone residue.

Influenza virus H1N1 induced apoptosis of mouse astrocytes and the effect on protein expression

OBJECTIVE@#To investigate the effects of influenza A virus H1N1 infection on the proliferation and apoptosis of mouse astrocytes cells and its protein expression.@*METHODS@#After mouse astrocytes was infected with purified influenza A virus H1N1 in vitro, viral integration and replication status of the cells were detected by RT-PCR assay, cell proliferation and apoptosis was determined by MTT method and flow cytometry, respectively. Associated protein expression was detected by Western blotting.@*RESULTS@#Agarose gel electrophoresis showed H1N1 virus can infect astrocytes and can be copied. MTT staining showed H1N1 virus infection can inhibit the proliferation of mouse astrocytes, which makes cell viability decreased significantly. Flow cytometry showed that the proportion of Annein V staining positive vascular endothelial cells in the influenza A virus group was significantly higher than that in the control group. Western blot analysis showed after 24 h and 32 h of infection, there were cells caspase-3 protein and the expression of its active form (lysed caspase-3 protein) increased. The proportion of Bax/Bcl-2 also increased.@*CONCLUSIONS@#Influenza A virus can infect human vascular endothelial cells and proliferation and it can induce apoptosis of endothelial cells.