| Human immunodeficiency virus（HIV）is a kind of retroviruse which mainly attack human immune system cells,and cause the immune system lose its function,finally generate AIDS（Acquired Immunodeficiency Syndrom）.AIDS has been widespread in the world and has been a serious threaten to human life and health.Therefore,it is necessary to develop new drugs for different targets of HIV.At present,there is no report on anti-HIV-1 activity of Tibetan Medicine,and its related chemical and pharmacological research is scarce and the mechanism is still not clear,In this study,we evaluated the anti-HIV-1 virus activity and mechanism of 8 kinds of Tibetan medicine by in vitro pharmacodynamic experiment.The antiviral activity of 8 kinds of Tibetan medicine ethanol extracts were evaluated by CCK-8 and in vitro infection model,Fallopia aubertii,Anemone vitifolia Buch,Saxifraga melanocentra Franch,Comarum salesovianum and Chamaenerion angustifolium extracts had good antiviral activity and the half maximal inhibitory concentation(IC50)were 6.47±0.78、11.97±1.09、11.70±0.79、6.53±1.01、4.00±0.88μg/mL,Surface plasmon resonance（SPR）showed that Anemone vitifolia Buch and Saxifraga melanocentra Franch with Integrase,Anemone vitifolia Buch with Vpu,Anemone vitifolia Buch and Saxifraga melanocentra Franch with Vif,Fallopia aubertii with gp41 had good combination.The in vitro experimental of inhibiting activity to recombinant Reverse Transcriptase,Integrase and Protease showed that Fallopia aubertii,Anemone vitifolia Buch and Saxifraga melanocentra Franch had significant inhibitory effect on so7d-IN activity,IC500 were 27.08、36.77、42.93μg/mL.The study of Comarum salesovianum and Chamaenerion angustifolium were Carried out on the basis of previous research,The samples were extracted with petroleum ether,ethyl acetate,n-butanol and water,and detecting their anti-virus activity in vitro,The IC500 of Comarum salesovianum ethyl acetate,n-butanol and water extracts were 4.13±0.46、2.30±0.62、2.97±0.53μg/mL,Chamaenerion angustifolium petroleum ether,ethyl acetate,n-butanol and water extracts were 1.48±0.13、3.09±0.57、6.5±0.82、1.14±0.02μg/mL.HIV-1 R3A wild virus infection inhibition experiment showed that chamaenerion angustifolium water extract has good inhibition to the wild virus,and the IC500 detected by P24antigen detection and luciferase assay was 16.09、10.56μg/mL,SPR indicited that Comarum salesovianum and Chamaenerion angustifolium with Integrase had good combination,in addition and Chamaenerion angustifolium had good combination with Vif.The in vitro experimental of inhibiting activity to recombinant Reverse Transcriptase,Integrase and Protease showed that Comarum salesovianum N-butanol extract and Chamaenerion angustifolium water extract had significant inhibitory effect on Sso7d-IN activity and the inhibition rate were 95.87%、84.54%,Chamaenerion angustifolium water extract had significant inhibitory effect on Protease and the inhibition rate was 79.71%.in addition,The Western blot experiments showed that Chamaenerion angustifolium water extract could down the expression of Vif protein.The result showedthatFallopia aubertii,Anemone vitifolia Buch,Saxifraga melanocentra Franch,Comarum salesovianum and Chamaenerion angustifolium extracts could potently inhibit the HIV-1 activity in vitro,Chamaenerion angustifolium water extract had good inhibitory effect and light Cytotoxic to HIV-1 Pseudovirus and wild strains,and its mechanism could be related to the inhibition of integrase 3’processing and down regulation of Vif protein expression.This study will provide experimental basis and foundation for the research of Tibetan medicine and the development of new antiviral drugs. |
