김현준 Song, Jung Whan 2012-08 12660 https://aurora.ajou.ac.kr/handle/2018.oak/17998 학위논문(박사)아주대학교 일반대학원 :의학과,2012. 8 TABLE OF CONTENTS ABSTRACT i TABLE OF CONTENTS ii LIST OF FIGURES iv LIST OF TABLE v Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 3 A. Cell culture and LTA treatment 3 B. RT-PCR 3 C. Real-time PCR 4 D. Enzyme-linked immunosorbent assay (ELISA) 5 E. Gelatin zymography 5 F. Western blot analysis 6 G. MEK1/2 inhibitor treatment 7 H. Statistical analysis 7 Ⅲ. RESULTS 8 A. Expression of MMPs and TIMPs mRNAs in HMEE cells after treatment with LTA 8 B. Real-time PCR of MMP-9 8 C. Secretion of MMP-9 8 D. Activation of MMP-9 enzyme (gelatinase B) 12 E. Western blot analysis 12 F. MEK1/2 inhibitor treatment 12 Ⅳ. DISCUSSION 16 Ⅴ. CONCLUSION 19 REFERENCES 20 국문요약 24 eng The Graduate School, Ajou University 아주대학교 논문은 저작권에 의해 보호받습니다. 인체중이점막세포 (human middle ear epithelial cells)에서 포도상구균 (Staphylococcus aureus)의 lipoteichoicacid에 의한 MMP-9의 발현 Jung Whan Song Thesis 아주대학교 일반대학원 Jung Whan Song 일반대학원 의학과 2012. 8 Master http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012660 matrix metalloproteinase middle ear otitis media Lipoteichoic acid from Staphylococcus aureus induced expression of MMP-9 in human middle ear epithelial cells Change in matrix metalloproteinases (MMPs) and regulation of their tissue inhibitors of metalloproteinases (TIMPs) could play certain role in the pathogenesis of otitis media. This study was designed to evaluate the modulation of MMPs and TIMPs in middle ear epithelium by lipoteichoic acid (LTA) isolated from Staphylococcus aureus.Human middle ear epithelial cells (HMEE) were treated with LTA. Changes of MMP and TIMP and signal transduction were examined by PCR, ELISA, zymography and western blot. LTA isolated from S. aureus increased MMP-9 mRNA expression and secretion in HMEEC, whereas no effect on the expressions of MMP-1, 2, 3, 7 and TIMP-1, 2 was observed. Only the extracellular signal-regulated kinases (ERK) 1/2 of mitogen-activated protein kinase (MAPK) pathway was activated by LTA. And the MEK1/2 inhibitor prevented the expression of MMP-9 by LTA.LTA increased the activity of MMP-9, not TIMPs in middle ear epithelia, suggesting that disturbed balance between MMP-9 and TIMPs could play an active role in LTA-induced otitis media.