The Research On HIV-1 Latency Reactivation By Extract Of Three Kinds Of Thymelaeaceae Plants

Although highly active antiretroviral therapy(HAART)can effectively inhibit the replication of human immunodeficiency virus(HIV)to an undetectable level,it still can not clear the virus completely.The main reason why AIDS can not be cured is due to latent infection of HIV in the body.“Shock and kill” is a new strategy in the treatment of HIV infection.In this study,HIV-1 latency reactivation model(J-Lat 11.1 cells)was adopted to evaulate the reactivation effects of 60% ethanol extract of Wikstroemia indica,100% ethanol extract of Daphne genkwa and 60% ethanol extract of Lowdaphne Stringbush Flower by flow cytometry.The percentage of green fluorescent protein(GFP)positive cells reflects the extent of HIV latent infection reactivation.The results showed that the effective dose range of the 60% ethanol extract of Wikstroemia indica,100% ethanol extract of Daphne genkwawas or 60% ethanol extract of Lowdaphne Stringbush Flower was about 0.33 ?g/m L-208.30 ?g/m L,0.08 ?g/mL-52.63 ?g/m L or 0.05 ?g/mL-166.70 ?g/mL respectively.The reactivation effects of ethanol extracts from the three kinds of thymelaeaceae plants appeared gradually from 4 h post-treatment.The reactivation effects of these three ethanol extracts decreased(p<0.05)when J-Lat 11.1 cells were pre-treated with PKC inhibitor(G? 6983 or G? 6850)or NF-?B inhibitor(PDTC or BAY 11-7082),while PKC inhibitor(G? 6976)did not affect the reactivation effect(p>0.05).The results suggested that PKC(??,?,? or ?)subtype and NF-?B pathway may be involved in HIV latency reactivation by ethanol extracts.A real-time quantitative detection method for EGFP-PKC/NF-?B nuclear translocation was established with live cell imaging technique,and the effect of ethanol extracts on nuclear translocation of EGFP-PKC/NF-?B was investigated.The results showed that 60% ethanol extract of Wikstroemia indica and 60% ethanol extract of Lowdaphne Stringbush Flower were able to promote EGFP-PKC-? nuclear translocation.60% ethanol extract of Wikstroemia indica,100% ethanol extract of Daphne genkwa and 60% ethanol extract of Lowdaphne Stringbush Flower can promote EGFP-PKC-? and EGFP-p65 nuclear translocation.PKC inhibitor(G? 6983)inhibited the nuclear translocation of EGFP-p65 by 60% ethanol extract of Wikstroemia indica and 100% ethanol extract of Daphne genkwa,and could not inhibit the nuclear translocation of EGFP-p65 by 60% ethanol extract of Lowdaphne Stringbush Flower.G? 6850 and G? 6976 did not affect the nuclear translocation of EGFP-p65 by these three kinds of ethanol extracts.These results suggested the nuclear translocation of NF-?B p65 by 60% ethanol extract of Wikstroemia indica or 100% ethanol extract of Daphne genkwa might be mediated by PKC-?/? subtype,and PKC might not participate in the nuclear translocation of NF-?B p65 by 60% ethanol extract of Lowdaphne Stringbush Flower.