The Research On The Role Of NF-κB In The Process Of Idiopathic Orbital Inflammatory Pseudotumor

Idiopathic Orbital Inflammatory Pseudotumor (IOIP) is a common orbit diseaseand a damage caused by inflammatory cells infiltration and fibrosis. It is a softtissue tumor. As the etiology of IOIP is not clear, diagnosis and treatment remains asignificant challenge in ophthalmology. To study the pathogenesis of the diseasebecomes extremely urgent.Nuclear factor-κB (NF-κB) is a transcription factor which widely exists in theeukaryotic cell. It can combine with gene promotor or enhancer of a variety cells topromote transcription and expression. NF-κB plays most important and evolutionarilyconserved role in the immune system, regulating the expression of inducers andeffectors at many points in the expansive networks that define responses to pathogens.In this study, we collect clinical IOIP samples and study the role of NF-κB inpathogenesis of IOIP. We finished the following work:1．Set up sample library. We collect clinical eye diseases about200cases. Thereare about70cases of IOIP samples, including blood and tissue. The blood iscentrifuged to separate plasma and blood cell. Next, we extract DNA from blood. Partof the tissue is made to paraffin section and HE staining. We classify the sampleaccording to the type of tissue cell. Other tissues are used extract protein, RNA andculture primary cell. The establishment of Sample Library can lay solid foundation ofIOIP study and have important significance.2．Use IHC, ICC, IFC, WB and real-time RT-PCR to test the expression ofNF-κB/p65, p50, IκB，IκBζ and p-p65in IOIP. And we use of microarray studiesNF-κB-related gene expression and suspension of NF-κB-chip system-relatedcytokines. The results showed:(1)p65is significantly up-regulated in nucleus andcytoplasm. Some samples with p65nucleus positive indicate p65plays a role oftranscription and activation. IκBζ is strong positive expressed and NFKBIZ gene issignificantly up-regulated. p-p65is strong positive expressed in nucleus and p50isstrong positive expressed in cytoplasm and nucleus.(2) NF-κB related proteinsexpressed in many cell types, mainly lymphocytes, fibroblasts, epithelial cells andother inflammatory cells.(3) The mainly NF-κB dimers is p65-p50in IOIP, also existp65-p65and p50-p50.(4) We cultured IOIP primary cell of IOIP tissues and foundmany fibrosis. Results of ICC of IOIP primary cells indicate that p65，IκBα，IκBζ are strongly positive in nucleus and cytoplasm. So, the fibrosis mechanism of IOIP haverelationship with the activation of NF-κB signaling pathway.(5) The microarrayresults of this study indicated that part of NF-κB-related genes were upregulated,NF-κB signaling pathway may be activated. Liquid suspension microarray resultsshowed that the MIF values of IOIP group were significantly different to the controlgroup.In conclusion, our study indicated that NF-κB signaling pathway is activated inIOIP and it has relationship with pathogenesis of IOIP. This study can provide theorybasis for diagnosis and treatment of IOIP.