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目的 :研究深低温保存猪二尖瓣内皮细胞结构和功能变化。 方法 :分别取新鲜 (新鲜瓣膜组 )、4℃抗生素灭菌 2 4小时 ( 4℃灭菌 2 4小时组 )以及深低温保存 (深低温保存组 ) 1月猪二尖瓣各 8枚 ,取各自前叶组织 10 mm× 10 mm,M199液中孵育 2 4小时。放射免疫方法测定培养液 6-酮 -前列腺素F1α( 6- keto- PGF1α)浓度。电镜观察深低温保存二尖瓣组织超微结构。 结果 :新鲜瓣膜组、4℃灭菌 2 4小时组以及深低温保存组猪二尖瓣内皮细胞释放 6- keto- PGF1α分别为每平方厘米12 1.5± 16.2 ng/ ml,66.0± 9.5 ng/ ml和 67.3± 6.7ng/ ml。新鲜瓣膜组明显高于 4℃灭菌 2 4小时组和深低温保存组。后两组间无差别。电镜观察到 ,深低温保存二尖瓣内皮细胞与其下的纤维层连接紧密 ,结构完整 ,无损伤性改变。除了胞质中脂褐素类物质较新鲜瓣膜内皮细胞内略增多外 ,与新鲜瓣膜内皮细胞无差异。 结论 :深低温保存瓣膜的内皮细胞结构尚属正常时 ,其功能已经发生损害。深低温保存对内皮细胞的损伤主要发生在 4℃抗生素灭菌过程。优化抗生素组合 ,缩短 4℃灭菌时间 ,将减轻抗生素灭菌过程对内皮细胞的损伤。
Objective: To study the structure and function of mitochondria endothelial cells preserved in cryopreservation pigs. Methods: Fresh mitral valve (fresh valve group), 4 hours antibiotic sterilized at 4 ℃ (24 hours sterilized at 4 ℃), and 8 piglets mitral valve in January at low temperature (cryopreservation group) Each anterior leaf tissue 10 mm × 10 mm, M199 liquid incubated for 24 hours. Radioimmunoassay was used to determine the concentration of 6-keto-PGF1α in culture medium. The ultrastructure of mitral valve was preserved under electron microscope. RESULTS: 6-keto-PGF1α released from mitral valve endothelial cells in fresh valve group, 24 hours sterilized at 4 ℃ and in cryopreservation group were 12 1.5 ± 16.2 ng / ml and 66.0 ± 9.5 ng / ml And 67.3 ± 6.7 ng / ml. Fresh valve group was significantly higher than 4 ℃ sterilization 24 hours group and cryogenic preservation group. After the two groups no difference. Electron microscopy showed that the preservation of mitral endothelial cells in deep hypothermia is tightly connected with the underlying fibrous layer, with structural integrity and no damage changes. In addition to lipofuscin in the cytoplasm of fresh valve endothelial cells slightly increased, with fresh valve endothelial cells no difference. CONCLUSIONS: The endothelial cell structure of the preserved cryopreservation valve is normal and its function has been impaired. Cryopreservation of endothelial cell damage occurs mainly at 4 ℃ antibiotic sterilization process. Optimize the antibiotic combination, shorten the sterilization time of 4 ℃, will reduce the process of antibiotic sterilization of endothelial cell damage.