Transcriptome Analysis Of The Spinal Cord Of HIV-induced Neuropathic Pain Mouse And Identification Of Related Functional Genes

Human immunodeficiency virus(HIV)infection is one of the most dramatic public health problems in the world today.Highly active antiretroviral therapy makes HIV infection tend to be a chronic disease,so improving the quality of life of patients has become a new focus of HIV related research.HIV-induced neuropathic pain(HNP)severely reduces the quality of life of patients,leading to pain,insomnia,and depression.Currently,there is no drug to control it.The fundamental reason is that the molecular mechanism of HNP is not clear.Therefore,it is of great significance to strengthen the study on the pathological mechanism of HNP.In this study,‘transcriptome analysis of the spinal cord of HNP mouse and identification of related functional genes’ will provide new insights into the study of the pathological mechanism of HNP.Firstly,the HNP mouse model was established by intrathecal injection of HIV envelope glycoprotein gp120.Transcriptome sequencing analysis was performed on the spinal cord of HNP mice,and the results are as follows:(1)There were 1682,430 and 413 Differentially expressed genes(DEGs)at 2 h(POD 1/12),1 d(POD 1)and 14 d(POD 14)after modeling,respectively.(2)Bioinformatics analysis showed the enrichment of inflammatory response,immune response,oxidative damage,PI3K-AKT signaling pathway,NF-κB signaling pathway and Toll-like receptor signaling pathway in POD 1/12 group;Immune response,inflammatory response,synaptic vesicle pathway,NF-κB signaling,gamma-aminobutyric acid(GABA)metabolism and m TOR signaling pathway enrichment in POD 1 group;Enrichment of IL-5 signaling,TGF-β signaling,mitochondrial damage,downregulation of ion channel activity,endocytosis,MAPK signaling pathway and neurotrophic protein signaling pathway in POD 14 group.Secondly,the key genes and candidate genes that mediated the different stages of HNP were screened by bioinformatics method:(1)In POD 1/12 group,19 key genes including Ptgs2,Tnf and Nos1 and 15 candidate genes including Tlr2 and Cxcl10 were found.(2)10 key genes,such as Ptgs2 and P2rx7,and 19 candidate genes,such as Socs3 and Src,were found in POD 1 group.(3)In POD 14 group,8 key genes such as Syt2 and Kcna2 and 19 candidate genes such as Shank1 and Gan were found.Ptgs2,Nos1,Tlr2,Socs3 and Syt2 were further verified by qPCR and/or Western blotting.Activation of PI3K-AKT and MAPK signaling pathways was verified by Western blotting.Finally,the role of candidate gene Tlr2 in HNP was selected for preliminary exploration.Immunofluorescence colocalization showed that TLR2 was mainly expressed and upregulated in HNP mouse spinal microglia.Intrathecal injection of TLR2 inhibitor o-Vanillin reduced the mechanical allodynia in HNP mice,while inhibiting the activation of PI3K-AKT signaling pathway,reducing the expression of inflammation-related factors PTGS2 and TNF,and reducing the expression of oxidative damage-related protein NOS1.In summary,this study shows that in the initial stage of HNP(2h and 1d after modeling),immunity,inflammation and oxidative damage are involved in this process.The main key genes are Tlr2,Ptgs2,Tnf,Aif1,Nos1 and Socs3,the main key signaling pathways is the PI3K-AKT pathway;In the maintenance phase of HNP(14 days after modeling),mitochondrial damage and destruction of ion homeostasis are involved in this process.The main key gene is Syt2,and the main key signaling pathway is the MAPK pathway.gp120 is involved in the acute initiation of HNP by activating microglia and increasing inflammatory response and oxidative damage through the TLR2/PI3K/ AKT signaling axis.