【作者】 洪艺云；

【导师】 叶胜难；

【作者基本信息】 福建医科大学 ， 耳鼻咽喉科学， 2010， 硕士

【摘要】 目的:采用CD105做为微血管标记物,并以Ki-67作为胆脂瘤增殖的指标研究胆脂瘤上皮下结缔组织微血管生成在胆脂瘤增殖中的作用以及MMP9和VEGF在诱导血管生成中的作用,期望有助于揭示胆脂瘤的发病机理,并为其预防和治疗提供理论和临床依据。方法:通过免疫组织化学染色方法(二步法)检测MMP9、VEGF、Ki-67和CD105在45例中耳胆脂瘤组织和10例正常外耳道皮肤中的表达;采用Image-Pro plus6.0图像分析软件读取胆脂瘤标本和外耳道正常皮肤标本MMP9、VEGF蛋白的平均光密度值;计数MVD来评价微血管生成。结果:1、中耳胆脂瘤组织中见新生的微血管较丰富、密集,且在形态上表现出异常的改变。正常外耳道皮肤组织中的微血管较少,稀疏,没有明显形态学上的改变。45例中耳胆脂瘤组织中的平均MVD为28.3±7.7,10例正常外耳道皮肤平均MVD为6.1±2.5,两组间差异有统计学意义(t=8.905,p<0.001)。2、中耳胆脂瘤组织中Ki-67蛋白的阳性表达率显著高于正常外耳道皮肤(χ2=7.305,P<0.01);随着胆脂瘤上皮下结缔组织微血管密度(MVD)的增加,其Ki-67阳性表达率也相应地增加。3、中耳胆脂瘤组织中VEGF蛋白的平均光密度值为(0.3816±0.04205),正常外耳道皮肤中VEGF蛋白的平均光密度值为(0.2106±0.02105),两组间差异有统计学意义(t=4.804,p<0.01);随着VEGF平均光密度值增加,胆脂瘤上皮基质周围微血管密度(MVD)也相应地增加。4、中耳胆脂瘤组织中MMP9蛋白的平均光密度值为(0.2816±0.04103),正常外耳道皮肤中MMP9蛋白的平均光密度值为(0.2016±0.01803),两组间差异有统计学意义(t=3.184,p<0.01);显著高于正常外耳道皮肤,随着MMP9平均光密度值增加,胆脂瘤上皮基质周围微血管密度(MVD)也相应地增加。5、相关性分析显示VEGF和MMP9在诱导血管生成过程中具有协同作用(r=0.851,p=0.000)。结论:1、中耳胆脂瘤上皮下结缔组织中存在微血管增生。2、胆脂瘤上皮细胞具有较强的增殖活性;胆脂瘤上皮下结缔组织中微血管增生可以为胆脂瘤上皮的异常高度增殖提供较丰富的血供和营养。3、MMP9和VEGF蛋白诱导血管生成在中耳胆脂瘤增殖中起重要作用;MMP9和VEGF在诱导胆脂瘤血管生成过程中具有协同作用。更多还原

【Abstract】 Objective:To observe the expression and significance of VEGF,MMP9,CD105 and Ki-67 in human middle ear cholesteatoma tissues and to investigate the correlation between their expression and the epithelial hyper- proliferation and development of cholesteatoma.The labeling index for Ki-67 was used to assess the proliferative activity and to examine the hyperproliferate state of keratinocyte in cholesteatoma tissues. The CD105 was used to label the microvessel in cholesteatoma tissues.Method:We examined the expression of VEGF,MMP9,CD105 and Ki-67 by immunohistochemical staining in 45 samples from middle ear cholesteatoma and 10 epithelial cells samples from the external auditory meatus. The computer image quantitative analysis was used to examin the expression of VEGF,MMP9, and Ki-67. The microvasculars of cholesteatoma were measured by MVD which were immunohistostained with CD105.Result:1.The microvasculars of cholesteatoma matrix were plentiful and close.But the microvasculars of the skin of external acoustic meatus were less and more raritas than cholesteatoma.The mean MVD was 28.3±7.7(n=45) in cholesteatoma and 6.1±2.5(n=10)in the skin of external acoustic meatus.There were statistical significance between the two groups(t=8.905,p<0.001).2.The the expression of VEGF,MMP9 in cholesteatoma and in normal skin epithelial tissues: The positive immunohistostaining of MMP9 and VEGF appeared all the layers, MMP9 and VEGF protein was localized mostly in the cytoplasm,and it showed the tendency of gradual increasing from basal layer to corneous layer. The normal skin epithelial tissues showed weakly positive.The results showed that VEGF in cholesteatoma tissues (91.43%) was higher than that in normal skin epithelial tissues (40.00%), the expression level of VEGF in cholesteatoma tissues was significantly increased (P<0.01). The results also showed that MMP9 in cholesteatoma tissues (68.57%) was higher than that in normal skin epithelial tissues (20.00%), the expression level of MMP9 in cholesteatoma tissues was significantly increased (P<0.01). The expression of MMP9 and VEGF was present positive correlation in cholesteatoma tissues (P<0.05).3. The results showed that Ki-67 in cholesteatoma tissues (64.44%) was higher than that in normal skin epithelial tissues (20.00%),the expression level of Ki-67 in cholesteatoma tissues was significantly increased(P<0.01). With the increased of mean MVD, the expression level of Ki-67 in cholesteatoma tissues increased.Conclusion:1、there were microvascular proliferation in the presence of cholesteatoma tissues.2、More cells in cell cycle and the strong proliferation activity in cholesteatoma epithelium. Microvascular proliferation can be highly abnormal proliferation of cholesteatoma epithelium provide rich blood supply and nutrition, the high proliferation of epithelial cells in cholesteatoma cholesteatoma matrix dependent on microvascular.3、VEGF and MMP9 protein plays an important role in human middle ear cholesteatoma proliferation. Correlation analysis showed that VEGF and MMP9 in the induction of angiogenesis has a synergistic effect.更多还原