The Expression And Significance Of TLR4,MyD88mRNA In Cholesteatoma

BackgroudCholesteatoma is a common clinical disease, it has the continued growth of the tumor-likecapacity and the ability of bone destruction，so ,the cholesteatoma has been paid widen attentionby domestic and foreign experts. Although the causes of middle ear cholesteatoma and itsepithelial source there are still a lot of controversy, but the basic consensus is that the formationof cholesteatoma and the relevant body for the prevention of chronic inflammation, abnormalimmune response. Abnormal immune response is the important factor to cause of cholesteatomacontinued development and bone destruction. Toll-like receptor （toll.1ikereceptors TLRs） areinnate immunity pattern recognition receptors （pattern recognition receptor, PRR）, one of thespecial structure of the identification of pathogenic microorganisms and their cell wall products,the pathogen associated molecular patterns （pathogen associated molecularpatterns, PAMPs） therapid activation of the innate immune system ... Now M Szczepanski, WSzyfter found thatTLR （TLR-2, TLR-3, of TLR-4） express in the cholesteatoma microenvironment, suggesting thatTLR4 may be involved in the pathogenesis of cholesteatoma. In this study, a SYBR Greenreal-time PCR（SYBR-RT-PCR） are adopted, to observe the expression of TLR4 in theinflammatory response in varying degrees of middle ear cholesteatoma and its downstreamtransduction molecules MyD88, to further investigate TLR4 signal transduction pathways in thepathogenesis of cholesteatoma. In-depth understanding of cholesteatoma pathogenesis, and toprovide new ideas for the clinical treatment of cholesteatoma.MethodsSelection of middle ear cholesteatoma tissue specimens from 35 patients, both from theShanxi Medical University Second Department of Otolaryngology - head and neck surgeryinpatients 35 cases （ male 13, female22 cases, aged 16- 62 years old, average37.12 years old ）,ear pus history from 1 months to30 years, mean 5.14 years;8 cases of ossicular without destroyer,damaging1in 8 cases, destruction of2in 13 cases,6 cases of destruction3. During intraoperativespecimens according to the microscopic veiw of subepithelial stromal inflammation degree,sampling into inflammatory and non-inflammatory group, in which inflammatory group13,non-inflammatory group12. The control group for the normal external ear canal skin tissue in 10cases, also taken from the middle ear surgery. All of the above specimens under a microscope, carefully remove cholesteatoma scales and other necrotic tissue, the refrigerator at minus 80degrees Celsius, as used in RT-PCR detection. All patients underwent temporal bone highresolution CT examination, postoperative pathologic biopsy were confirmed to have acquiredsecondary to cholesteatoma. Using real-time fluorescence quantitative PCR （ Real-time PCR ）for direct detection of inflammatory group, non-inflammatory group, normal control group, theexpression of TLR4、MyD88mRNA.Result1. Real time-PCR method to directly detect the three groups of experimental samples ofTLR4, the inflammatory group TLR4 average 2-^ΔΔCT0..689（0.200,2.050）, non-inflammatorygroup TLR4 average 2-^ΔΔCT0.291 （0.020,0.990）, and normal controlgroup of TLR4, an averageof 2-^ΔΔCT0.163 （0.001,0.990）.2. Real time-PCR method for direct detection of the three groups of experimental samplesMyD88mRNA expression: inflammation group the MyD88 average 2-^ΔΔCTvalue 0.777（0.320,1.84）; non-inflammatory group the MyD88 an average of 2-^ΔΔCTvalue of 0.320（0.01,0.990） the normal control group MyD88 average 2-^ΔΔCTvalue of 0.224 （0.002,0.990）.3. Inflammatory group of TLR4, MyD88mRNA expression levels than normal externalauditory canal group increased by 4 times and 3 times, and the difference was statisticallysignificant （p <0.05）; inflammatory group of TLR4, MyD88mRNA expression levels comparedwith non-inflammatory group were increased by 2 and 1 times, and the difference wasstatistically significant （p <0.05）、4.We will do TLR4and MyD88 Spearman rank correlation analysis, the correlationcoefficient r = 0.955, P < 0.05. This is show that in cholesteatoma otitis media pathogenesis,TLR4mRNA and MyD88 are positively related.Conclusion1. Middle ear cholesteatoma tissue in the TLR4mRNA, MyD88mrNA expression2. Middle ear, especially the inflammatory reaction of cholesteatoma, the expression ofTLR4 and its downstream signal transduction molecules change, suggesting that the incidence ofotitis media with cholesteatoma is indeed closely related with the degree of inflammation.3. The inflammatory reaction of the cholesteatoma epithelium tissue expression of TLR4raised its downstream transduction molecules MyD88 enhanced expression of both a positivecorrelation. We conclude that of TLR4-of MyD88 signaling pathway may be an importantmechanism of cholesteatoma.