The Expression and Signifance of TLR-2、TLR-4 and NF-kB in Middle Ear Cholesteatoma
|Course||Department of Otolaryngology Head and Neck Surgery|
|Keywords||Middle ear cholesteatoma TLR-2 TLR-4 NF-kB|
Objective cholesteatoma otitis media is a common type of chronic otitis media, Its main feature is the performance of the middle ear cavity to proliferate keratinizing squamous epithelium and adjacent bone destruction to which the main bone destruction characteristics, can lead to severe intracranial foreign complications, threatening the lives of patients. The current study has not been fully elucidated the exact mechanism of the formation of cholesteatoma. The purpose of this study is to analyze Toll-like receptors (toll-like receptor TLR-2) and Toll-like receptor 4 (toll-like receptor 4 and TLR-4), a nuclear transcription factor (Nuclear factor-kappa B, NF -kB) expression in cholesteatoma and distribution, and analysis of the correlation between the TLR-2, TLR-4 and NF-kB, explore the TLR-2 and TLR-4, NF-kB in the middle ear Cholesterol The role of tumor inflammatory process. Materials and methods In this study, collected fresh specimens of 46 cases of patients with otitis media with cholesteatoma surgery, including 15 cases of cholesteatoma patients in normal skin at the same time take the external auditory canal, as normal controls. Samples were taken in 10% formalin fixed paraffin sections. Immunohistochemistry (SP method) was used to detect the TLR-2, TLR-4 and NF-kB expression in middle ear cholesteatoma and external auditory canal skin, and a computer image analysis system for TLR-2, TLR-4 and NF -kB staining results of quantitative analysis. Results 1.TLR-2 expression in cholesteatoma tissues and normal external ear canal skin TLR-2 in 46 cases of cholesteatoma epithelial tissue showed a moderate positive or strong positive reaction showed brown particles clouds in the epithelial each layer and on in the the subcutaneous stromal cell cytoplasm, including basal cell layer, spinous layer, granular layer, the stratum corneum, but the distribution and frequency of positive cells varies nucleus coloring. Only 15 cases of normal external ear canal skin tissue expression in the basal underlying. 2.TLR-4 expression in cholesteatoma tissues and normal external ear canal skin TLR-4 in 46 cases of cholesteatoma epithelial tissue showed a moderate positive or strong positive reaction showed brown particles in clouds in the epithelial layers of cells cytoplasm, showing from the basal layer of the cuticle staining gradually increased trend nucleus coloring. Subcutaneous visible on cholesteatoma fibroblasts, monocytes, macrophages, and plasma cells, a small number of lymphocytes and other scattered positive cells. Only on basal underlying weak expression in 15 cases of normal external ear canal skin tissue. Positive expression intensity of expression of NF-kB 3.NF-kB in cholesteatoma and normal external ear canal skin in 46 cases of cholesteatoma epithelial tissue, NF-kB positive cells located in the epithelial layers of different cholesteatoma range, the majority of specimens were strongly positive expression; nucleus positive staining of both cytoplasmic staining specimens, some specimens. Cholesteatoma on the the subcutaneous inflammatory cells and vascular endothelial cells also can be seen scattered in the positive expression. 15 cases of normal external auditory canal skin tissue to see a weak expression of the whole layer of the epidermis, are cytoplasmic expression. Computer image analysis system to detect TLR-2 and TLR-4 and TLR-NF-kB gray values ??(1) 2 in 46 cases of cholesteatoma epithelial tissue grayscale value of 138.61 ± 6.20 in the 15 cases of normal external auditory canal skin in the average gray value of 95.64 ± 6.20, the between difference was statistically significant (t = 20.73, p <0.05). (2) TLR-4 in the 46 cases of cholesteatoma epithelial tissue grayscale value of 140.87 ± 5.23, the 15 normal ear canal skin mean gray value of 100.45 ± 2.88, significant difference between the two (t = 28.48, p <0.05). (3) NF-kB in 46 cases of cholesteatoma epithelial tissue grayscale value is 155.25 ± 11.72, the 15 normal ear canal skin mean gray value of 95.65 ± 8.12, significant difference between the two (t = 17.96, p <0.05). 5 correlation analysis via spearman level, cholesteatoma between TLR-2 and NF-kB positive correlation (r = 0.316, p <0.05); TLR-4 and NF-kB exists a positive correlation between ( r = 0.389, p <0.01). IV. Conclusions 1.TLR-2, TLR-4, NF-kB-mediated chronic inflammation and immune responses, the study results show that TLR-2 and TLR-4, the high expression of NF-kB in cholesteatoma , suggesting that chronic inflammatory reaction in the middle ear cholesteatoma play a certain role. The 2.TLR-2 feelings cholesteatoma the inflammatory microenvironment pathogen surface PAMPs stimulation via intracellular signaling pathways, activation of the nuclear factor NF-kB, NF-kB through a variety of gene expression regulation, regulation of many inflammatory mediators and cells expression of the factor involved in the body's inflammatory response. The results of this study show a positive relationship between TLR-2 and NF-kB, suggesting that TLR-2 and NF-kB may be a synergistic effect in the inflammatory reaction in the middle ear cholesteatoma. The 3.TLR-4 feelings cholesteatoma the inflammatory microenvironment pathogen surface PAMPs stimulation via intracellular signaling pathways, activation of the nuclear factor NF-kB, NF-kB through a variety of regulation of gene expression regulation of numerous inflammatory mediators and cell expression of the factor involved in the body's inflammatory response. The results of this study show a positive relationship between TLR-4 and NF-kB, suggesting that TLR-4 and NF-kB possible synergies in the inflammatory reaction in the middle ear cholesteatoma.