Study On The Effect And Mechanism Of Anti-SIV/HIV Virus Of Icariin And Anhydroicaritin In Vitro

ObjectiveThis study is designed to explore:?The effects of Icariin and its metabolite Anhydroicaritin on the inhibition of Monkey Immunodeficiency Virus and Human immunodeficiency virus in Vitro.?The target of action for Icariin and its metabolite Anhydroicaritin inhibited Human immunodeficiency virus in vitro.?Observe the synergistic effects of between Icariin,Anhydroicaritin and Integrase inhibitors,respectively.Method?Icariin and Anhydroicaritin were tested for their cytotoxicity using CCK-8.?The inhibitory effects of Icariin and Anhydroicaritin on SIV /HIV-1 replication were assessed by Cytopathic effect(CPE)and real-time quantitative PCR(RT-PCR).?The mechanism of Icariin and Anhydroicaritin on anti-HIV-1 was explored by means of Computer Assisted Technology-Molecular Docking Method.?Further Icariin and Anhydroicaritin were detected for their ability to inhibit HIV-1 reverse transcriptase(RT)(HIV-1 RT)and HIV-1 protease(HIV-1 PR)by ELISA method and fluorescence method respectively.?Synergistic interaction was evaluated between Icariin,Anhydroicaritin and Integrase inhibitors by observation of cytopathic effect(CPE)and real-time quantitative RT-PCR.Results?The maximum non-toxic concentration of Icariin and and its metabolite Anhydroicaritin is 50? g//mL,16? g/mL,respectively.?For SIVmac251,in the cells,the viral load of Icariin(50,25,12.5? g/mL)and Anhydroicaritin(16,8,4? g/mL)are significantly lower than that of the virus control group(P<0.01,P <0.05).In the supernatant,the viral load of Icariin(50,25? g/mL)and Anhydroicaritin(16,8? g/mL)are also significantly lower than that of the virus control group(P <0.01,P <0.05).?For HIV-1,in the cells,the viral load of Icariin(50,25? g/mL)and Anhydroicaritin(16,8? g/mL)are significantly lower than that of the virus control group(P<0.01,P<0.05).In the supernatant,the viral load of Icariin(50,25? g/mL)and Anhydroicaritin(16? g/mL)are also significantly lower than that of the virus control group(P<0.01,P<0.05).?The result of molecular docking indicated that the anti-HIV activity is mediated through inhibition of HIV-1 reverse transcriptase and HIV-1 protease activity.?The results of ELISA and Fluorescence showed that Icariin(500? g/mL)and Anhydroicaritin(50? g/mL)could significantly inhibit the activity of HIV-1 RT,the inhibition rates are 83% and 89 %,respectively.And the positive drug EFV inhibition rate is 90%.Icariin(2mg/mL)and Anhydroicaritin(2mg/mL)significantly inhibited HIV-1 protease activity,the inhibition rates are 83% and 79%,respectively,and the positive inhibition rate is 85%.?In the aspect of synergistic interaction,Icariin,Anhydroicaritin and integrase inhibitor Raltegravir on HIV-1 inhibition are higher than the single drug group.Conclusions?Icariin and its metabolite Anhydroicaritin inhibited the replicatio n of SIV/HIV-1 virus,respectively.?The mechanism of Icariin and Anhydro icaritin on anti-HIV-1 are HIV-1 reverse transcriptase(HIV-1 RT)and HIV-1 protease(HIV-1 PR),respectively.?Icariin,Anhydroicaritin and integr ase inhibitor Raltegravir on HIV-1 inhibition are higher than the single drug group.