【作者】 李咏玲；

【导师】 徐志文；

【作者基本信息】 广西医科大学 ， 耳鼻咽喉—头颈外科， 2008， 硕士

【摘要】 目的:探讨脂多糖、维生素D受体、基质金属蛋白酶-9蛋白在中耳胆脂瘤中的含量及表达的情况和它们之间的关系。方法:标本来自在广西医科大学第一附属医院耳鼻咽喉科行手术治疗的29例胆脂瘤型中耳炎患者,其中18例来自这些胆脂瘤患者的正常外耳道皮肤作为对照组。所有胆脂瘤病例全部经病理确诊。标本分两部分收集,一部分用无热原冻存管于-80℃下保存用作LPS含量测定,采用鲎变性细胞溶解物测定法(LAL法);另一部分经甲醛固定、石蜡包埋,用作鼠抗人VDR与鼠抗人MMP-9蛋白的免疫组化,检测其表达情况。结果:结果发现,LPS在胆脂瘤组织中含量比在正常外耳道皮肤组织中表达高(均为取对数后数值):胆脂瘤组中位数(Median,M)=0.7390,四分位数间距(interquartile range,IQR)=0.6203;外耳道组M=-0.2538,IQR=1.1692(P＜0.05)。在两种类型的胆脂瘤组织中,广泛型组M=0.8403,IQR=0.5254;局限型组M=0.4048,IQR=0.6139(P＜0.05),广泛型的LPS含量比局限型的高。VDR蛋白在外耳道上皮呈阳性表达,而在胆脂瘤上皮呈中、低表达。VDR蛋白阳性定位于细胞核,呈棕黄色至褐色颗粒状。在胆脂瘤组,VDR阳性率为93.1%,与外耳道皮肤相比,具有显著差异(P＜0.05)。MMP-9蛋白在胆脂瘤上皮中基本呈阳性表达,而在外耳道皮肤上皮中呈中、低表达或不表达。MMP-9蛋白阳性定位于细胞质,呈棕黄色颗粒状。在胆脂瘤标本中,MMP-9阳性细胞率为79.3%,与外耳道皮肤相比,具有显著差异(P＜0.05);在两种类型的胆脂瘤中,MMP-9和VDR的表达差异无统计学意义(P＞0.05)。经Spearman检验LPS、VDR、MMP-9之间无相关关系。结论:1、LPS在胆脂瘤中的含量高于外耳道皮肤,且胆脂瘤广泛型组比局限型组含量增高,提示LPS与胆脂瘤的形成及其骨质破坏有关。2、VDR在外耳道皮肤的表达高于胆脂瘤上皮,提示VDR与胆脂瘤的发生有关。3、MMP-9在胆脂瘤上皮中的表达高于外耳道皮肤,提示MMP-9与胆脂瘤的发生有关。4、LPS、VDR、MMP-9三者无相关关系,提示三者在中耳胆脂瘤发生中的作用尚无明确的联系。更多还原

【Abstract】 Objective: To investigate the expression and concentration of lipopolysaccharide, vitamin D receptor and matrix metalloproteinase-9 in middle ear cholesteatoma and discuse their relations.Methods: 29 cases of middle ear cholesteatoma tissue we studied were come from the patients who were operated in First Affliliated Hospital of Guangxi Medical University, 18 cases of external auditory canal tissue were acquired from those patients at the same time, and all of them had been verified by pathology. Tissues were stored into 2 parts, ones were stored without any additive in pyrogen-free tubes at -80 C until further processing, the other ones were formalin-fixed, paraffin-embedded for immunohistochemical test. The concentrations of LPS were detected by limulus amebocyte lysate assay (LAL-assay) , and expression of VDR and MMP-9 protein in formalin-fixed, paraffin-embedded tissues were detected by immunohistochemical method.Results: The result showed that the concentrations of LPS in cholesteatoma are higher than in external auditory canal tissues (the numbers are logarithmed): Group of Cholesteatoma: M=0. 7390, IQR=0. 6203; Group of external auditory canal tissues : M=-0.2538 , IQR=1. 1692 (P<0.05) . Between the two-type groups: Group of intensive type: M=0. 8403, IQR=0. 5254; Group of localized type: M=0. 4048, IQR=0. 6139, the concentrations of LPS were higher in extensive cholesteatoma in comparison with localized cholesteatoma (P<0.05) . VDR was located at karyon, taking on yellow brown pellet, could be detected in all external auditory canal epithelium but there were less positive cells in middle ear cholesteatoma. In cholesteatoma epithelium samples, VDR were 93.1%. MMP-9 was located at cytoplasmic, taking on yellow brown pellet, could be detected in almost middle ear cholesteatoma epithelium but there were less positive cells in external auditory canal epithelium. In cholesteatoma epithelium samples, MMP-9 were 79.3%. Compared with external auditory canal epithelium, the expression of MMP-9 was higher in middle ear cholesteatoma epithelium (P<0.05), but was lower in expression of VDR(P<0.05). They didn’t have significant difference in two types of cholesteatoma epithelium(P>0.05). LPS, MMP-9 and VDR weren’t significant correlated by Spearman test.Conclusion:1 The concentrations of LPS were higher in cholesteatoma compared with external auditory canal, and were higher in extensive types compared with localized ones, which hints LPS was responsible for middle ear cholesteatoma and the bone erosion of cholesteatoma.2 The expression of VDR was higher in external auditory canal compared with cholesteatoma, which hints VDR was responsible for middle ear cholesteatoma.3 The expression of MMP-9 was higher in cholesteatoma compared with external auditory canal, which hints MMP-9 was responsible for middle ear cholesteatoma.4 There’s no correlation among LPS, VDR and MMP-9, which hints the relationships among the three were unkown.更多还原