SOM模型大鼠中耳黏膜及耳积液中TNF-α的表达及抗TNF-α治疗的机制研究

Expression and significance of Transforming Growth Factor-αin the tympanum mucosa of middle ear and venous serum in otitis media with fusion in the guinea pig model

目的探讨抗TNF-α单克隆抗体对大鼠分泌性中耳炎(SOM)的治疗作用及其机制。方法应用随机数字表法将90只健康成年SD大鼠分成三组:对照组、SOM组、抗TNF-α治疗组,每组又根据SOM制成后不同时点,再分为6 h、1 d、3 d、7 d和14 d组,每组6只动物。HE染色后观察中耳黏膜上皮组织病理变化,应用免疫组织化学法检测中耳黏膜上皮组织中TNF-α的表达情况,并采用酶联免疫吸附(ELISA法)检测大鼠耳积液(MEE)中TNF-α的表达水平。结果制模后各时相,对照组标本中无TNF-α的阳性表达,而SOM组和抗TNF-α治疗组中耳黏膜上皮细胞中TNF-α表达灰度值均呈逐渐升高趋势,且较对照组均显著增强(P<0.05)。制模后各时相SOM组和抗TNF-α治疗组之间比较,除6 h组外,抗TNF-α治疗组其余3个时相中耳黏膜上皮细胞中TNF-α表达灰度值均明显低于SOM组(P<0.05)。制模后各时相SOM组MEE中TNF-α的表达水平均高于对照组,呈逐步上升趋势,差异均有显著性(P<0.05)。制模后各时相抗TNF-α治疗组MEE中TNF-α蛋白含量的变化曲线与SOM组相似。两组间不同时间点比较,抗TNF-α治疗组MEE中TNF-α蛋白含量明显均低于SOM组(P<0.05)。结论抗TNF-α治疗通过拮抗TNF-α的炎症介导作用而抑制中耳炎症及免疫反应,有助于减轻急性期炎症反应和阻止慢性SOM的发生。

【Objective】To investigate the effect and pathogenesis of anti-tumor necrosis factor αtreatment on secretory otitis media in rat. 【Methods】90 healthy adult SD rats were randomly divided into 3 groups： control group, SOM group, anti TNF-α treatment group. Each group were divided into group according to different time points（6 h, 1 d, 3 d, 7 d and 14 d after SOM）, 6 rats in each group. To observe the pathological changes of middle ear mucosa tissue by HE staining, expression of TNF-α detection of middle ear mucosa cells by immunohistochemistry method, and to detect rat ear effusion（MEE） expression level of TNF-αby enzyme linked immunosorbent assay（ELISA method）. 【Results】The expression of TNF-α in middle ear mucosa cells of control group was. The expression of TNF-α in SOM group and anti TNF-α treatment group were significantly higher than that of control group（P〈0.05）. However, except for 6 h group, at the remaining 3 phase the expression of TNF- α in anti TNF- αtreatment group were significantly lower than those of SOM group（P〈0.05）. At each phase, the level of TNF-α in MEE of SOM group was higher than that of control group, showed a gradual upward trend, the differences were statistically significant（P〈0.05）. The level of TNF-α in MEE of anti TNF-α treatment group was similar to that of SOM group, Comparing the two groups at different time points, the level of TNF-α in the MEE of anti TNF-αtreatment group was significantly lower than that of SOM group（P〈0.05）. 【Conclusions】Anti TNF-α therapy guided by inflammation of antagonistic TNF-α and inhibit ear inflammation and immune reaction, which helps reduce the inflammatory response in acute phase and prevent SOM.