Molecular Mechanism Of MicroRNA-146a Regulating HIV Infected Macrophages

Objective:In HIV / AIDS patients,macrophages are the "repository" of HIV,and are important antigen-presenting cells in the natural immune response.It has been reported that some microRNAs can participate in the regulation of macrophage antiviral innate immune process.The aim of this study is to explore the regulatory role of miR-146a in the innate immune response of macrophages against HIV by establishing a cell model of macrophages infected with HIV in vitro.Methods : Firstly,macrophages were isolated and cultured,and HIV virus was obtained by plasmid transfection.Then,the macrophage model of HIV infection was established,and the infection was verified by q-PCR.After HIV infected macrophages,cells were collected,total RNA was extracted by Trizol,and miR-146a with the largest change in expression was screened by q-PCR.The mimics and inhibitors of miR-146a were synthesized and transfected into macrophages respectively to overexpress or inhibit the expression of miR-146a.Then the regulatory effect of miR-146a on HIV infected macrophages and its effect on natural immunity were detected by q-PCR and ELISA.q-PCR and Western blot were used to explore the regulatory mechanism of miR-146a in the process of HIV infected macrophages.Resμlts : After successfully establishing the cell model of human primary macrophages infected with HIV in vitro,we found that the expression of miR-146a increased significantly after macrophages infected with HIV by q-PCR.After miR-146a was transfected into macrophages,and the macrophages were overexpressed / inhibited by miR-146a,it was found that miR-146a could promote HIV infected macrophages by q-PCR.In the overexpression of miR-146a group,it was confirmed by q-PCR and ELISA that miR-146a decreased IFN-α secretion of macrophages after HIV infection,and the expression of ISGs was also down regulated.In the inhibition of miR-146a group,the trend was just the opposite.It was confirmed by q-PCR and Western blot that miR-146a promoted the expression of SOCS-1 protein in HIV infected macrophages.Conclusions : After HIV infected macrophages,the expression of miR-146a increased.MiR-146a inhibited the secretion of IFN-α and the expression of ISGs by up regulating the expression of SOCS-1 protein,and finally promoted the expression of HIV infected macrophages.