Research On The Mechanism Of HIV-1 Gp120 Mediated Central Nervous System Injury By α7nAchR

BackgroundIn addition to AIDS,HIV-1 infection can also cause nervous system disease hand.The combined use of antiretroviral therapy(cRT)has made the mortality rate of AIDS drop sharply.However,the prevalence of hand is increasing,and there is no specific drug for hand,which seriously affects the quality of life of the patients.Gp120 is one of the main neurotoxins that lead to hand.Gp120 can activate microglia,release inflammatory factors,cause CNS inflammation,and promote apoptosis of neurons,thus damaging the nervous system and causing the occurrence of hand.But the specific mechanism is not clear.ObjectiveThe aim of this study is to study the mechanism of HIV-1 gp120 mediated apoptosis by A7R in vivo and vitro,to determine the role of A7R in the development of hand,to clarify the effect of MEM hydrochloride on neurocognitive impairment,and to construct a mouse model of gp120 transgenic and A7R gene knockout,which provides a basis for further study of the mechanism of gp120 induced nerve injury.Methods1.Construction of a transgenic mouse model of A7R gene knockout HIV-1 gp120Gp120 TG mice(gp120+)and A7R knockout homozygous mice(α7R-/-)were selected and labeled FO.First,heterozygotes F1 were obtained(α7R+/-/gp 120+或α7R+//gp120-).After genotyping,the genotypes of α7R+/-/gp120+were screened out and mated freely to obtain F2 generation mice.Finally,it was screened out from F2 mice that the mice with α7R-/-/gp120+ genotype were used as the successful individuals of double transgenic model.2.Treatment of microglia and neurons by gp120 and a7r inhibitorsThe expression of A7R,p53,Bax,Bcl-2 and caspase 3 were detected by Western blot.ROS production and apoptosis of BV2 cells were detected by flow cytometry.3.Intraperitoneal injection of MEM into gp120 Tg miceWild type and gp120 TG mice were divided into four groups,each group of 6:WT,WT+MEM,gp120 Tg,gp120 Tg+MEM.The mice in WT+mem and gp120 TG+MEM groups were injected with MEM every two days at 20mg/kg dose,and PBS was injected into WT and gp120 Tg groups for 90 days.Morris water maze was performed after intervention,and the expression of brain tissue related proteins was detected by immunohistochemistry and Western blot.Results1.Both gene and protein showed that the model of gp120 transgenic mice was successfully knocked out by A7R gene.2.The indirect treatment of gp120 increased the apoptosis of neurons,and decreased the apoptosis after inhibition of A7R.3.Gp120 increased the expression of A7R,ROS production,p53,Bax and caspase 3,bcl-2 expression,MLA pretreatment,and p53,Bax and caspase 3,bcl-2.4.The expression of p53,Bax,caspase 3 and Bcl-2 in the brain tissue of gp120 TG mice increased,bcl-2 decreased,and the expression of p53,Bax and caspase 3 decreased after MEM injection,and Bcl-2 increased5.The learning and memory ability of gp120 TG mice was significantly reduced,and the cognitive impairment of gp120 TG mice was improved by injecting mem.Conclusion1.The model of G120 transgenic mice was successfully constructed by A7R gene knockout.2.Gp120 was mediated by A7R and apoptosis was induced indirectly by ROSp53-bax-caspase 3 pathway.3.MEM can improve the neurocognitive impairment caused by gp120.