局部应用钙拮抗剂抑制鼓室硬化豚鼠模型的初步报告

发布时间：2018-03-28 19:39

本文选题：鼓室硬化　切入点：动物模型　出处：《中华耳科学杂志》2014年02期

【摘要】：目的通过豚鼠中耳炎模型探讨局部应用钙离子拮抗剂抑制鼓室硬化的可能机制。方法通过对豚鼠人工创伤双中耳种植病原菌的方式建立慢性中耳炎的模型,并对左耳每日滴用钙离子拮抗剂地尔硫卓(CBBS)液灌注治疗,右耳为对照耳,滴生理盐水的方法,来研究中耳局部使用钙离子拮抗剂是否能通过抑制巨噬细胞的趋化分泌活动,进而抑制骨形成蛋白-2表达,较少钙盐沉积,从而阻止或延缓MS、TS发生。观察数据1)光学显微镜下观察鼓室粘膜炎性细胞、肉芽组织及硬化灶病理学改变;2)分别使用CD68及BMP-2抗体进行标记,确定中耳组织内侵入的细胞类型和BMP-2表达情况。3)取中耳近鼓膜处标本送投射电镜观察粘膜组织纤维细胞内形态改变、巨嗜细胞趋化情况及组织内纤维增生和钙离子沉着情况。结果光学显微镜下对照耳听泡内炎症较实验耳严重;CD68及BMP-2染色情况:实验耳及对照耳粘膜及听泡内均有较高CD46的表达并和感染的严重度呈正比,对照耳稍多;对照耳粘膜BMP-2染色明显高于实验耳,即使感染程度接近仍有明显差异,并且在粘膜下层会出现较密集的群聚现象,考虑为大量的巨嗜细胞集聚并有较高的BMP-2分泌。电镜改变:实验耳标本内尚可见部分柱状上皮细胞,仅有少量胶原纤维增生,炎症细胞淋巴细胞为主,很少见到巨嗜细胞,基本看不到钙盐沉积。对照耳标本组织破坏严重,部分区域几乎看不到多少正常组织细胞,已无法辨认出柱状上皮细胞,可见成堆的充满吞噬泡的巨嗜细胞,胶原纤维增生明显,并伴有成堆的钙盐沉积。结论动物模型实验提示中耳局部使用钙离子拮抗剂能通过抑制巨噬细胞的趋化分泌活动,减轻局部炎症破坏,进而抑制骨形成蛋白-2表达,较少钙盐沉积,从而阻止或延缓鼓室硬化的发生。
[Abstract]:Objective to explore the possible mechanism of local application of calcium antagonist in inhibiting tympanic sclerosis in guinea pig otitis media. The left ear was treated with calcium antagonist diltiazem (CBBSs) daily, and the right ear was used as control ear, and normal saline was used to study whether local use of calcium antagonist in middle ear could inhibit the chemotaxis of macrophages. In turn, the expression of bone morphogenetic protein -2 was inhibited and the formation of MSTS was prevented or delayed by less calcium deposition. Observation data 1) the inflammatory cells of tympanic mucosa were observed under optical microscope. CD68 and BMP-2 antibodies were used to label granulation tissues and sclerosing foci respectively. To determine the type of invading cells and the expression of BMP-2 in the middle ear tissue. 3) the specimens of the middle ear near the tympanic membrane were taken to project electron microscope to observe the morphological changes of the mucosal fibroblasts. Results under optical microscope, inflammation in auditory vesicles was more serious than that in experimental ears, CD68 and BMP-2 staining: the mucous membranes and auditory vesicles of experimental and control ears were found in the experimental and control ears. The high expression of CD46 was proportional to the severity of infection. The mucosal BMP-2 staining of the control ear was significantly higher than that of the experimental ear, even though the infection degree was close to that of the experimental ear, there were significant differences, and there was a dense clustering phenomenon in the submucous membrane. Electron microscopic changes: some columnar epithelial cells were found in the ear specimens, only a small number of collagen fibers proliferated, inflammatory cells were mainly lymphocytes, and few giant eosinophil cells were found. There was no calcium deposition. In contrast, the tissue in the ear label was severely damaged, and few normal tissue cells were found in some areas. The columnar epithelial cells could no longer be recognized, and piles of giant eosinophils filled with phagocytic bubbles could be seen. Conclusion the experimental results suggest that calcium antagonist can reduce the local inflammation by inhibiting the chemotaxis and secretion of macrophages. Then, the expression of bone morphogenetic protein-2 was inhibited, and the formation of tympanic sclerosis was prevented or delayed.
【作者单位】：上海交通大学附属第一人民医院分院耳鼻喉科;
【基金】：虹卫1102-01(上海市虹口区),局部应用钙拮抗剂抑制鼓膜鼓室硬化的动物模型实验和临床研究
【分类号】：R764.2

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