The Expression And Signifance Of Tlr-2,tlr-4 And Nf-kb In Middle Ear Cholesteatoma

ObjectiveCholesteatoma of the middle ear is a common type in chronic otitis media,characterized by of hyperproliferative keratunizing squamous epithelium in the middle ear cavity and destruction of adjacent bone.It has adverse effect on ear structures,resulting in osteolysis and bone resporption,threatening the lives of patients.Its formation and pathogenesis are not fully understood.This study aimed to analyze the expression of Nuclear factor-kappa B(NF-kB),toll-like receptor 4 (TLR-4),toll-like receptor 2(TLR-2) in middle ear cholesteatoma,and the relevance between TLR-2,TLR-4 and the NF-kB,and to investigate the TLR-2,TLR-4,NF-kB in human middle ear cholesteatoma in the process of inflammatory response role.Methods46 case of cholesteatoma specimens were collected at time of sugery,and 15 cases of the normal skin of external acoustic meatus were the control group.All of the speciemens were fixed immediately and made into paraffin section. Immunohistochemical staining was used to detect the expression of TLR-2,TLR-4 and NF-kB in the cholesteatoma and the skin of external acoustic meatus.The computer image quantitative analysis was used to examin the expression TLR-2,TLR-4 and NF-kB.Results1.The expression of TLR-2 in cholesteatoma and the skin of external acoustic meatus: The positive immunostaining of TLR-2 appeared all the layers over subepithelial stromal cells and the cytoplasm,and TLR-2 protein was localize mostly in the cytoplasm,include basal layer,stratum spinosum,stratum granulosum,stratum corneum,but the distribution and frequency of the positive cells varied,nucleus-free coloring.The skin of external acoustic meatus showed only at the picchi bottom.2.The expression of TLR-4 in cholesteatoma and the skin of external acoustic meatus:The positive immunostaining of TLR-4 appeared all the layers,TLR-4 protein was localized mostly in the cytoplasm,and it showed the tendency of gradual increasing from basal layer to corneous layer,nucleus-free coloring,cholesteatoma subepithelial connective tissue can be seen scattered positive cells,mainly composed of fibroblasts,monocytes-macrophages,plasma cells,a few lymphocytes.The skin of external acoustic meatus showed only at the picchi bottom.3.The expression of NF-kB in cholesteatoma and the skin of external acoustic meatus:NF-kB in 46 cases of cholesteatoma epithelium,NF-kB-positive cells distributed in the all epithelial layer,different cholesteatoma intensity ranging from the positive expression,the majority of specimens showed strong positive expression;Each example specimen has the cytoblastema coloration,partial specimen nucleus are positive staining.Cholesteatoma subepithelial inflammatory cells and vascular epithelial cells can also be found scattered in the positive expression.15 cases of normal skin of the external auditory canal see full-thickness skin of the weak positive expression of cytoplasmic expression of both.4.The gray scale value of TLR-2,TLR-4 and NF-kB(1)The mean gray scale value of TLR-2 was 138.61±6.20 in cholesteatoma and 95.64±6.20 in the skin of external acoustic meatus,and there was statistical significance between the groups(t=20.73,P＜0.05).(2)The mean gray scale value of TLR-4 was 140.87±5.23 in cholesteatoma and 100.45±2.88 in the skin of external acoustic meatus,and there was statistical significance between the groups(t=28.48,P＜0.05).(3)The mean gray scale value of NF-kB was 155.25±11.72 in cholesteatoma and 95.65±8.12 in the skin of external acoustic meatus,and there was statistical significance between the groups(t=17.96,P＜0.05)5.After the spearman rank correlation analysis,in the cholesteatoma organization between TLR-2 and NF-kB has the positive correlational(r=0.316, p＜0.05);Between TLR-4 and NF-kB has the positive correlational(r=0.389,p＜0.01).Conclusions1.TLR-2,TLR-4,NF-kB-mediated chronic inflammatory response and immune response,this study showed that TLR-2,TLR-4,NF-kB was highly expressed in cholesteatoma,suggesting that in chronic inflammatory response and the immune response may play a certain function.2.TLR-2 participated in inflammatory of cholesteatoma pathogen micro-environment to stimulate the surface of PAMPs by intracellular signal transduction pathways,activation of nuclear factor NF-kB,NF-kB through a wide range of the regulation of gene expression,regulation and control of many inflammatory mediators and cytokines the expression of inflammatory response involved in the body.There was a correlation between TLR-2 and NF-kB,they may have been synergistic in the forming cholesteatoma.3.TLR-4 participated in inflammatory of cholesteatoma pathogen micro-environment to stimulate the surface of PAMPs by intracellular signal transduction pathways,activation of nuclear factor NF-kB,NF-kB through a wide range of the regulation of gene expression,regulation and control of many inflammatory mediators and cytokines the expression of inflammatory response involved in the body.There was a correlation between TLR-4and NF-kB,they may have been synergistic in the forming cholesteatoma.