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目的:建立转基因小鼠主动脉瓣疾病模型的评估标准。方法:采用3月龄C57成年小鼠20只,分离小鼠心脏称重后固定,经HE染色、免疫荧光、激光共聚焦和Massoon染色后光镜观察及电镜观察,分别对瓣膜的细胞数量、分布、胶原含量及瓣膜超微结构进行组织形态学分析。结果:正常小鼠主动脉瓣的面积为(0.1978±0.003)mm2,周长为(9.18±0.06)×102μm,平均细胞数为(9.75±0.04)×102个/mm2,其中内皮细胞数为(3.23±0.10)×102个/mm2,间质细胞数为(5.05±0.07)×102个/mm2,胶原含量为(23.77±8.38)%。此外,通过激光共聚焦显微镜观察还发现,成体内皮下层存在一些共表达CD31及α-SMA的细胞,表明成体瓣膜仍保留具有上皮-间质转化潜能的细胞。电镜观察发现,瓣膜的心室面内皮与主动脉面内皮在形态上存在有很大的差异:心室面内皮呈扁平形与血流方向一致,主动脉面内皮呈方形与血流方向垂直。间质中成纤维细胞处于静止状态,其细胞胞体较小,呈长梭形,粗面内质网和高尔基复合体均不发达。细胞外基质胶原的含量丰富,主要集中于流出道。结论:本实验首次详细观测了小鼠主动脉瓣形态学特征,建立了正常小鼠主动脉瓣组织形态学研究的标准,为今后小鼠主动脉瓣疾病模型的研究提供重要的参考依据。
Objective: To establish the evaluation criteria of aortic valve disease model in transgenic mice. Methods: Twenty adult C57 mice aged 3 months were weighed, fixed and fixed in the heart of the isolated mice. The number of the heart, the number of the valve, the diameter of the valve and the diameter of the valve were measured by HE staining, immunofluorescence, confocal laser scanning microscopy and Masson staining, Distribution, collagen content and valvular ultrastructure histomorphology analysis. Results: The area of aortic valve in normal mice was (0.1978 ± 0.003) mm2, the circumference was (9.18 ± 0.06) × 102μm, the average number of cells was (9.75 ± 0.04) × 102 cells / mm2, 3.23 ± 0.10) × 102 cells / mm2, the number of stromal cells was (5.05 ± 0.07) × 102 cells / mm2, and the collagen content was (23.77 ± 8.38)%. In addition, it was also observed by laser confocal microscopy that some cells co-expressing CD31 and α-SMA existed in the subendothelial subtype, indicating that the adult valve retains cells with epithelial-mesenchymal transition potential. Electron microscopy showed that there was a great difference in morphology between the ventricular endothelium of the valve and the aortic endothelium: the flat surface of the ventricular surface was in the same direction as the blood flow, and the aortic endothelium was perpendicular to the direction of the blood flow. Interstitial fibroblasts in a quiescent state, the cell body is small, long fusiform, rough endoplasmic reticulum and Golgi complex are underdeveloped. Extracellular matrix collagen content is rich, mainly in the outflow tract. CONCLUSIONS: For the first time, morphological characteristics of mouse aortic valve were observed in detail and the standard of aortic valve tissue morphology was established, which provided important reference for the study of mouse aortic valve disease model in the future.